Composition of Cellular Subsets by Flow Cytometry Identifies Differences Between MDS Subtypes and Aplastic Anemia but No Differences Are Identified Between Cases with and without Monosomy 7.
Authors: Mejstrikova Ester , Pelkova Vendula , Reiterová Michaela , Sukova Martina , Zemanova Zuzana , Vodickova Elena , Campr Vit , Michalova Kyra , Jarosova Marie , Formankova Renata , Smisek Petr , Sedlacek Petr , Pospisilova Dagmar , Mihal Vladimir , Hak Jiri , Jabali Yahia , Sterba Jaroslav , Blatny Jan , Zapletal Ondrej , Cerna Zdenka , Votava Tomas , Prochazkova Daniela , Trka Jan , Stary Jan , Hrusak Ondrej 
Published: October 2019

Abstract
Abstract 3802
Poster Board III-738

Introduction
Monosomy 7 or del(7q) are frequent cytogenetic abnormalities in children with myelodysplastic syndrome (MDS) and associates with poor prognosis. MDS globally affects all cellular subsets in bone marrow and in peripheral blood. We asked whether flow cytometry (FC) can separate individual subtypes of MDS from each other and from aplastic anemia (SAA) and whether in individual subtypes of childhood MDS can separate patients with and without monosomy 7.

Patients/analyzed parameters
In total we analyzed 94 children with centrally analyzed immunophenotype in the reference lab who were diagnosed and treated for MDS or SAA between 1998 and 2009. In total we analyzed 14 patients with refractory cytopenia, 37 patients with advanced forms of MDS (JMML 10, RAEB 25, CMML 2) and 43 patients with SAA. Monosomy 7/del(7q) was present in 17 patients (RC 6, JMML 3, RAEB 8). Analyzed parameters were as follows: B cells, CD10+CD19+, CD19+45dim/neg, CD19+34+, CD19/CD34 ratio, CD34+, CD117 cells, CD34+38dim/neg, CD3+, CD3+4+, CD3+8+, CD3+HLADR+.

Statistics
We analyzed all parameters using non parametric tests (Mann-Whitney, Kruskal Wallis) and principal component analysis (PCA).

Results
Principal component analysis of all analyzed patients together clearly separates advanced forms of MDS from RC and SAA, the most contributing factor being the number of CD34 and CD117+ cells. In non parametric statistics following factors significantly differ among MDS subtypes and SAA (Kruskal-Wallis): CD19, CD117, CD34, CD3, CD3+4+, CD8+ and CD3+HLADR+. RC and SAA patients are separated mainly by the number of B cells and the CD34:CD19 ratio. In addition, the following parameters differ between RC and SAA (Mann-Whitney): CD34, CD117 and CD3+HLADR+. Unlike the CD34:CD19 ratio, the number of CD19+34+ precursors does not differ between RC and SAA patients. Patients with monosomy 7 do not differ from the remaining patients when all MDS patients are analyzed together or separately in the respective subgroups (RC, non RC, JMML) by PCA or by non parametric statistics.

Conclusion
PCA separates advanced MDS forms from RC and SAA. Advanced forms of MDS are characterized by increased percentage of CD34+ and CD117+ cells compared to RC and SAA patients. The global reduction of B cell progenitor compartment is pronounced especially in non-JMML cases of MDS, whereas SAA patients typically present with isolated reduction of cells at early stages (CD19+34+) of B cell development. Patients with monosomy 7 cluster within the respective disease category, they do not form own cluster in PCA. Supported by MSMT VZ MSM0021620813, MZO 00064203 VZ FNM, MZO VFN2005, IGA NR/9531-3, NPV 2B06064.

Disclosures:
No relevant conflicts of interest to declare.