Post on 14-Feb-2018
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HEIM
Hematology and Immunology
6-Dec-11
HEIM
Faculty(building D level 0)
Bieghs LiesbethDe Bruyne ElkeDe Smedt AndreaDehui Xu De Zanger RonaldJinsong HuJoost de ter Beerst Marie Lemaire MiguelLub Susanne Maes Ken Menu ElineNur Haneen Seynaeve CarineVan Valckenborgh ElsVanderkerken KarinWillems Angelo
Clinical Hematology
Caron CharlotteDe Becker AnnFostier KarelSchots RikTrullemans Fabienne
Stem cell laboratory(BTC building level 1)
Arras NicoleDe Becker AnnDe Veirman KimXu SongVan Riet Ivan
Laboratory of Hematology
Bakkus MarleenDe Smet DieterDe Waele MarcDemanet ChristianJochmans KristinOrlando ChristelleVan der Meeren SamVanderstraeten JonaVercammen Martine Verheyden SonjaVerloes An
6-Dec-11
Overview of research lines
• Multiple myeloma: basic, translational, clinic
• Thrombophilia: basic, translational, clinic
• Clonal hemopoietic stem cell disorders: translational
• Natural killer cell receptors: translational
• Molecular targets for risk-group stratification in childhood acute lymphoblastic leukaemia: translational and clinic
6-Dec-11
MULTIPLE MYELOMA
� Plasma cell cancer
� Accumulation in the bone marrow
� Paraprotein
� Angiogenesis
� Osteolytic bone disease
� 500 new patients/year
� Older patients
6-Dec-11
INTERACTIONS BM-MM CELLS
CAM-DR
Lemaire et al. Advances in Cancer Research,
2011, 110:19-42.
angiogenesis
Drug resistance
Bone disease
Survival & proliferation of MM cells
Immune system
Within clinical trial context
Panobinostat
Carfilzomib
Pomalidomide
Elotuzumab
Routinely used
Melphalan
Prednisone
Dexamethasone
Adriamycine
Cyclophosphamide
Velcade
Revlimid
Thalidomide
Bendamustine
Caelyx (liposomal adriamycine)
MEDICATION USED FOR THE TREATMENT OF MYELOMA
6-12-2011
6
ChemotherapyCorticosteroids
Proteasome inhibitor
Immunomodulators
HDAC inhibitor
CS-1 antibody
6-Dec-11
*Methylation
*Post-T histon Modifications*mi RNA (in coll. with
T.VandenDriessche)*In vitro and in vivo* Epigenetic regulating agents(Dr. Elke De Bruyne)
*IGF-1*Notch*Hypoxia*MesenchymalStem cells(Prof. I. Van Riet)
*Different sub
Populations(Dr. Els Van Valckenborgh)*CAM-DR-integrines(Dr. Eline Menu)*Nanobodies(Miguel Lemaire-in coll. with T. Lahoutte)
*MDSc(Dr. Els Van Valckenborghin coll. with
J. Van Ginderachter)*iNKT cells(Dr. Eline Menu)*Immune therapy(Dr. R. Schots in
collaboration with K.
Thielemans)
*Participation toPhase II-III studies(Prof. R. Schots)
Myeloma Center Brussels www.myeloma.be
6-Dec-11
Division of Clinical Hematology
Clinical HematologyR SchotsF TrullemansA De BeckerC CaronK Fostier
Clinical Research
= participation in phase II-III clinical trialsMyeloma >> lymphoma/leukemia/transplantation
Translational researchDendritic cell therapy in myeloma (Collaboration with LMCT)Targeting residual cancer cells in myelomaTargeting BM microenvironment in myeloma
Clinical activity
Hospital Day Clinic Stem cell laboratoryI Van Riet
Hematopoietic stem cell transplantation
Research activity
6-Dec-11
Material
Human MM cell lines
Primary patient material
Syngeneic murine model: 5TMM
MATERIAL
�Similar characteristics as human disease
5T2moderate growth (12w)
osteolytic lesions
5T33
rapid growth (3w)
no osteolytic lesions
In vitro cell lines
6-Dec-11
Techniques-in vitro
• Cell culture and isolation (MACS, FACS)
• Proliferation, apoptosis, migration, invasion, clonogenic assays
• Cultures in hypoxic conditions (in collaboration with TROP)
• Flow cytometry: analysis + sorting (in collaboration with LMCT)
• Elisa
• Western blot analysis
• RT-PCR, Real Time PCR, Chip analysis
6-Dec-11
5T2 or 33MM model
0 weeks
Inject 5TMM cells SacrificeMyeloma Disease
Paraprotein
FACS
Angiogenesis
MVD
Bone disease
Techniques: in vivo
- 5T2 or 5T33MM model
- prophylactic or therapeutic treatment
6-Dec-11
BMEC
MM
BMSC
Osteoclast
BO
NE
CCR2
MCP-1
CD
44
v1
O
67kD LR
La
min
in-1
IGF-1R
IGF-I
CD44v6
67kD LR
Osteoblast
DKK1 RANK
RANKL
RANKL
MIP-1αααα
IL-1ββββ
MIP-1αααα
CCR5
VEGFPDGFbFGFAng-1
HGF
CXCR4
SDF-1ααααMMP9adipocyt
lept
in
6-Dec-11
Funding: FWO-Vlaanderen, OZR-VUB (GOA, HOA and projects), Stichting tegen Kanker, Vlaamse Kankerliga, MMRF, IMF, EUFP6, EUFP7
International Collaborations: •P. Croucher (Sheffield, UK)•H. Jernberg-Wiklund (Uppsala, USA)•S. Lentsch (Pittsburgh, USA)•W. Matsui (Baltimore, USA)•D. Hose (Heidelberg, Germany)•B. Klein (Montpellier, France)•S. Sahota (Southampton, UK)•T. Anderson (Danmark)•H. Johnson (Aarhus, Danmark)•X. Leleu (Lille, France)•I. Slaper (Utrecht, The Netherlands)
6-Dec-11
ThrombophiliaThrombophilia
• Acquired or inherited tendency for the development of
venous thromboembolism
• Our interest: inherited thrombophilia
– Clinical presentation: thrombosis at young age, recurrent events,
familial history, concomitant risk factors
– Treatment, prognosis and counselling in function of classification
6-Dec-11
ThrombophiliaCongenital AT and PC deficiency
• Objectives
– Full laboratory assessment of antithrombin (AT) and protein C
(PC) deficiency
– Molecular characterization of mutations in the AT and PC gene
– Investigation of correlation genotype/phenotype
– National and international collaborations as expert centre
for hereditary AT deficiency
• Researchers
– Kristin Jochmans, MD, PhD
– Christelle Orlando, Msc
6-Dec-11
Thrombophilia in assisted reproduction
• Objectives
– Prospective study: influence of thrombophilia on the outcome of
IVF/ICSI in couples with recurrent failure of assisted
reproduction
(implantation failure and miscarriage)
– Protein Z and Protein Z-dependent protease inhibitor as new
parameters in thrombophilia
• Researchers– Kristin Jochmans, MD, PhD
– Christelle Orlando, Msc
6-Dec-11
Thrombophilia Techniques
• Chromogenic substrate assays
• Clotting-based assays
• ELISA
• Immuno-electrophoresis
• Rocket electrophoresis
• PCR
• Sequencing
• MLPA (multiplex ligation-dependent probe amplification)
6-Dec-11
Clonal hemopoietic stem cell
disorders
• Myelodysplastic syndromes and chronic myeloproliferative neoplasms
• Team: M. De Waele, F. Trullemans, K. Jochmans, D. De Smet, S. Van der Meeren,
M. Bakkus
• Analysis of bone marrow aspirates from patients seen in the Department of Clinical Hematology (R. Schots) in the period 2002 -2009
• Immunophenotyping CD34+ cells:
– Question: Diagnostic potential?
– Myelodysplastic syndromes: D. De Smet
– Chronic myeloproliferative neoplasms: S. Van der Meeren
6-Dec-11
Clonal hemopoietic stem cell
disorders
• miRNA profiling:
– Myelodysplastic syndromes:
• miRNA profiling done in UZ Gent (Department of Medical Genetics, F. Speleman, P. Mestdagh)
• Validation and analysis of the data now performed
• Question: Is there a diagnostic miRNA signature for MDS compared to normal bone marrow and secondary cytopenia?
– Chronic myeloproliferative neoplasms:
• miRNA profiling still has to be performed.
• Funding: FWO-Vlaanderen
6-Dec-11
Natural killer cell receptors
• Natural killer cell receptor polymorphism in leukemia and hematopoietic
stem cell transplantation.
• Team: C. Demanet, S. Verheyden
• Analysis of samples of patients seen in the Department of Clinical
Hematology (R. Schots)
• Results:
– The identification of NK cell receptor phenotypes associated with
leukemia
– A defined donor activating NK cell receptor genotype protects against
leukemic relapse after related HLA-identical hematopoietic stem cell
transplantation
– The susceptibility to myeloid and lymphoid leukemia is mediated by
distinct inhibitory KIR-HLA ligand interactions
– The role of the inhibitory KIR ligand HLA-Bw4 and HLA-C expression
levels in the recognition of leukemic cells by natural killer cells.
• Funding: FWO-Vlaanderen
6-Dec-11
Molecular targets for therapy
stratification in ALL
• MRD analyses by specific Ig/TCR real-time PCR on predefined timepoints during therapy
• Prognostic impact of new molecular targets (IKZF, CRFL2)
• Team: – Marleen Bakkus, PhD
– Jona Van der Straeten, Msc
– Emmanuella Kabongo Kanjinga, Msc
– in collaboration with the BSPHO and the EORTC-CLG
• Analysis of bone marrow samples of all Belgian pediatric ALL patients
• Techniques: – Ig/TCR target identification by genescan analysis
– Sequencing of clonal Ig/TCR gene rearrangements
– patient specific design of TaqMan PCR assays
– MLPA (multiplex ligation-dependent probe amplification)
6-Dec-11
Molecular targets for therapy
stratification in ALL
• Funding: WFWG, ‘Stichting tegen Kanker’, ‘Nationaal Kankerplan’
(years)0 1 2 3 4 5
0
10
20
30
40
50
60
70
80
90
100
O N Number of patients at risk :23120 117 112 105 1009 12 7 5 4 4
Overall Logrank test: p<0.0001
EFS %
MRD > 10-2MRD < 10-2
TP1: MRD < 10-2
TP1: MRD > 10-2
VHR
SR
SR if MRD < 10-3 on TP2
VHR if MRD > 10-3 on TP2
No SCT if MRD < 10-3 on TP2
SCT if MRD > 10-3 on TP2
New EORTC-CLG guidelines
Results
EORTC 58881
EORTC 58951
Thank you !!
6-Dec-11