Bachas Costa, Duetz Carolien, van Spronsen Margot F, Verhoeff Jan, Garcia Vallejo Juan J, Jansen Joop H, Cloos Jacqueline, Westers Theresia M, van de Loosdrecht Arjan A
Department of Hematology, Cancer Center Amsterdam, Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
Department of Molecular Cell Biology and Immunology, Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
Cytometry B Clin Cytom. 2023 Mar;104(2):128-140. doi: 10.1002/cyto.b.22066. Epub 2022 Mar 15.
Myelodysplastic syndromes (MDS) at risk of transformation to acute myeloid leukemia (AML) are difficult to identify. The bone marrows of MDS patients harbor specific hematopoietic stem and progenitor cell (HSPC) abnormalities that may be associated with sub-types and risk-groups. Leukemia-associated characteristics of such cells may identify MDS patients at risk of progression to AML and provide insight in the pathobiology of MDS.
Bone marrow samples from healthy donors (n = 10), low risk (n = 12) and high risk (n = 13) MDS patients were collected, in addition, AML samples for 5 out of 6 MDS patients that progressed. Mass cytometry was applied to assess expression of stem cell subset and leukemia-associated immunophenotype markers.
We analyzed the data using FlowSOM to cluster cells with similar expression of 10 commonly used stem cell markers. Metaclusters (n = 20) of these clusters represented populations of cells with a related phenotype, largely resembling known stem cell subsets. Within specific subsets, intra-cellular expression levels of pCREB, IkBα, or pS6 differed significantly between healthy bone marrow (HBM) and MDS or consecutive secondary AML samples. CD34, CD44, and CD49f expression was significantly increased in high risk MDS and AML-associated metaclusters. We identified MDS/sAML cells with aberrant phenotypes when compared to HBM. Such cells were observed in clusters of both primary MDS and secondary AML samples.
High-dimensional mass cytometry and computational data analyses enabled characterization of HSPC subsets in MDS and identification of leukemia stem cell populations based on their immunophenotype. Stem cells in MDS that display leukemia-associated features may predict the risk of developing AML.
难以识别有转化为急性髓系白血病（AML）风险的骨髓增生异常综合征（MDS）。MDS患者的骨髓存在特定的造血干细胞和祖细胞（HSPC）异常，这可能与亚型和风险组相关。此类细胞的白血病相关特征可能有助于识别有进展为AML风险的MDS患者，并为MDS的病理生物学提供见解。
收集了健康供者（n = 10）、低风险（n = 12）和高风险（n = 13）MDS患者的骨髓样本，此外，还收集了6例进展为AML的MDS患者中的5例的AML样本。采用质谱流式细胞术评估干细胞亚群和白血病相关免疫表型标志物的表达。
我们使用FlowSOM分析数据，以对10种常用干细胞标志物表达相似的细胞进行聚类。这些聚类的元聚类（n = 20）代表具有相关表型的细胞群体，在很大程度上类似于已知的干细胞亚群。在特定亚群中，健康骨髓（HBM）与MDS或后续继发性AML样本之间，pCREB、IkBα或pS6的细胞内表达水平存在显著差异。高风险MDS和AML相关元聚类中CD34、CD44和CD49f的表达显著增加。与HBM相比，我们鉴定出具有异常表型的MDS/sAML细胞。在原发性MDS和继发性AML样本的聚类中均观察到此类细胞。
高维质谱流式细胞术和计算数据分析能够对MDS中的HSPC亚群进行表征，并基于免疫表型识别白血病干细胞群体。显示白血病相关特征的MDS干细胞可能预测发生AML的风险。
