Illana Gozes PhD Professor of Clinical Biochemistry Lily and Avraham Gildor Chair for the Investigation of Growth Factors Adams Super Center for Brain Studies amp Edersheim Levie-Gitter fMRI Institute (wwwbraintauacil) Head the Dr Diana and Zelman Elton (Elbaum) Laboratory for Molecular Neuroendocrinology Department of Human Molecular Genetics and Biochemistry Tel Aviv University Sackler Faculty of Medicine Sagol School of Neuroscience Israel
Laboratory Members PhD students Anna Malishkevich Yanina Ivashco-Pachima Avia Merenlender-Wagner Adva Hadar Iris Grigg and Shlomi Sragovich Master Students Evgenia Maryanovsky Regina Ostritsky Sinaya Vaisburd Gal Hacohen-Kleiman Noy Amram Livnat Bacal and Yarden Ziv Dr Eliezer Giladi PhD Laboratory Manager
TAU collaborators Dr David Gurwitz Dr Segev Barak Dr Rina Rosin-Arbesfeld Dr Joel Hirsch Dr Leonid Mittelman Dr Metsada Pasmanik-Chor and Adva Yeheskel
Molecules that make our minds fighting autism schizophrenia and Alzheimerrsquos disease
Imagine your brain as a large rail transportation system and your 100 billion nerve cells = trains while your brain ldquosupport cellsrdquo are the train tracks
Now imagine disruption in connectivity hellip (~100 trillion points of connection)hellip
Can we fix it
And he does not talk
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
Cognitive Development and Aging Dementia
Autism Intellectual disability and facial dysmorphisms caused by mutations in
ADNP Nature Genetics 46 380ndash384 (2014)
Nature Genetics
Volume
46
Pages
380ndash384
Year published
(2014)
DOI
httpwwwncbinlmnihgovgtrtests515961methodology
And he does not talk
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
Cognitive Development and Aging Dementia
Autism Intellectual disability and facial dysmorphisms caused by mutations in
ADNP Nature Genetics 46 380ndash384 (2014)
Nature Genetics
Volume
46
Pages
380ndash384
Year published
(2014)
DOI
httpwwwncbinlmnihgovgtrtests515961methodology
Autism Spectrum Disorder
Autism spectrum disorder (ASD) refers to a range of disorders lsquotriad of impairmentsrsquo
(1) social and emotional understanding
(2) all aspects of communication
(3) flexibility in thinking and behavior
(Wing 1996)
Dr Hans Asperger Dr Leo Kanner
Connecting Autism Cancer and Alzheimerrsquos disease
TauAutophagy Apoptosis Regulating gt400 Genes
ADNP
Disease Injury
Brain support cell-derived protectants
Our Discovery Protective Platforms
ADNP platform
Davunetide NAP D-NAP
DementiaCognitive impairment
Alzheimerrsquos and Parkinsonrsquos disease schizophrenia
ADNF platform
D-SAL AL-309 (9 aa)
SAL AL-209 (9 aa)
Neuropathies
Activity-Dependent Neurotrophic Factor
Activity-Dependent Neuroprotective Protein
NAPVSIPQ SALLRSIPA
Brenneman DE Gozes I (1996) A femtomolar-acting neuroprotective peptide J Clin Invest 972299-2307
Bassan M Zamostiano R Davidson A Pinhasov A Giladi E Perl O Bassan H Blat C Gibney G Glazner G Brenneman DE Gozes I (1999) Complete sequence of a novel protein containing a femtomolar-activity-dependent neuroprotective peptide J Neurochem 721283-1293
Neuropeptides
Activity-Dependent Neuroprotective Protein (ADNP)
Our papers J Biol Chem 2012 287(48)40173-85 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 J Pharmacol Exp Ther 2007323438-434 Pharmacol Ther 2007 114146-54 CNS Drug Rev 200511353-68 Brain Res Dev Brain Res 200314483-90 Neuron Glia Biol 2004 Aug1(3)193-199
gt 1 No ADNP ndash No brain
gt 2 ADNP is part of a large protein complex - SWISNF regulating gt400 genes and determining nerve cell shape and brain structure
gt 3 As part of the SWISNF ADNP associated with
proteins regulating tau splicing
1
2
2
Schirer Y Malishkevich A Ophir Y Lewis J Giladi E Gozes I Novel Marker for the Onset of Frontotemporal Dementia Early Increase in Activity-Dependent Neuroprotective Protein (ADNP) in the Face of Tau Mutation PLoS One 2014 9(1) e87383
3
Pinhasov A Mandel S Torchinsky A Giladi E Pittel Z Goldsweig AM Servoss SJ Brenneman DE Gozes I (2003) Activity-dependent neuroprotective protein a novel gene essential for brain formation Brain Res Dev Brain Res 144 83-90
Organogenesis
Neuroogenesis
Regulation of
Transcription
Lytic
vacuole
Lipid
Transport
Coagulation
E9-95 ADNP knock out embryo
ADNP regulates ApoE ndash the major risk gene for Alzheimerrsquos disease
Gene Array ADNP Regulates gt400 Genes
Mandel S Rechavi G Gozes I (2007) Activity-dependent neuroprotective protein (ADNP) differentially interacts with chromatin to regulate genes essential for embryogenesis Dev Biol 303 814-824
Bioinformatics The ADNP Network
Gozes Yeheskel Pasmanik-Chor Activity-Dependent Neuroprotective Protein (ADNP) A Case Study for Highly Conserved Chordata-Specific Genes Shaping the Brain and Mutated in Cancer J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
httpwwwadnpgenecom Feb (0) doi 101038ng2899 Nat Genet 46380-384
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Imagine your brain as a large rail transportation system and your 100 billion nerve cells = trains while your brain ldquosupport cellsrdquo are the train tracks
Now imagine disruption in connectivity hellip (~100 trillion points of connection)hellip
Can we fix it
And he does not talk
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
Cognitive Development and Aging Dementia
Autism Intellectual disability and facial dysmorphisms caused by mutations in
ADNP Nature Genetics 46 380ndash384 (2014)
Nature Genetics
Volume
46
Pages
380ndash384
Year published
(2014)
DOI
httpwwwncbinlmnihgovgtrtests515961methodology
And he does not talk
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
Cognitive Development and Aging Dementia
Autism Intellectual disability and facial dysmorphisms caused by mutations in
ADNP Nature Genetics 46 380ndash384 (2014)
Nature Genetics
Volume
46
Pages
380ndash384
Year published
(2014)
DOI
httpwwwncbinlmnihgovgtrtests515961methodology
Autism Spectrum Disorder
Autism spectrum disorder (ASD) refers to a range of disorders lsquotriad of impairmentsrsquo
(1) social and emotional understanding
(2) all aspects of communication
(3) flexibility in thinking and behavior
(Wing 1996)
Dr Hans Asperger Dr Leo Kanner
Connecting Autism Cancer and Alzheimerrsquos disease
TauAutophagy Apoptosis Regulating gt400 Genes
ADNP
Disease Injury
Brain support cell-derived protectants
Our Discovery Protective Platforms
ADNP platform
Davunetide NAP D-NAP
DementiaCognitive impairment
Alzheimerrsquos and Parkinsonrsquos disease schizophrenia
ADNF platform
D-SAL AL-309 (9 aa)
SAL AL-209 (9 aa)
Neuropathies
Activity-Dependent Neurotrophic Factor
Activity-Dependent Neuroprotective Protein
NAPVSIPQ SALLRSIPA
Brenneman DE Gozes I (1996) A femtomolar-acting neuroprotective peptide J Clin Invest 972299-2307
Bassan M Zamostiano R Davidson A Pinhasov A Giladi E Perl O Bassan H Blat C Gibney G Glazner G Brenneman DE Gozes I (1999) Complete sequence of a novel protein containing a femtomolar-activity-dependent neuroprotective peptide J Neurochem 721283-1293
Neuropeptides
Activity-Dependent Neuroprotective Protein (ADNP)
Our papers J Biol Chem 2012 287(48)40173-85 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 J Pharmacol Exp Ther 2007323438-434 Pharmacol Ther 2007 114146-54 CNS Drug Rev 200511353-68 Brain Res Dev Brain Res 200314483-90 Neuron Glia Biol 2004 Aug1(3)193-199
gt 1 No ADNP ndash No brain
gt 2 ADNP is part of a large protein complex - SWISNF regulating gt400 genes and determining nerve cell shape and brain structure
gt 3 As part of the SWISNF ADNP associated with
proteins regulating tau splicing
1
2
2
Schirer Y Malishkevich A Ophir Y Lewis J Giladi E Gozes I Novel Marker for the Onset of Frontotemporal Dementia Early Increase in Activity-Dependent Neuroprotective Protein (ADNP) in the Face of Tau Mutation PLoS One 2014 9(1) e87383
3
Pinhasov A Mandel S Torchinsky A Giladi E Pittel Z Goldsweig AM Servoss SJ Brenneman DE Gozes I (2003) Activity-dependent neuroprotective protein a novel gene essential for brain formation Brain Res Dev Brain Res 144 83-90
Organogenesis
Neuroogenesis
Regulation of
Transcription
Lytic
vacuole
Lipid
Transport
Coagulation
E9-95 ADNP knock out embryo
ADNP regulates ApoE ndash the major risk gene for Alzheimerrsquos disease
Gene Array ADNP Regulates gt400 Genes
Mandel S Rechavi G Gozes I (2007) Activity-dependent neuroprotective protein (ADNP) differentially interacts with chromatin to regulate genes essential for embryogenesis Dev Biol 303 814-824
Bioinformatics The ADNP Network
Gozes Yeheskel Pasmanik-Chor Activity-Dependent Neuroprotective Protein (ADNP) A Case Study for Highly Conserved Chordata-Specific Genes Shaping the Brain and Mutated in Cancer J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
httpwwwadnpgenecom Feb (0) doi 101038ng2899 Nat Genet 46380-384
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
And he does not talk
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
Cognitive Development and Aging Dementia
Autism Intellectual disability and facial dysmorphisms caused by mutations in
ADNP Nature Genetics 46 380ndash384 (2014)
Nature Genetics
Volume
46
Pages
380ndash384
Year published
(2014)
DOI
httpwwwncbinlmnihgovgtrtests515961methodology
And he does not talk
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
Cognitive Development and Aging Dementia
Autism Intellectual disability and facial dysmorphisms caused by mutations in
ADNP Nature Genetics 46 380ndash384 (2014)
Nature Genetics
Volume
46
Pages
380ndash384
Year published
(2014)
DOI
httpwwwncbinlmnihgovgtrtests515961methodology
Autism Spectrum Disorder
Autism spectrum disorder (ASD) refers to a range of disorders lsquotriad of impairmentsrsquo
(1) social and emotional understanding
(2) all aspects of communication
(3) flexibility in thinking and behavior
(Wing 1996)
Dr Hans Asperger Dr Leo Kanner
Connecting Autism Cancer and Alzheimerrsquos disease
TauAutophagy Apoptosis Regulating gt400 Genes
ADNP
Disease Injury
Brain support cell-derived protectants
Our Discovery Protective Platforms
ADNP platform
Davunetide NAP D-NAP
DementiaCognitive impairment
Alzheimerrsquos and Parkinsonrsquos disease schizophrenia
ADNF platform
D-SAL AL-309 (9 aa)
SAL AL-209 (9 aa)
Neuropathies
Activity-Dependent Neurotrophic Factor
Activity-Dependent Neuroprotective Protein
NAPVSIPQ SALLRSIPA
Brenneman DE Gozes I (1996) A femtomolar-acting neuroprotective peptide J Clin Invest 972299-2307
Bassan M Zamostiano R Davidson A Pinhasov A Giladi E Perl O Bassan H Blat C Gibney G Glazner G Brenneman DE Gozes I (1999) Complete sequence of a novel protein containing a femtomolar-activity-dependent neuroprotective peptide J Neurochem 721283-1293
Neuropeptides
Activity-Dependent Neuroprotective Protein (ADNP)
Our papers J Biol Chem 2012 287(48)40173-85 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 J Pharmacol Exp Ther 2007323438-434 Pharmacol Ther 2007 114146-54 CNS Drug Rev 200511353-68 Brain Res Dev Brain Res 200314483-90 Neuron Glia Biol 2004 Aug1(3)193-199
gt 1 No ADNP ndash No brain
gt 2 ADNP is part of a large protein complex - SWISNF regulating gt400 genes and determining nerve cell shape and brain structure
gt 3 As part of the SWISNF ADNP associated with
proteins regulating tau splicing
1
2
2
Schirer Y Malishkevich A Ophir Y Lewis J Giladi E Gozes I Novel Marker for the Onset of Frontotemporal Dementia Early Increase in Activity-Dependent Neuroprotective Protein (ADNP) in the Face of Tau Mutation PLoS One 2014 9(1) e87383
3
Pinhasov A Mandel S Torchinsky A Giladi E Pittel Z Goldsweig AM Servoss SJ Brenneman DE Gozes I (2003) Activity-dependent neuroprotective protein a novel gene essential for brain formation Brain Res Dev Brain Res 144 83-90
Organogenesis
Neuroogenesis
Regulation of
Transcription
Lytic
vacuole
Lipid
Transport
Coagulation
E9-95 ADNP knock out embryo
ADNP regulates ApoE ndash the major risk gene for Alzheimerrsquos disease
Gene Array ADNP Regulates gt400 Genes
Mandel S Rechavi G Gozes I (2007) Activity-dependent neuroprotective protein (ADNP) differentially interacts with chromatin to regulate genes essential for embryogenesis Dev Biol 303 814-824
Bioinformatics The ADNP Network
Gozes Yeheskel Pasmanik-Chor Activity-Dependent Neuroprotective Protein (ADNP) A Case Study for Highly Conserved Chordata-Specific Genes Shaping the Brain and Mutated in Cancer J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
httpwwwadnpgenecom Feb (0) doi 101038ng2899 Nat Genet 46380-384
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
And he does not talk
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
Cognitive Development and Aging Dementia
Autism Intellectual disability and facial dysmorphisms caused by mutations in
ADNP Nature Genetics 46 380ndash384 (2014)
Nature Genetics
Volume
46
Pages
380ndash384
Year published
(2014)
DOI
httpwwwncbinlmnihgovgtrtests515961methodology
Autism Spectrum Disorder
Autism spectrum disorder (ASD) refers to a range of disorders lsquotriad of impairmentsrsquo
(1) social and emotional understanding
(2) all aspects of communication
(3) flexibility in thinking and behavior
(Wing 1996)
Dr Hans Asperger Dr Leo Kanner
Connecting Autism Cancer and Alzheimerrsquos disease
TauAutophagy Apoptosis Regulating gt400 Genes
ADNP
Disease Injury
Brain support cell-derived protectants
Our Discovery Protective Platforms
ADNP platform
Davunetide NAP D-NAP
DementiaCognitive impairment
Alzheimerrsquos and Parkinsonrsquos disease schizophrenia
ADNF platform
D-SAL AL-309 (9 aa)
SAL AL-209 (9 aa)
Neuropathies
Activity-Dependent Neurotrophic Factor
Activity-Dependent Neuroprotective Protein
NAPVSIPQ SALLRSIPA
Brenneman DE Gozes I (1996) A femtomolar-acting neuroprotective peptide J Clin Invest 972299-2307
Bassan M Zamostiano R Davidson A Pinhasov A Giladi E Perl O Bassan H Blat C Gibney G Glazner G Brenneman DE Gozes I (1999) Complete sequence of a novel protein containing a femtomolar-activity-dependent neuroprotective peptide J Neurochem 721283-1293
Neuropeptides
Activity-Dependent Neuroprotective Protein (ADNP)
Our papers J Biol Chem 2012 287(48)40173-85 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 J Pharmacol Exp Ther 2007323438-434 Pharmacol Ther 2007 114146-54 CNS Drug Rev 200511353-68 Brain Res Dev Brain Res 200314483-90 Neuron Glia Biol 2004 Aug1(3)193-199
gt 1 No ADNP ndash No brain
gt 2 ADNP is part of a large protein complex - SWISNF regulating gt400 genes and determining nerve cell shape and brain structure
gt 3 As part of the SWISNF ADNP associated with
proteins regulating tau splicing
1
2
2
Schirer Y Malishkevich A Ophir Y Lewis J Giladi E Gozes I Novel Marker for the Onset of Frontotemporal Dementia Early Increase in Activity-Dependent Neuroprotective Protein (ADNP) in the Face of Tau Mutation PLoS One 2014 9(1) e87383
3
Pinhasov A Mandel S Torchinsky A Giladi E Pittel Z Goldsweig AM Servoss SJ Brenneman DE Gozes I (2003) Activity-dependent neuroprotective protein a novel gene essential for brain formation Brain Res Dev Brain Res 144 83-90
Organogenesis
Neuroogenesis
Regulation of
Transcription
Lytic
vacuole
Lipid
Transport
Coagulation
E9-95 ADNP knock out embryo
ADNP regulates ApoE ndash the major risk gene for Alzheimerrsquos disease
Gene Array ADNP Regulates gt400 Genes
Mandel S Rechavi G Gozes I (2007) Activity-dependent neuroprotective protein (ADNP) differentially interacts with chromatin to regulate genes essential for embryogenesis Dev Biol 303 814-824
Bioinformatics The ADNP Network
Gozes Yeheskel Pasmanik-Chor Activity-Dependent Neuroprotective Protein (ADNP) A Case Study for Highly Conserved Chordata-Specific Genes Shaping the Brain and Mutated in Cancer J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
httpwwwadnpgenecom Feb (0) doi 101038ng2899 Nat Genet 46380-384
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Autism Spectrum Disorder
Autism spectrum disorder (ASD) refers to a range of disorders lsquotriad of impairmentsrsquo
(1) social and emotional understanding
(2) all aspects of communication
(3) flexibility in thinking and behavior
(Wing 1996)
Dr Hans Asperger Dr Leo Kanner
Connecting Autism Cancer and Alzheimerrsquos disease
TauAutophagy Apoptosis Regulating gt400 Genes
ADNP
Disease Injury
Brain support cell-derived protectants
Our Discovery Protective Platforms
ADNP platform
Davunetide NAP D-NAP
DementiaCognitive impairment
Alzheimerrsquos and Parkinsonrsquos disease schizophrenia
ADNF platform
D-SAL AL-309 (9 aa)
SAL AL-209 (9 aa)
Neuropathies
Activity-Dependent Neurotrophic Factor
Activity-Dependent Neuroprotective Protein
NAPVSIPQ SALLRSIPA
Brenneman DE Gozes I (1996) A femtomolar-acting neuroprotective peptide J Clin Invest 972299-2307
Bassan M Zamostiano R Davidson A Pinhasov A Giladi E Perl O Bassan H Blat C Gibney G Glazner G Brenneman DE Gozes I (1999) Complete sequence of a novel protein containing a femtomolar-activity-dependent neuroprotective peptide J Neurochem 721283-1293
Neuropeptides
Activity-Dependent Neuroprotective Protein (ADNP)
Our papers J Biol Chem 2012 287(48)40173-85 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 J Pharmacol Exp Ther 2007323438-434 Pharmacol Ther 2007 114146-54 CNS Drug Rev 200511353-68 Brain Res Dev Brain Res 200314483-90 Neuron Glia Biol 2004 Aug1(3)193-199
gt 1 No ADNP ndash No brain
gt 2 ADNP is part of a large protein complex - SWISNF regulating gt400 genes and determining nerve cell shape and brain structure
gt 3 As part of the SWISNF ADNP associated with
proteins regulating tau splicing
1
2
2
Schirer Y Malishkevich A Ophir Y Lewis J Giladi E Gozes I Novel Marker for the Onset of Frontotemporal Dementia Early Increase in Activity-Dependent Neuroprotective Protein (ADNP) in the Face of Tau Mutation PLoS One 2014 9(1) e87383
3
Pinhasov A Mandel S Torchinsky A Giladi E Pittel Z Goldsweig AM Servoss SJ Brenneman DE Gozes I (2003) Activity-dependent neuroprotective protein a novel gene essential for brain formation Brain Res Dev Brain Res 144 83-90
Organogenesis
Neuroogenesis
Regulation of
Transcription
Lytic
vacuole
Lipid
Transport
Coagulation
E9-95 ADNP knock out embryo
ADNP regulates ApoE ndash the major risk gene for Alzheimerrsquos disease
Gene Array ADNP Regulates gt400 Genes
Mandel S Rechavi G Gozes I (2007) Activity-dependent neuroprotective protein (ADNP) differentially interacts with chromatin to regulate genes essential for embryogenesis Dev Biol 303 814-824
Bioinformatics The ADNP Network
Gozes Yeheskel Pasmanik-Chor Activity-Dependent Neuroprotective Protein (ADNP) A Case Study for Highly Conserved Chordata-Specific Genes Shaping the Brain and Mutated in Cancer J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
httpwwwadnpgenecom Feb (0) doi 101038ng2899 Nat Genet 46380-384
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Connecting Autism Cancer and Alzheimerrsquos disease
TauAutophagy Apoptosis Regulating gt400 Genes
ADNP
Disease Injury
Brain support cell-derived protectants
Our Discovery Protective Platforms
ADNP platform
Davunetide NAP D-NAP
DementiaCognitive impairment
Alzheimerrsquos and Parkinsonrsquos disease schizophrenia
ADNF platform
D-SAL AL-309 (9 aa)
SAL AL-209 (9 aa)
Neuropathies
Activity-Dependent Neurotrophic Factor
Activity-Dependent Neuroprotective Protein
NAPVSIPQ SALLRSIPA
Brenneman DE Gozes I (1996) A femtomolar-acting neuroprotective peptide J Clin Invest 972299-2307
Bassan M Zamostiano R Davidson A Pinhasov A Giladi E Perl O Bassan H Blat C Gibney G Glazner G Brenneman DE Gozes I (1999) Complete sequence of a novel protein containing a femtomolar-activity-dependent neuroprotective peptide J Neurochem 721283-1293
Neuropeptides
Activity-Dependent Neuroprotective Protein (ADNP)
Our papers J Biol Chem 2012 287(48)40173-85 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 J Pharmacol Exp Ther 2007323438-434 Pharmacol Ther 2007 114146-54 CNS Drug Rev 200511353-68 Brain Res Dev Brain Res 200314483-90 Neuron Glia Biol 2004 Aug1(3)193-199
gt 1 No ADNP ndash No brain
gt 2 ADNP is part of a large protein complex - SWISNF regulating gt400 genes and determining nerve cell shape and brain structure
gt 3 As part of the SWISNF ADNP associated with
proteins regulating tau splicing
1
2
2
Schirer Y Malishkevich A Ophir Y Lewis J Giladi E Gozes I Novel Marker for the Onset of Frontotemporal Dementia Early Increase in Activity-Dependent Neuroprotective Protein (ADNP) in the Face of Tau Mutation PLoS One 2014 9(1) e87383
3
Pinhasov A Mandel S Torchinsky A Giladi E Pittel Z Goldsweig AM Servoss SJ Brenneman DE Gozes I (2003) Activity-dependent neuroprotective protein a novel gene essential for brain formation Brain Res Dev Brain Res 144 83-90
Organogenesis
Neuroogenesis
Regulation of
Transcription
Lytic
vacuole
Lipid
Transport
Coagulation
E9-95 ADNP knock out embryo
ADNP regulates ApoE ndash the major risk gene for Alzheimerrsquos disease
Gene Array ADNP Regulates gt400 Genes
Mandel S Rechavi G Gozes I (2007) Activity-dependent neuroprotective protein (ADNP) differentially interacts with chromatin to regulate genes essential for embryogenesis Dev Biol 303 814-824
Bioinformatics The ADNP Network
Gozes Yeheskel Pasmanik-Chor Activity-Dependent Neuroprotective Protein (ADNP) A Case Study for Highly Conserved Chordata-Specific Genes Shaping the Brain and Mutated in Cancer J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
httpwwwadnpgenecom Feb (0) doi 101038ng2899 Nat Genet 46380-384
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Disease Injury
Brain support cell-derived protectants
Our Discovery Protective Platforms
ADNP platform
Davunetide NAP D-NAP
DementiaCognitive impairment
Alzheimerrsquos and Parkinsonrsquos disease schizophrenia
ADNF platform
D-SAL AL-309 (9 aa)
SAL AL-209 (9 aa)
Neuropathies
Activity-Dependent Neurotrophic Factor
Activity-Dependent Neuroprotective Protein
NAPVSIPQ SALLRSIPA
Brenneman DE Gozes I (1996) A femtomolar-acting neuroprotective peptide J Clin Invest 972299-2307
Bassan M Zamostiano R Davidson A Pinhasov A Giladi E Perl O Bassan H Blat C Gibney G Glazner G Brenneman DE Gozes I (1999) Complete sequence of a novel protein containing a femtomolar-activity-dependent neuroprotective peptide J Neurochem 721283-1293
Neuropeptides
Activity-Dependent Neuroprotective Protein (ADNP)
Our papers J Biol Chem 2012 287(48)40173-85 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 J Pharmacol Exp Ther 2007323438-434 Pharmacol Ther 2007 114146-54 CNS Drug Rev 200511353-68 Brain Res Dev Brain Res 200314483-90 Neuron Glia Biol 2004 Aug1(3)193-199
gt 1 No ADNP ndash No brain
gt 2 ADNP is part of a large protein complex - SWISNF regulating gt400 genes and determining nerve cell shape and brain structure
gt 3 As part of the SWISNF ADNP associated with
proteins regulating tau splicing
1
2
2
Schirer Y Malishkevich A Ophir Y Lewis J Giladi E Gozes I Novel Marker for the Onset of Frontotemporal Dementia Early Increase in Activity-Dependent Neuroprotective Protein (ADNP) in the Face of Tau Mutation PLoS One 2014 9(1) e87383
3
Pinhasov A Mandel S Torchinsky A Giladi E Pittel Z Goldsweig AM Servoss SJ Brenneman DE Gozes I (2003) Activity-dependent neuroprotective protein a novel gene essential for brain formation Brain Res Dev Brain Res 144 83-90
Organogenesis
Neuroogenesis
Regulation of
Transcription
Lytic
vacuole
Lipid
Transport
Coagulation
E9-95 ADNP knock out embryo
ADNP regulates ApoE ndash the major risk gene for Alzheimerrsquos disease
Gene Array ADNP Regulates gt400 Genes
Mandel S Rechavi G Gozes I (2007) Activity-dependent neuroprotective protein (ADNP) differentially interacts with chromatin to regulate genes essential for embryogenesis Dev Biol 303 814-824
Bioinformatics The ADNP Network
Gozes Yeheskel Pasmanik-Chor Activity-Dependent Neuroprotective Protein (ADNP) A Case Study for Highly Conserved Chordata-Specific Genes Shaping the Brain and Mutated in Cancer J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
httpwwwadnpgenecom Feb (0) doi 101038ng2899 Nat Genet 46380-384
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Activity-Dependent Neuroprotective Protein (ADNP)
Our papers J Biol Chem 2012 287(48)40173-85 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 J Pharmacol Exp Ther 2007323438-434 Pharmacol Ther 2007 114146-54 CNS Drug Rev 200511353-68 Brain Res Dev Brain Res 200314483-90 Neuron Glia Biol 2004 Aug1(3)193-199
gt 1 No ADNP ndash No brain
gt 2 ADNP is part of a large protein complex - SWISNF regulating gt400 genes and determining nerve cell shape and brain structure
gt 3 As part of the SWISNF ADNP associated with
proteins regulating tau splicing
1
2
2
Schirer Y Malishkevich A Ophir Y Lewis J Giladi E Gozes I Novel Marker for the Onset of Frontotemporal Dementia Early Increase in Activity-Dependent Neuroprotective Protein (ADNP) in the Face of Tau Mutation PLoS One 2014 9(1) e87383
3
Pinhasov A Mandel S Torchinsky A Giladi E Pittel Z Goldsweig AM Servoss SJ Brenneman DE Gozes I (2003) Activity-dependent neuroprotective protein a novel gene essential for brain formation Brain Res Dev Brain Res 144 83-90
Organogenesis
Neuroogenesis
Regulation of
Transcription
Lytic
vacuole
Lipid
Transport
Coagulation
E9-95 ADNP knock out embryo
ADNP regulates ApoE ndash the major risk gene for Alzheimerrsquos disease
Gene Array ADNP Regulates gt400 Genes
Mandel S Rechavi G Gozes I (2007) Activity-dependent neuroprotective protein (ADNP) differentially interacts with chromatin to regulate genes essential for embryogenesis Dev Biol 303 814-824
Bioinformatics The ADNP Network
Gozes Yeheskel Pasmanik-Chor Activity-Dependent Neuroprotective Protein (ADNP) A Case Study for Highly Conserved Chordata-Specific Genes Shaping the Brain and Mutated in Cancer J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
httpwwwadnpgenecom Feb (0) doi 101038ng2899 Nat Genet 46380-384
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Organogenesis
Neuroogenesis
Regulation of
Transcription
Lytic
vacuole
Lipid
Transport
Coagulation
E9-95 ADNP knock out embryo
ADNP regulates ApoE ndash the major risk gene for Alzheimerrsquos disease
Gene Array ADNP Regulates gt400 Genes
Mandel S Rechavi G Gozes I (2007) Activity-dependent neuroprotective protein (ADNP) differentially interacts with chromatin to regulate genes essential for embryogenesis Dev Biol 303 814-824
Bioinformatics The ADNP Network
Gozes Yeheskel Pasmanik-Chor Activity-Dependent Neuroprotective Protein (ADNP) A Case Study for Highly Conserved Chordata-Specific Genes Shaping the Brain and Mutated in Cancer J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
httpwwwadnpgenecom Feb (0) doi 101038ng2899 Nat Genet 46380-384
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Bioinformatics The ADNP Network
Gozes Yeheskel Pasmanik-Chor Activity-Dependent Neuroprotective Protein (ADNP) A Case Study for Highly Conserved Chordata-Specific Genes Shaping the Brain and Mutated in Cancer J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
httpwwwadnpgenecom Feb (0) doi 101038ng2899 Nat Genet 46380-384
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
httpwwwadnpgenecom Feb (0) doi 101038ng2899 Nat Genet 46380-384
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Gozes Yeheskel and Pasmanik-Chor J Alzheimers Dis 2015 Jan 145(1)57-73 doi 103233JAD-142490
Specific to Chordata
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
0010203040506
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP
Males Females
Rela
tive m
RN
A c
onte
nt
00
1
2
3
4
5
Postmortem ADNP
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleelF4E
0
005
01
015
02
025
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleADNP2
0
5
10
15
20
25
ADNP++ ADNP+-
2^-
dC
t
Male
FemaleAPOE
0
05
1
15
2
25
ADNP++ ADNP+-
2^-
dC
t
Male 9 month oldelF4E
Sexual Dichotomy Gene Expression
Human six-month-old mice
Malishkevich A Amram N Hacohen-Kleiman G Magen I Giladi I Gozes I Activity-Dependent Neuroprotective Protein (ADNP) Exhibits Striking Sexual Dichotomy Impacting on Autistic and Alzheimers Pathologies Transl Psychiatry 2015 Feb 35e501 doi 101038tp2014138
Behavior
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Serum Proteomics Only ADNP Decreases in Alzheimer
Fernandez-Montesinos R Torres M Baglietto-Vargas D Gutierrez A Gozes I Vitorica J amp Pozo D (2010) Activity-dependent neuroprotective protein (ADNP) expression in the amyloid precursor proteinpresenilin 1 mouse model of Alzheimers disease J Mol Neurosci 41 114-120 J Proteomics 2012 Jun 2775(12)3617-29 doi 101016jjprot201204017 Activity-dependent neuroprotector homeobox protein A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease Yang MH et al Alzheimerrsquos disease (AD) serum proteins were analyzed using two-dimensional gel electrophoresis combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MSMS) followed by peptide fragmentation patterning Results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Tangle-like pathology (not seen in healthy brain) ADNP decorates microtubules (healthy)
J Pharmacol Exp Ther 2007323438-434 Neuron Glia Biol 2004 Aug1(3)193-199
ADNP Haploinsufficiency Modelling Tauopathy
ADNP partial deficiency a window to the mouse brain
Healthy Sick ADNP-deficiency results in brain ndash cell death (green ndash fluoro-Jade)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Davunetide (NAP) Discovery
NAP = NAPVSIPQ
Our papers J Neurochem 1999 72(3)1283-93 J Mol Neurosci 2000 1461-8 J Biol Chem 2001 276708-14 Curr Alzheimer Res 2009 6455-60 J Mol Neurosci 2008 35127-41 J Neurochem 2008105537-45 J Biol Chem 200728234448-56 Dev Biol 2007303814-24 Brain Res Dev Brain Res 200314483-90 Curr Pharm Des 201117(10)1040-4 Curr Pharm Des 2011 17 (25) 2603-12 Curr Pharm Des 2011 17 (31) 3413-7 PLoS One 20127(12)e51458 Eur J Pharmacol 20096189-14 Int J Alzheimers Dis 20122012
gt Calculated molecular mass for ADNP-1235628 Da
gt A large protein (limited bioavailability)
gt Davunetide NAPVSIPQ is the smallest active fragment of ADNP which provides brain bioavailability and nerve cell protection
O2-Glucose-Deprivation +NAP
Protecting Microtubules
ADNP Contains a small neuroprotective epitope Protecting against apoptosis
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
NAP Complements ADNP (+-) Cognitive Deficiency
Morris water maze test of brain function
J Pharmacol Exp Ther 2007323438-449
0
10
20
30
40
50
60
70
80
90
100
day1 day2 day3 day4 day5
late
ncy (
sec)
ADNP++
ADNP+-
ADNP+- NAP
gt ADNP-deficient mice (+-) display impaired learning and memory in the Morris water maze (latency to reach the hidden platform is not reduced after training)
gt Impairment is significantly reversed by treatment with NAP
gt Parallel protection against tau pathology is seen also in models of Alzheimerrsquos disease and related neurodegenerations (tau mutant mice)
Tau pathology NAP protects
Neurobiol Dis 2009 May34(2)381-8
Tau mutant mice
mutant mice A small protecting snippet against protein deficits
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Microtubules are Essential Within the Nerve Cell
Neurodegeneration gt Destabilization and breakdown of microtubules
gt Tau hyperphosphorylation
gt Progressive loss of function
gt Leads to cell death
Microtubules
Essential for neuronal structure and function
Neuroprotection gt Davunetide (NAP) crosses the human blood brain
barrier
gt Reduces Tau hyperphosphorylation
gt Stabilizes and repairs microtubules
gt Enhances dendritic spine formation
gt Restores neuronal structure and function
Gozes I Littauer UZ Tubulin microheterogeneity increases with rat brain maturation Nature 1978 276 411-13 Gozes I Sweadner KJ Multiple tubulin forms are expressed by a single neurone Nature 1981 294 477-80 Gozes I Barstable CJ Monoclonal antibodies that recognize discrete forms of tubulin Proc Natl Acad Sci USA 792579-83 Gozes et al Curr Alzheimer Res 2009 6455-60 OzhellipGozes Mol Psychiatry 2014 Oct19(10)1115-24
MAP2
Spines connections
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Microtubules End Binding Proteins Providing Microtubule Dynamics
NAPח
NAP increases microtubule end binding protein association with tubulin
NAP increases ADNP Association with microtubule end binding proteins
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
ldquoMolecules that make our mindsrdquo
EB3
Micro
tub
ule
EB3
EB3+1
p140Cap (SIP)
SrcFyn
NM
DA
R
P
EB31
P
TAU
CLASP2 (SxIP)
APC
APC2 (SSIP)
APC2 EB3
Drebrin
ADNP (SIP)
NAP
The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding (EB) Proteins
Oz S Kapitansky O Ivashco-Pachima Y Malishkevich A Giladi E Skalka N Rosin-Arbesfeld R Mittelman L Sege O Hirsch JA and Gozes I The NAP-Motif of Activity-Dependent Neuroprotective Protein (ADNP) Regulates Dendritic Spines through Microtubule End Binding Proteins Mol Psychiatry 2014 Oct19(10)1115-24
+NAP
NAPADNP increases synaptic
connections through EB (green)
Synapse Formation
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Micro
tub
ule
EB3 EB3
EB3+1
NAP
EB31 EB3
ADNP (NAP)
Synaptic plasticity
NAP ADNP (NAP)
LC3
Autophagosome
LC3 LC3
LC3
LC3 LC3
NAP ADNP (NAP)
bbbbbbbbbbb
Bridging Autopahgy and Microtubules
Immunoprecipitation (IP) of ADNP reveals direct
interaction with microtubule associated protein 1
light chain 3 (LC3B) a major regulator of
autophagy by Western analysis The ADNP
LC3B interaction correlates with the level of
ADNP expression a strong interaction in normal
mice (ADNP++ right band) and a weaker
interaction in ADNP+- mice (left band) The
interaction of ADNP with LC3B was
augmented in the presence of NAP
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Esteves AR Gozes I Cardoso SM (2014) The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease Biochim Biophys Acta 1842 7-21
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
NAP Enhances Microtubule Invasion to the Tip of the Nerve Fortifying Cell Communication
Calculation = Area occupied by dynamic (+Tyr)-tubulin (red left grey right) in the distal-peripheral growth cone (ROI)
PLoS One 20127(12)e51458
Invasion of dynamic microtubules
Increases microtubule dynamics at the tip of the growth cone
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Enlisting Tau (Ties) Back to the Microtubules (Train Tracks)
gt NAP increases the tubulin and tau content of microtubules under conditions of zinc intoxication (note zinc accumulation is a prominent feature of Alzheimerrsquos disease pathology)
S Oz Y Ivashco-Pachima and I Gozes PLoS One 20127(12)e51458
NAP enlists tau to microtubules
S Quraishe C M Cowan and A Mudher University of Southampton Southampton Molecular Psychiatry 2013 EPUB
By enlisting taubuilding microtubules NAP provides protection against impairments in material (axonal) transport in nerve cells ndash key to brain communication
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
24 hours 15 min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min 3min
Colchicine
microtubule
disruption
Anesthesia and
MnCl2 or water
administration
NAP (colchicine
group) and
control placebo
Animal transfer
to magnet
3 Control
acquisitions 11 Acquisitions for estimation of axonal
transport (Manganese contrast)
07
08
09
1
11
12
13
14
15
12 15 18 21 24 27 30 33 36 39 42 45 48
Norm
aliz
ed
sig
na
l in
ten
sity
Time (min)
NAP Prevents Axonal Transport Deficits Mouse
Y Jouroukhin R Ostritsky Y Assaf G Pelled E Giladi and I Gozes Neurobiology of Disease 2013 5679-94
Protection against axonal impairment is coupled to protection against tau pathology in an ALS model
State of the art measurements of axonal transport inhaled MnCl2 provides an MRI contrasting agent (white below) Measurements of progression in the olfactory bulb show disruption by microtubule breakdown (colchicine) and protection by NAP
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
-08
-06
-04
-02
0
02
04
06
08
Normal DD Normal NAP Heterozygous
DD
Heterozygous
NAP
D 2
Cognitive Improvement in a Schizophrenia Model
DD=placebo Heterozygous= Schizophrenia model
Working Hypothesis
gt The stable tubule-only polypeptide (STOP) knockout mice have been shown to provide a valid model for schizophrenia
gt The capacity of the mice to discriminate between novel object and familiar object was calculated as discrimination capacity D2
gt The ldquoschizophrenicrdquo mice (microtubule-deficient) show markedly decreased memory
gt NAP ameliorates this deficiency
Microtubule dysfunction in schizophrenia Merenlender-Wagner et al Peptides 2010311368 Gozes Peptides 201132428 Olson Prize Molecular Psychiatry 2013 Dec 24 doi 101038mp2013174 Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Insert
STOP++ STOP+-
BE
CN
1
NAP treatment reverses BECN1 reduction in STOP+- mouse hippocampus
The STOP+- Mouse Model Mimics Schizophrenia NAP Compensation
Beclin1
MerenlenderndashWagner A Shemer Z Touloumi O Lagoudaki R Giladi E Andrieux A Grigoriadis NC and Gozes I (2014) New horizons in schizophrenia treatment Autophagy 2014 Dec 210(12)2324-32 doi 104161155486272014984274
Beclin1
Cleaning and
recycling system
LC3
LC3
LC3
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Significant Improvement in Schizophrenia Patients
-30
-20
-10
0
10
20
30
40
50
placebo 5 mg QD 15 mg BID placebo 5 mg QD 15 mg BID
Daily Activities Change-from-Baseline
T-Sc
ore
(C
FB)
Week 6 Week 12
Conclusion
Statistically significant treatment effect of davunetide (NAP) 5 mg once daily -QD (p=0023) and combined treatment groups (high dose twice daily ndash BID + low dose once daily) versus placebo (p=0015) (mixed model ANCOVA)
No statistical significance between davunetide groups
Associated with in vivo neuroprotection
Javitt et al (2012) Effect of the neuroprotective peptide davunetide (AL-108) on cognition and functional capacity in schizophrenia Schizophr Res 136 25-31
Jarskog et al(2013) Effects of Davunetide on N-acetylaspartate and Choline in Dorsolateral Prefrontal Cortex in Patients with Schizophrenia Neuropsychopharmacology 38 1245-52 Original support NIA NIMH ISOA Allon Therapeutics
Translation into clinical trials
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
ALZHEIMERrsquoS DISEASE
Alois Alzheimer
Auguste D
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
-20
0
20
40
60
80
100
0 4 8 12 16
Week
DM
TS
12 s
ec d
ela
y (
C
FB
)
placebo 5 mg QD 15 mg BID
p=0067
p=0039
p=0064p=0038
Drug administration Follow-up
Davunetide (15 mg x2day) Improves Memory in aMCI Patients
Statistically significant dose dependent and durable impact seen on memory
change from background (CFB) delayed matching to sample (DMTS)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Michael J Fox
Ronald Reagan
John Nash from the film ldquoA Beautiful Mindrdquo
Lou Gehrig Maurice Ravel Margaret Thatcher
Muhammad Ali
We Fight Against Their Disease
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Autism Schizophrenia and Alzheimerrsquos Disease
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Jarskog et al
Neuropsychopharmacology
2013 Jan 16 doi
101038npp201323 [Epub
ahead of print]
Effects of Davunetide on N-
acetylaspartate and Choline in
Dorsolateral Prefrontal Cortex
in Patients with Schizophrenia
Javitt et al
Schizophr Res 2012
Apr136(1-3)25-31 doi
101016jschres201111001
Epub 2011 Dec 12
Effect of the neuroprotective
peptide davunetide (AL-108)
on cognition and functional
capacity in schizophrenia
Clinical Translation
Medical Validation by
Others
A Glimpse at Our Recent Work
The rescue of microtubule-dependent traffic recovers mitochondrial function in Parkinsons disease AR Esteves I Gozes SM Cardoso
Biochim Biophys Acta 2014 Jan1842(1)7-21
MerenlenderndashWagner A Malishkevich A Shemer Z Udawela M Gibbons A Scarr E Dean B Levine J Agam G and Gozes I (2013) Autophagy plays a key role in the pathophysiology of schizophrenia Mol Psychiatry 2015 Feb20(1)126-32 doi 101038mp2013174 Epub 2013 Dec 24
Molecular Psychiatry Translational Psychiatry Autophagy and Pharmacology Research amp Perspectives 2014 2015
Thanks
Tel Aviv University (TAU)
Thanks
Tel Aviv University (TAU)
Tel Aviv University (TAU)