Dr Rousselot - Paris 7

TARGETED THERAPIES IN HAEMATOLOGY, MECHANISMS OF RESISTANCE
AND THERAPEUTIC APPLICATIONS IN CHRONIC MYELOID LEUKEMIAS.
HEMATOLOGY – IMMUNOLOGY – THERAPEUTIC TARGETS AND CIB
(Biomedical Investigattion Center) - UMR-S-940
LIST OF MEMBERS
Team Leader: Dr Philippe ROUSSELOT
Tel: 01 39 63 86 22 or 01 39 63 94 08
Secretariat: Jacqueline Anais
Tel.: 01 53 72 40 18
Fax: 01 53 72 40 16
Stéphane Prost, PhD student, University Paris 7, CEA
Francis Relouzat, CEA
Benjamin Maneglier, UVSQ
RESEARCH FOCUS
The group has a long experience in the management of Chronic Myeloid Leukemia
(the Group Leader, Philippe Rousselot is vice president of the FILMC, French
Intergroup on Chronic Myeloid Leukemia). CML represents a model for targeted
therapy in hematology and oncology. Tyrosine kinase inhibitors, the leader being
imatinib mesylate block the deregulated kinase activity of BCR-ABL and thus control
the disease. These results lead to the unprecedent improvement of patient outcome ;
However, many questions remain unanswered such as the quick characterization
and the management of patients resistant ou non tolerant to imatinib, the place of
second generation treatments such as nilotinib or dasatinib, the persistence in the
majority of patients of detectable minimal residual disease, the possibility to one day
eradicate the disease and subsequently stop treatment.

These questions raise the more general issue of the intrinsic mechanisms of
resistance or acquired by the tumor cell with targeted therapies. The group aims at
studying the resistance along 3 fields
- Genetic aspects of resistance
- Pharmacological aspects of resistance
- Cellular aspects of resistance
These studies will be performed in vitro and in vivo in preclinical models and in novel
therapeutic trials. The group collaborates with Rose Ann Padua’s team for the in vivo
model; The group is member of the cancer stem cell network Cancéropôle Ile de
France.
MAIN CONTRIBUTIONS
- Using these in vitro approaches, the group has shown that modulation of
STAT5 expression in CD34+ CML cells enhances the action of imatinib on the
stem cell compartiment (collaboration avec le CEA, (PhD subject of S. Prost)
- It has lead to a clinical trial (ACTIM) to test the in vivo efficacy of agents
modulating STAT5 expression (glitazones) in association with imatinib in CML
patients with major cytogenetic response. (Grant from ARC)
- First pharmacological academic study of dasatinib in CML –collaboration with
the Department of Pharmacology of Bordeaux (M Molimard).
CONTROLLED TECHNIQUES
Normal and leukemic cell culture. Viral transduction- Imatinib dosage, DNA
sequencing, animal models
PUBLICATIONS
1. Rousselot, P., B. Hardas, A. Patel, F. Guidez, J. Gaken, S. Castaigne, A.
Dejean, H. de The, L. Degos, F. Farzaneh, and . 1994. The PML-RAR alpha
gene product of the t(15;17) translocation inhibits retinoic acid-induced
granulocytic differentiation and mediated transactivation in human myeloid
cells. Oncogene 9:545-551.
2. Rousselot, P., S. Labaume, J. P. Marolleau, J. Larghero, M. H. Noguera, J. C.
Brouet, and J. P. Fermand. 1999. Arsenic trioxide and melarsoprol induce
apoptosis in plasma cell lines and in plasma cells from myeloma patients.
Cancer Res. 59:1041-1048.

3. O'Brien, S. G., F. Guilhot, R. A. Larson, I. Gathmann, M. Baccarani, F.
Cervantes, J. J. Cornelissen, T. Fischer, A. Hochhaus, T. Hughes, K. Lechner,
J. L. Nielsen, P. Rousselot, J. Reiffers, G. Saglio, J. Shepherd, B. Simonsson,
A. Gratwohl, J. M. Goldman, H. Kantarjian, K. Taylor, G. Verhoef, A. E. Bolton,
R. Capdeville, and B. J. Druker. 2003. Imatinib compared with interferon and
low-dose cytarabine for newly diagnosed chronic-phase chronic myeloid
leukemia. N.Engl.J.Med. 348:994-1004.
4. Larghero, J., T. Leguay, S. Mourah, I. Madelaine-Chambrin, A. L. Taksin, E.
Raffoux, J. N. Bastie, L. Degos, P. Berthaud, J. P. Marolleau, F. Calvo, C.
Chomienne, F. X. Mahon, and P. Rousselot. 2003. Relationship between
elevated levels of the alpha 1 acid glycoprotein in chronic myelogenous
leukemia in blast crisis and pharmacological resistance to imatinib (Gleevec)
in vitro and in vivo. Biochem.Pharmacol. 66:1907-1913.
5. Rousselot, P., J. Larghero, B. Arnulf, J. Poupon, B. Royer, A. Tibi, I.
Madelaine-Chambrin, P. Cimerman, S. Chevret, O. Hermine, H. Dombret, B. J.
Claude, and F. J. Paul. 2004. A clinical and pharmacological study of arsenic
trioxide in advanced multiple myeloma patients. Leukemia 18:1518-1521.
6. Larghero, J., N. Gervais, B. Cassinat, J. D. Rain, M. H. Schlageter, R. A.
Padua, C. Chomienne, and P. Rousselot. 2005. Farnesyltransferase inhibitor
tipifarnib (R115777) preferentially inhibits in vitro autonomous erythropoiesis of
polycythemia vera patient cells. Blood 105:3743-3745.
7. Cortes, J., P. Rousselot, D. W. Kim, E. Ritchie, N. Hamerschlak, S. Coutre, A.
Hochhaus, F. Guilhot, G. Saglio, J. Apperley, O. Ottmann, N. Shah, P. Erben,
S. Branford, P. Agarwal, A. Gollerkeri, and M. Baccarani. 2007. Dasatinib
induces complete hematologic and cytogenetic responses in patients with
imatinib-resistant or -intolerant chronic myeloid leukemia in blast crisis. Blood
109:3207-3213.
8. Nasr, R., M. C. Guillemin, O. Ferhi, H. Soilihi, L. Peres, C. Berthier, P.
Rousselot, M. Robledo-Sarmiento, V. Lallemand-Breitenbach, B. Gourmel, D.
Vitoux, P. P. Pandolfi, C. Rochette-Egly, J. Zhu, and H. de The. 2008.
Eradication of acute promyelocytic leukemia-initiating cells through PML-
RARA degradation. Nat.Med. 14:1333-1342.
9. Shah NP, Kim DW, Kantarjian H, Rousselot P, Llacer PE, Enrico A, Vela-
Ojeda J, Silver RT, Khoury HJ, Müller MC, Lambert A, Matloub Y, Hochhaus A.
Potent, transient inhibition of BCR-ABL with dasatinib 100 mg daily achieves
rapid and durable cytogenetic responses and high transformation-free survival
rates in chronic phase chronic myeloid leukemia patients with resistance,
suboptimal response or intolerance to imatinib. Haematologica. 2010
Feb;95(2):232-40. PubMed

10. Roth O, Spreux-Varoquaux O, Bouchet S, Rousselot P, Castaigne S,
Rigaudeau S, Raggueneau V, Therond P, Devillier P, Molimard M, Maneglier
B. Imatinib assay by HPLC with photodiode-array UV detection in plasma from
patients with chronic myeloid leukemia: Comparison with LC-MS/MS. Clin
Chim Acta. 2010 Feb;411(3-4):140-6. Epub 2009 Oct 22. PubMed PMID:
19853594.
PUBLICATIONS IN PRESS
1. Shah NP, Kim DW, Kantarjian H, Rousselot P, Llacer PE, Enrico A, Vela-
Ojeda J,
Silver RT, Khoury HJ, Müller MC, Lambert A, Matloub Y, Hochhaus A. Potent,
transient inhibition of BCR-ABL with dasatinib 100 mg daily achieves rapid and
durable cytogenetic responses and high transformation-free survival rates in
chronic phase chronic myeloid leukemia patients with resistance, suboptimal
response or intolerance to imatinib. Haematologica. 2010 Feb;95(2):232-40.
PubMed
PMID: 20139391; PubMed Central PMCID: PMC2817025.
2. Pautas C, Merabet F, Thomas X, Raffoux E, Gardin C, Corm S, Bourhis JH,
Reman
O, Turlure P, Contentin N, de Revel T, Rousselot P, Preudhomme C,
Bordessoule D,
Fenaux P, Terré C, Michallet M, Dombret H, Chevret S, Castaigne S.
Randomized
study of intensified anthracycline doses for induction and recombinant
interleukin-2 for maintenance in patients with acute myeloid leukemia age 50
to
70 years: results of the ALFA-9801 study. J Clin Oncol. 2010 Feb
10;28(5):808-14.
Epub 2010 Jan 4. PubMed PMID: 20048183.
3. Roth O, Spreux-Varoquaux O, Bouchet S, Rousselot P, Castaigne S,
Rigaudeau S,
Raggueneau V, Therond P, Devillier P, Molimard M, Maneglier B. Imatinib
assay by
HPLC with photodiode-array UV detection in plasma from patients with chronic
myeloid leukemia: Comparison with LC-MS/MS. Clin Chim Acta. 2010
Feb;411(3-4):140-6. Epub 2009 Oct 22. PubMed PMID: 19853594.

4. Maury S, Huguet F, Leguay T, Lacombe F, Maynadié M, Girard S, de
Labarthe A,
Kuhlein E, Raffoux E, Thomas X, Chevallier P, Buzyn A, Delannoy A,
Chalandon Y,
Vernant JP, Rousselot P, Macintyre E, Ifrah N, Dombret H, Béné MC; Group
for
Research on Adult Acute Lymphoblastic Leukemia. Adverse prognostic
significance
of CD20 expression in adults with Philadelphia chromosome-negative B-cell
precursor acute lymphoblastic leukemia. Haematologica. 2010 Feb;95(2):324-
8. Epub
2009 22. PubMed PMID: 19773266; PubMed Central PMCID: PMC2817037.
EN PREPARATION
1. Discontinuation of imatinib in patients with chronic myeloid leukaemia who
have maintained a complete molecular response for at least 2 years: the
multicentre Stop Imatinib (STIM) trial. François-Xavier Mahon, Delphine Réa,
Joëlle Guilhot, François Guilhot, Françoise Huguet, Dr Franck Nicolini,
Laurence Legros, Aude Charbonnier, Agnès Guerci, Bruno Varet, Gabriel
Etienne, Josy Reiffers and Philippe Rousselot. On behalf of the Intergroupe
Français des Leucémies Myéloïdes Chroniques (FILMC). Accepted Lancet
Oncology.
2. Frequency of molecular responses to imatinib plus Pegylated form of
Interferon-alpha2a in newly diagnosed chronic phase chronic myeloid
leukaemia. Preudhomme C, Guilhot J, Nicolini F, Rigal-Huguet F, Rousselot
P,Vekoff A , Legros l, Guerci A, Coiteux V, Rea D, Maloisel F, Gardembas M,
Berthou C, Jourdan E,Berger M, Corm S, Delmer A, Allard C, Roche-
Lestienne C, Eclache V, Mahon FX and Guilhot F.
3. Accepted New England J Medicine.
4. The addition of Daunorubicin to Imatinib Mesylate in combination with
Cytarabine significantly improves the response rate and the survival of
patients with Myeloid Blast Crisis Chronic Myelogenous Leukemia (AFR01
study).
5. Bénédicte Deau, Franck E. Nicolini, Joelle Guilhot, Fathia Merabet, Françoise
Huguet, Bernard Rio, Agnès Guerci, Laurence Legros, Cécile Pautas,
Delphine Réa, Emmanuel Raffoux, Christian Berthou, Denis Guyotat, Pascale
Cony-Makhoul, Martine Gardembas, Sylvie Castaigne, Mauricette Michallet,
Sandrine Hayette, Jean Michel Cayuela, Isabelle Radford Weiss, François-
Xavier Mahon, François Guilhot and Philippe Rousselot for the French
Intergroup For Chronic Myelogenous Leukemia (Fi-LMC).
6. Down regulation of STAT5 expression induced by glitazones results in a
significant decrease of clonogenic capacities of chronic myelogenous
leukaemia CD34+ cells.

7. Stéphane Prost, Francis Relouzat, Marc Spentchian, Gerald Massonet, Jean
Paul Beressi, Christine Chomienne, Philippe Leboulche and Philippe
Rousselot.