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Profiling myelodysplastic syndromes by mass cytometry demonstrates abnormal progenitor cell phenotype and differentiation.
Cytometry Part B: Clinical Cytometry ( IF 2.3 ) Pub Date : 2020-01-09 , DOI: 10.1002/cyto.b.21860
Gregory K Behbehani 1, 2, 3 , Rachel Finck 1 , Nikolay Samusik 1 , Kunju Sridhar 2 , Wendy J Fantl 4 , Peter L Greenberg 2, 3 , Garry P Nolan 1, 3, 4
Affiliation  

BACKGROUND We sought to enhance the cytometric analysis of myelodysplastic syndromes (MDS) by performing a pilot study of a single cell mass cytometry (MCM) assay to more comprehensively analyze patterns of surface marker expression in patients with MDS. METHODS Twenty-three MDS and five healthy donor bone marrow samples were studied using a 34-parameter mass cytometry panel utilizing barcoding and internal reference standards. The resulting data were analyzed by both traditional gating and high-dimensional clustering. RESULTS This high-dimensional assay provided three major benefits relative to traditional cytometry approaches: First, MCM enabled detection of aberrant surface maker at high resolution, detecting aberrancies in 27/31 surface markers, encompassing almost every previously reported MDS surface marker aberrancy. Additionally, three previously unrecognized aberrancies in MDS were detected in multiple samples at least one developmental stage: increased CD321 and CD99; and decreased CD47. Second, analysis of the stem and progenitor cell compartment (HSPCs), demonstrated aberrant expression in 21 of the 23 MDS samples, which were not detected in three samples from patients with idiopathic cytopenia of undetermined significance. These immunophenotypically abnormal HSPCs were also the single most significant distinguishing feature between clinical risk groups. Third, unsupervised clustering of high-parameter MCM data allowed identification of abnormal differentiation patterns associated with immunophenotypically aberrant myeloid cells similar to myeloid derived suppressor cells. CONCLUSIONS These results demonstrate that high-parameter cytometry methods that enable simultaneous analysis of all bone marrow cell types could enhance the diagnostic utility of immunophenotypic analysis in MDS.

中文翻译:

通过大规模细胞术分析骨髓增生异常综合征显示异常的祖细胞表型和分化。

背景 我们试图通过对单细胞大规模细胞计数 (MCM) 测定法进行初步研究以更全面地分析 MDS 患者的表面标志物表达模式,从而增强骨髓增生异常综合征 (MDS) 的细胞计数分析。方法 23 个 MDS 和 5 个健康供体骨髓样本使用条形码和内部参考标准的 34 参数质谱仪组进行研究。通过传统的门控和高维聚类分析得到的数据。结果与传统细胞计数方法相比,这种高维分析提供了三个主要优势:首先,MCM 能够以高分辨率检测异常表面标记物,检测 27/31 表面标记物的异常,包括几乎所有先前报道的 MDS 表面标记物异常。此外,在至少一个发育阶段的多个样本中检测到三种以前未被识别的 MDS 异常:CD321 和 CD99 增加;并减少 CD47。其次,对干细胞和祖细胞区室 (HSPC) 的分析表明,在 23 个 MDS 样本中的 21 个中存在异常表达,而在来自具有未确定意义的特发性血细胞减少症患者的三个样本中未检测到这些异常表达。这些免疫表型异常的 HSPC 也是临床风险组之间最显着的区别特征。第三,高参数 MCM 数据的无监督聚类允许识别与类似于髓源抑制细胞的免疫表型异常髓细胞相关的异常分化模式。
更新日期:2020-01-09
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