Wolf-Karsten Hofmann, MD, PhD Professor of Medicine Department of Hematology and Oncology

1. European LeukemiaNet Work-Package 8: Myelodysplastic Syndromes Suggestions on Microarray Experiments Wolf-Karsten Hofmann, MD, PhD Professor of Medicine Department of Hematology and Oncology University Hospital Benjamin Franklin Berlin, Germany

2. Microarrays in MDS: What is the need? • Multiple microarray-data from CD34+ cells (other cells are in discussion, granulocytes?) at the time of initial diagnosis of MDS. • To correlate specific gene expression profiles with diagnosis and risk/course of the disease? • To add new prognostic parameters/gene expression profiles? • To predict response to treatment? • Central sample collection! • Standardization of microarray protocols (e. g. Affymetrix platform)! • Tight collaboration with WP13 (Gene Profiling: T. Haferlach, W. Hiddemann)!

3. Microarray Experiments in MDS • Aims • To use gene expression profiles for classification or risk-evaluation in MDS • To detect altered molecular pathways in MDS-cells • Technique • Most common: Oligonucleotide microarrays (Affymetrix) • Less common: cDNA-arrays

4. Time course of gene expression (normal CD34+) Erythropoiesis (CD71+) All genes (22,283) Megakaryopoiesis (CD61+) All genes (22,283) WKH 02/2003

5. MicroFluidic Cards Application in MDS studies?

9. (Future) applications Hematology UMCN • Downstream targets of WT1 in AML, MDS, and in murine transplant model • Hematology Card, targets involved in leukemogenesis, cell cyclus, differentiation/proliferation • ABC transporter genes in AML and MDS • Software package for cluster analysis

10. Meeting of the European LeukemiaNet, WP8 (MDS): Salamanca, Nov. 2004Interaction with AML workpackage Listing: European, national studies recruiting both MDS and AML (according to FAB) patients total number of studies: >25 standard induction treatment: >80 % non-intensive treatment: < 20 % studies including patients aged >60 years: ca. 10

11. Meeting of the European LeukemiaNet, WP8 (MDS): Salamanca, Nov. 2004Interaction with AML workpackage Biological agents active in „overlap studies“ of MDS and AML • farnesyl transferase inhibitors • histone deacetylase inhibitors • anti-angiogenic agents • DNA methyltransferase inhibitors • low-dose melphalan

12. Meeting of the European LeukemiaNet, EHA, Geneva June 10, 2004WP8: MDSInteraction with AML workpackage MDS and AML as a biological continuum • blast cut-off lowered over time (50==>30==>20%) • cytogenetics similar in high-risk MDS and sAML, tAML • inclusion of high-risk MDS and bona fide AML patients in same studies

13. % BM blasts 3 3 4 6 40 30 2 5 20 10 5 1 months 3 6 9 12 1) indolent, „low-risk“ MDS 2) evolving, „high-risk“ MDS 3) acute myeloid leukemia (with or without multilineage dysplasia) 4) AML from MDS (> 6 months duration) 5) „RAEB-t“ (FAB classification) = AML (WHO classification) 6) smouldering AML

14. From: B. Deschler et al., „Treatment decision-making in the biological continuum of older patients with myelodysplastic syndrome and acute myeloid leukemia“ (in prep.)

15. Inclusion for DAC studies % BM blasts % BM blasts Cytogenetics 100 90 80 70 60 50 40 30 20 10 AML DAC 00331 Study all except t(15;17) MDS DAC 06011 EORTC Study all poor risk

16. Implementation and coordination of ongoing and new clinical trialsspecific Phase I/II trials (thalidomide, farnesyltransferase inhibitors, arsenic trioxide and others) Fenaux Trials with Hematopoietic growth factors/iron chelators Hellström Anticytokine and signal transduction modulating studies Fenaux DNA hypomethylating studies Lübbert Eradication of the myelodysplastic clone by novel intensive cytotoxic therapy including autologous SCT Sanz/Mufti New forms of allogeneic stem cell transplantation Martino/De Witte Immunosuppression Ganser