Identification of functionally primitive and immunophenotypically distinct subpopulations in secondary acute myeloid leukemia by mass cytometry
Identification of functionally primitive and immunophenotypically distinct subpopulations in secondary acute myeloid leukemia by mass cytometry
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DOI：
10.1002/cyto.b.21743
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发表时间：
2019-01-01
影响因子：
3.4
通讯作者：
Oh, Stephen T.
中科院分区：
文献类型：
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作者：
Bandyopadhyay, Shovik;Fowles, Jared S.;Oh, Stephen T.
背景背景：质谱流式细胞术（CyTOF）是在单细胞水平上分析细胞网络的强大工具。由于这种方法的高维性质，已经开发出分析算法来可视化和解释质谱流式数据。在这项研究中，我们将这些方法应用于一组继发性急性髓系白血病 (sAML) 患者。方法 我们利用质谱流式细胞术来研究 sAML 中血小板生成素介导的细胞内信号传导的定位和强度。使用 SPADE、viSNE 和 PhenoGraph 剖析细胞外和细胞内表型。结果健康对照表现出高度局部化的信号反应，很大程度上局限于造血干/祖细胞（HSPC）区室。相比之下，sAML 样本包含 HSPC 区外的亚群，其血小板生成素 (TPO) 敏感性与免疫表型定义的 HSPC 相当或更高。我们采用 PhenoGraph 进行无监督聚类来阐明这些异质样本中的不同亚群。鉴定出一个几乎完全由 Lin(-) CD61(+) CD34(-) CD38(-) CD45(low) 细胞组成的元簇。该亚群不易通过已建立的手动门控方法识别，并且通常在 TPO 刺激反应中表现出比 Lin(-) CD61(-) CD34(+) CD38(-) 细胞更高的 STAT 磷酸化。 Lin(-) CD61(+) CD34(-) CD38(-) CD45(low) 细胞在另外三名 sAML 患者中被鉴定出来，使用基于 PhenoGraph 结果的手动门控方法进行独立分析。每个患者都对 PhenoGraph 元簇表现出类似的 TPO 超敏反应。结论 该细胞亚群的鉴定凸显了 sAML 中手动门控的局限性。我们的研究证明了质谱流式分析仪通过利用无监督的高维分析来阐明高度异质性肿瘤中的稀有亚群的潜力。 (c) 2018年国际临床细胞计数学会
BackgroundBackground : Mass cytometry (CyTOF) is a powerful tool for analyzing cellular networks at the single cell level. Due to the high-dimensional nature of this approach, analysis algorithms have been developed to visualize and interpret mass cytometry data. In this study, we applied these approaches to a cohort of patients with secondary acute myeloid leukemia (sAML). Methods We utilized mass cytometry to interrogate localization and intensity of thrombopoietin-mediated intracellular signaling in sAML. Extracellular and intracellular phenotypes were dissected using SPADE, viSNE, and PhenoGraph. Results Healthy controls exhibited highly localized signaling responses largely restricted to the hematopoietic stem/progenitor cell (HSPC) compartment. In contrast, sAML samples contained subpopulations outside the HSPC compartment exhibiting thrombopoietin (TPO) sensitivity comparable to or greater than immunophenotypically defined HSPCs. We employed unsupervised clustering by PhenoGraph to elucidate distinct subpopulations within these heterogeneous samples. One metacluster composed almost exclusively of Lin(-) CD61(+) CD34(-) CD38(-) CD45(low) cells was identified. This subpopulation was not readily identified by established manual gating approaches, and generally exhibited greater STAT phosphorylation in response to TPO stimulation than did Lin(-) CD61(-) CD34(+) CD38(-) cells. Lin(-) CD61(+) CD34(-) CD38(-) CD45(low) cells were identified in three additional sAML patients analyzed independently using a manual gating approach based upon PhenoGraph results. Each patient exhibited a similar TPO hypersensitivity to the PhenoGraph metacluster. Conclusions The identification of this cellular subpopulation highlights the limitations of manual gating in sAML. Our study demonstrates the potential for mass cytometry to elucidate rare subpopulations in highly heterogeneous tumors by utilizing unsupervised high dimensional analysis. (c) 2018 International Clinical Cytometry Society