3 rd International ELN Workshop: Standardization of Flow Cytometry in MDS London, Greenwich 4-6 November 2010

a.a. van de loosdrecht, chair r. ireland, co-chair update ELNet wp8; ASH Orlando, December 4, 2010 and ELNet WP conference, Mannheim, February 1, 2011

VU University Medical Center Amsterdam The Netherlands

3 rd International ELN Workshop: Standardization of Flow Cytometry in MDS London, Greenwich 4-6 November 2010

a.a. van de loosdrecht, chair r. ireland, co-chair

12 countries; 28 participants; in- and outside Europe • • • • • • • • • • • • • Austria: P. Valent France: MC Bene, J. Feuillard Germany: W. Kern Great Britain: R. Ireland, U. Johansson, M. Cullen, G. Mufti, T. Minle, S. Kordasti Italy: M. della Porta [absent] Netherlands: C. Alhan, A.A. van de Loosdrecht, J te Marvelde, V. van de Velden, TM Westers, B. Moshaver, T deWitte Spain: [absent] Sweden: A. Porwit Greece: K. Psarra

Japan:

K. Ogata

USA:

D. Wells, M. Loken, J Cutler

Australia:

K. Burbury VU University Medical Center Amsterdam The Netherlands

Goal:

• • define minimal flow cytometric criteria to analyse MDS  ELN 2011 [2 consensus documents: rationale/technical] • Define: role of FCM: – Diagnosis – Prognosis – predictive value for R/ [emerging new drugs] – disease monitoring VU University Medical Center Amsterdam The Netherlands

Proposed antigens of importance in FCM in MDS:

ELN consensus 2009

ELN consensus document MDS and flow cytometry: van de Loosdrecht, ELN members WP8, et al., Haematologica 2009;94:1124-1134

VU University Medical Center Amsterdam The Netherlands

BONE MARROW SUB SET RECOMMENDED ANALYSIS Overview of some recommended flow cytometric analyses of

Relation CD71 and CD235a

subsets in the BM by the ELNet; London 2010

Expression of CD71 Immature myeloid and monocytic progenitors Granulocytes Monocytes Progenitor B cells % of cells CD34 + CD45 + in combination with FCS/SSC; expression of CD13 and CD117; expression of HLA-DR expression of CD5, CD7, CD56; asynchronous expression of CD11b, CD15 % of cells SSC as ratio vs. SSC of lymphocytes relation of CD13 and CD11b relation of CD13 and CD16 expression of CD10 on mature neutrophils ?? % of cells relation of HLA-DR and CD11b relation of CD36 and CD14 expression of CD13 and CD33; expression of CD56 enumeration as fraction of total CD34+ VU University Medical Center Amsterdam The Netherlands

Analysis of a minimized multiparameter panel

% CD34 + myeloblasts % CD34 + B cells of total CD34 + CD45 expression of Ly vs. Myeloblasts SSC ratio of CD10 neutrophils vs. Ly

Ogata et al, Blood, 2006; Ogata/Della Porta, Haematologica, 2009 Della Porta, et al., 2011 (to be submitted); Japan/Italy/Netherlands/Sweden/USA/UK

VU University Medical Center Amsterdam The Netherlands

Analysis of myelomonopoiesis

severe April 1, 2010 mild normal

n=45 n=22 n=58 n=24 n=45 n=34 n=31 n=37

Wells et al., Blood, 2003; Van de Loosdrecht and Westers et al., Blood, 2008 Alhan et al.; unpublished data set june 2010 Chu et al., personal communication ASH2010; Kern et al., Cancer 2010

VU University Medical Center Amsterdam The Netherlands

FCSS is related to transfusion-dependency and progression to AML

low/int-I risk MDS

myeloblasts either: CD5 ( o ), CD7 ( o ) or CD56 ( o ) positive In part published: Loosdrecht AA van de, Westers TM et al., Blood 2008;111 Chu et al., personal communication; ASH#2010

VU University Medical Center Amsterdam The Netherlands

Clinical impact of flow cytometry in MDS

1. Flow cytometry is recognized in the diagnostic and prognostic work-up of patients with cytopenia (ELN-2009; ELN-2011) 2. An ELN consensus guideline for flow cytometry in MDS is now available (ELN, Haematologica, 2009; updated 2011; to be submitted) 3. Rationale for clinical use of flow cytometry is defined (ELN-2011: to be submitted) VU University Medical Center Amsterdam The Netherlands

Perspectives: 2011

 • Submission 1q/2011: 3 ELN consensus documents • 4 th international ELN meeting in Pavia

Italy

: co-chair Matteo dellaPorta (October 2011) • • • • •  diagnostics  FCSS-revised  erythroid-lineage  incorporate software tools for analysis/statistics  clinical reporting VU University Medical Center Amsterdam The Netherlands