Implementation of erythroid lineage analysis by flow cytometry in diagnostic models for myelodysplastic syndromes

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Publishe Ahea of Print on September 22, 2016, as oi:10.3324/haematol.2016.147843. Copyright 2016 Ferrata Storti Founation.

1 Publishe Ahea of Print on September 22, 2016, as oi: /haematol Copyright 2016 Ferrata Storti Founation. Implementation of erythroi lineage analysis by flow cytometry in iagnostic moels for myeloysplastic synromes by Eline M.P. Cremers, Theresia M. Westers, Canan Alhan, Clauia Cali, Heleen A. Visser-Wisselaar, Dana A. Chitu, Vincent H.J. van er Velen, Jeroen G. te Marvele, Saskia K. Klein, Petra Muus, Eo Vellenga, Georgina E. De Greef, Marie-Cecile C.J.C. Legeur, Pierre W. Wijermans, Marian J.P.L. Stevens-Kroef, Pero a Silva-Coelho, Joop H. Jansen, Gert J. Ossenkoppele, an Arjan A. van e Loosrecht Haematologica 2016 [Epub ahea of print] Citation: Cremers EM, Westers TM, Alhan C, Cali C, Visser-Wisselaar HA, Chitu DA, van er Velen VH, te Marvele JG, Klein SK, Muus P, Vellenga E, De Greef GE, Legeur MC, Wijermans PW, Stevens-Kroef MJ, a Silva-Coelho P, Jansen JH, Ossenkoppele GJ, an van e Loosrecht AA. Implementation of erythroi lineage analysis by flow cytometry in iagnostic moels for myeloysplastic synromes. Haematologica. 2016; 101:xxx oi: /haematol Publisher's Disclaimer. E-publishing ahea of print is increasingly important for the rapi issemination of science. Haematologica is, therefore, E-publishing PDF files of an early version of manuscripts that have complete a regular peer review an have been accepte for publication. E-publishing of this PDF file has been approve by the authors. After having E-publishe Ahea of Print, manuscripts will then unergo technical an English eiting, typesetting, proof correction an be presente for the authors' final approval; the final version of the manuscript will then appear in print on a regular issue of the journal. All legal isclaimers that apply to the journal also pertain to this prouction process.

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29 Supplementary Table S1 The applie 4-color (panel A) an 8-color (panel B) markers. Per antiboy CD number, (clone), an manufacturer are epicte. DAKO: DakoCytomation, Glostrup, Denmark; BD: BD Biosciences, San Jose, CA, USA. BL: BioLegen, San Diego, Ca, USA. Sanquin, Amsteram, The Netherlans. BC: Beckman Coulter, Miami, FL, USA. IS: Immunostep, Salamanca, Spain. Table S1A FITC PE PerCP APC 1 (2D1) BD 2 CD16 (DJ130c) DAKO CD13 (L138) BD CD11b (D12) BD 3 CD34 (8G12) BD CD11b (D12) BD HLA-DR (L243) BD 4 CD36 (CLB-IVC7) Sanquin CD33 (P67.6) BD CD14 (MoP9) BD 5 CD36 CD64 CD14 (10.1) DAKO 6 CD15 (MMA) BD CD10 (SS2/36) DAKO CD34 (8G12) BD 7 CD34 CD117 (104D2) BD CD13 (WM15) BD CD33 (P67.6) BD 8 CD34 9 CD5 CD19 CD34 (DK23) DAKO (SJ25C1) BD 10 CD2 CD56 CD34 (MT910) DAKO (My31) BD 11 CD13 CD7 CD34 (WM-47) DAKO (M-T701) BD 12 CD13 CD25 CD34 (ACT-1) DAKO 13 CD71 (Ber-T9) BD CD235a (JC159) DAKO CD117 (104D2) Dako

30 Table S1B FITC PE PerCP- Cy5.5 PC7 APC APC-H7 V450 KO 1 CD34 (8G12) BD 2 CD16 (DJ130c) DAKO 3 CD2 (MT910) DAKO 4 CD36 (CLB- IVC7) Sanquin 5 CD5 (DK23) DAKO CD13 (L138) BD CD64 (10.1) DAKO CD105 (43A3) BL CD56 (My31) BD CD117 (104D2D1) BC CD34 CD117 CD11b (D12) BD CD34 CD117 IREM2 (UP-H2) IS CD34 CD117 CD33 (P67.6) BD CD34 CD117 CD7 (M- T701) BD CD10 (HI10A) BD CD14 (MoP9) BD CD71 (M- A712) BD CD19 (SJ25C1) BD HLA- DR (L243) BD HLA- DR HLA- DR HLA- DR HLA- DR (J.33) BC 6 CD15 (MMA) BD 7 CD7 (M-T701) BD CD25 (ACT-1) DAKO CD235a (JC159) DAKO CD34 CD117 CD123 (9F5) BD CD34 CD117 CD13 (WM15) BD CD38 (HB7) BD CD71 (M- A712) BD HLA- DR HLA- DR

31 Table 2S Upgrae patients per WHO-Classification after aition of the erythroi evaluation. This table provies the absolute patient counts belonging to Figure 2. WHO-classification Upgrae A > B No MDS Upgrae B > C MDS C MDS after aition erythroi evaluation (group total) RA (4) RARS (20) RCMD (23) RCMD-RS (27) RAEB (14) Del(5q) (12) MDS-U (2) CMML (4)

32 Figure S1 Example of a normal bone marrow (A) an a MDS patient (B) Figure A provies an example of a healthy control. Figure B provies an example of a MDS patient (RCMD-RS) which has an increase percentage of erythroi progenitors (EP). Furthermore, CD71 expression is longer retaine uring maturation. This is reflecte in the increase CV of CD71 an CD36. Finally, the MDS patient shows a ecrease MFI of CD71. This patient scores 4 out of 4 points, revealing clear MDS specific yserythropoiesis as assesse by FC. The otte lines plotte in figure B are the normal reference subtracte from figure A.

33 A EP CV CV EP B CV CV

Phase I/II Study of the Combination of Panobinostat and Carfilzomib in Patients with Relapsed/Refractory Multiple Myeloma

$Phase I/II Study of the Combination of Panobinostat and Carfilzomib in Patients with Relapsed/Refractory Multiple Myeloma$ Publishe Ahea of Print on February 20, 2015, as oi:10.3324/haematol.2014.119735. Copyright 2015 Ferrata Storti Founation. Phase I/II Stuy of the Combination of Panobinostat an Carfilzomib in Patients with