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FAP ‐ Genetics
Familial autosomic dominant.
Familial autosomic dominant.
No modifier genes identified so far, but familial
No modifier genes identified so far, but familial aggregation of subphenotypes.
aggregation of subphenotypes.
Parent of origin effects: Anticipation of age at onset, increased penetrance, particularly on mother to child transmission. Still unexplained
enetic im rintin ? .
Parent of origin effects: Anticipation of age at onset, increased penetrance, particularly on mother to child transmission. Still unexplained
enetic im rintin ? .
?I
II
III
IV
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II – Molecular Biology
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–
The fundamental constituent of amyloid in PAF is identical
immunochemically to a small serum
The fundamental constituent of amyloid in PAF is identical
immunochemically to a small serumprotein, transthyretin (TTR).
[Costa et al., PNAS, 1978]
protein, transthyretin (TTR).
[Costa et al., PNAS, 1978]
oc em ca c arac er za on o sfundamental constituent of amyloid
reveals the presence of a TTR mutation,
oc em ca c arac er za on o sfundamental constituent of amyloid
reveals the presence of a TTR mutation, w a me on ne or va nesubstitution at position 30.w a me on ne or va nesubstitution at position 30.
[Saraiva et al., Trans Assoc Am Physicians, 1983][Saraiva et al., Trans Assoc Am Physicians, 1983]
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-20 -10
ACAGAAGTCCACTCATTCTTGGCAGGATGGCTTCTCATCGTCTGCTCCTCCTCTGCCTTGCTGGA
FAP..........................-M--A--S--H--R--L--L--L--L--C--L--A--G-
-1 1 10
CTGGTATTTGTGTCTGAGGCTGGCCCTACGGGCACCGGTGAATCCAAGTGTCCTCTGATGGTCAAA
-L--V--F--V--S--E--A--G--P--T--G--T--G--E--S--K--C--P--L--M--V--K-
20 30
ATTR V30MGTTCTAGATGCTGTCCGAGGCAGTCCTGCCATCAATGTGGCCATGCATGTGTTCAGAAAGGCTGCT
-V--L--D--A--V--R--G--S--P--A--I--N--V--A--M--H--V--F--R--K--A--A-
40 50
GATGACACCTGGGAGCCATTTGCCTCTGGGAAAACCAGTGAGTCTGGAGAGCTGCATGGGCTCACA-D--D--T--W--E--P--F--A--S--G--K--T--S--E--S--G--E--L--H--G--L--T-
60 70 80
ACTGAGGAGGAATTTGTAGAAGGGATATACAAAGTGGAAATAGACACCAAATCTTACTGGAAGGCA
-T--E--E--E--F--V--E--G--I--Y--K--V--E--I--D--T--K--S--Y--W--K--A-
90 100
CTTGGCATCTCCCCATTCCATGAGCATGCAGAGGTGGTATTCACAGCCAACGACTCCGGCCCCCGC
-L--G--I--S--P--F--H--E--H--A--E--V--V--F--T--A--N--D--S--G--P--R-
110 120CGCTACACCATTGCCGCCCTGCTGAGCCCCTACTCCTATTCCACCACGGCTGTCGTCACCAATCCC
-R--Y--T--I--A--A--L--L--S--P--Y--S--Y--S--T--T--A--V--V--T--N--P-
127
AAGGAATGAGGGACTTCTCCTCCAGTGGACCTGAAGGACGAGGGATGGGATTTCATGTAACCAAGA
-K--E--*-.........................................................
GTATTCCATTTTTACTAAAGCAGTGTTTTCACCTCATATGCTATGTTAGAAGTCCAGGCAGAGACA
..................................................................
ATAAAACATTCCTGTGAAAGGCACTTTTCATTCC
...................................
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Amyloid Type Precursor Disease
apo econ ary reac ve amy o os sFMF
AL Ig light chains (κ,λ)
ATTR Transthyretin
TTR V30M
TTR L111M
Senile systemic amyloidosis
Polyneuropathy
Cardiomiopathy
AapoAI apoA‐I Arg 26
apoA‐I Arg 60
Polyneuropathy, Nephropathy
Nephropathy
A Lys Lysozime Thr 56, His 67 Nephropathy
AFibA Fibrinogen Aα E526V, R554L Nephropathy
AGel Gelsolin Asn 187 FFA
ACys Cystatin C Gln 68 HCHWA, Icelandic type
Aβ2M β2‐microglobulin Hemodialisis amyloidosis
’
βPP Gln 618
HCHWA, Dutch type
AScr PrPC, PrPSc, PrPCJD Kuru, Scrapie, CJD, GSS
m n a etes me tus, type
ACal procalcitonin Thyroid medulary carcinoma
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u a ons
Arg 47 Ile 50
Glu 18
Neuropathy
Vitreous/other
Lys 61
Val 107Leu 30
Leu 33
Val 33
Tyr 77
Gly 42
Ala 30 Ile 33
Arg 58
Cys 114
ro
Gly 18
Asn 35Ser 24
Thr 34
ro
Ala 47Val 47
Gln 89 Ala 60
Arg 10
Ala 49
Pro 55
Gly 30Met 30
Arg 50
Leu 64
Gly 97
Ile 112
Ile 122
Leu 68
Lys 59
Asp 45
Ser 84
Ala 71
His 60
Ile 20 Met 111Ile 50
Ser 125 Ser 6
Cardiomiopathy
Met 119
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III – Epidemiology
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PAF ‐ distribuição
‐Cávado
(733)
e porta ores nut3
100 ou mais (4)
60 a 99 (3)
40 a 59 (3)
30 a 39 (3)
3525 TTR V30M carriers registeredat Centro de Estudos de
Paramiloidose, Porto (June 2008),
3525 TTR V30M carriers registeredat Centro de Estudos de
Paramiloidose, Porto (June 2008), 78
Serra da Estrela
(1206)Gran e Porto
(102)
Baixo Vouga
10 a 19 (2)
0 a 9 (11)
Junho de 2008
covering most patients an eat ycarriers above 20 years of age (data from genetic counceling).
covering most patients an eat ycarriers above 20 years of age (data from genetic counceling).
(261)Baixo Mondego
(72)Cova da Beira
The residence of 97% of theseThe residence of 97% of thesecarriers is known. Northern coastalareas most heavily affected.carriers is known. Northern coastalareas most heavily affected. Península de Setubal
(90)
(291)ran e s oa
Number of live patients > 1000. National re istr im lementationNumber of live patients > 1000. National re istr im lementationunder way.under way.
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‐
Northern Portugal, one study only (HIEF)
Alves IL et al. Human Mutation 1997 9 3 226‐33
Northern Portugal, one study only (HIEF)
Alves IL et al. Human Mutation 1997 9 3 226‐33
In 5000 individuals:In 5000 individuals:
• Met 30 8
• Met 119 35
• Met 30/Met 119 1
• Asn 90 12
• Unknown 3
• Thr 190 1
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‐
Northern Sweden – endemic counties (ELISA)
Holmgren et al. J Med Genet 1994, 31, 351‐4
Northern Sweden – endemic counties (ELISA)
Holmgren et al. J Med Genet 1994, 31, 351‐4
In 1276 individuals:In 1276 individuals:
• Met 30 16
• Met 30/Met 30 3
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–
© 2001 National Geographic Society
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The Travels of a Gene?
© 2001 National Geographic Society
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-
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• Nerve and skin
• Fat as irate
• Other
The basis of diagnosis in old times (before genetic testing)The basis of diagnosis in old times (before genetic testing)
It is rarely positive in asymptomatic carriersIt is rarely positive in asymptomatic carriers
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ATTR V30M – Molecular Testin
30
·· AGTCCTGCCATCAATGTGGCCGTGCATGTG ··
·· -S--P--A--I--N--V--A-- V --H--V- ··
↓ ·· AGTCCTGCCATCAATGTGGCC A TGCATGTG ··
·· -S--P--A--I--N--V--A-- M --H--V- ··
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–
AlternativesAlternatives
• SSCP
• DHPLC
TTR V28M• e c.
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-
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FAP - Tr tm nt
Before the advent of liver transplantation, FAP was anincurable disease.Before the advent of liver transplantation, FAP was anincurable disease.
,
and is effective because > 90% of human TTR is producedin the liver.
,
and is effective because > 90% of human TTR is producedin the liver.
Only current alternative:Only current alternative:, ,
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–
Aconselhamento genético
• Diagnóstico pré‐natal
• Dia nóstico ré‐im lanta ão
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DESENVOLVIMENTO
EMBRIONÁRIO PRÉ- IMPLANTAÇÃO
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1º Dia Dia 1-2 Dia 2
Dia 3
• 1º e 2º glóbulos polares
Dia 4
BIÓPSIA
• a ast meros, ao ia
• Células da trofoectoderme de blastocistos, ao 5º dia
Dia 4
Dia 5
Dia 6ia 6-7
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PROCESSO DE FIV
• Exames preliminares ♀ ♂
≥ • ≥
• ICSI - não contaminaçãoez
c u as a granu osa
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ABERTURA DA ZONA PELÚCIDA
• Solução ácida de Tyrode (pH 2.2)• aser• Dissecção parcial
*Meio sem Ca2+/Mg2+
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TRANSFERÊNCIA DE EMBRIÕES
4º ou 5º dia após punção folicular
TAXA DE GRAVIDEZ CLÍNICA EM DGPI22,4%
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• Inibição da formação de amilóide
• Estabilizadores do tetrâmero de TTR • Remoção da proteína circulante
• Remoção dos depósitos
• Antagonistas dos glicosaminoglicanos
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r og nese ‐.
Estes estudos mostraramue é relativamente fácil
obter fibras com ascaracterísticas genéricas dasfibras de amilóide a partir deum grande número deproteínas ou fragmentos
proteicos.
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r og nese ‐A fibrilogénese passa quase sempre pela formaçãode um intermediário, que pode formar fibras deimediato, ou a partir de agregados de tipo amorfo.
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-pH 5.3 pH <3pH 3
Rearranjedtetramer
Nativetetramer A-state
Amyloid
fibrils
Early model, based onacidic denaturationEarly model, based onacidic denaturation
SAP GAGs
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Serag et al. Nature Struct Biol 9: 734-9, 2002
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‐
Hou, X. et al. FEBS Journal 2007, 274 (7), 1637-50.
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Kadowaki et al., J Chem Neuroanatomy 28 (2004) 93–100
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FAP – lini l Tri l
DMSO CEP II/III (?) 1984
Apheresis CEP I/II 2001
CPHPC (Ro 63‐8695) CAAPP / Roche II/III 2002
Fx‐1006A FoldRx II/III 2009
un sa
Doxycyclin IBMC ? ?