Immunophenotypic characterization of leukemic stem cells in acute myeloid leukemia using single tube 10-colour panel by multiparametric flow cytometry: Deciphering the spectrum, complexity and immunophenotypic heterogeneity

IF 2.2 4区医学 Q3 HEMATOLOGY

International Journal of Laboratory Hematology Pub Date : 2024-03-08 DOI:10.1111/ijlh.14250

Nupur Das, Devasis Panda, Smeeta Gajendra, Ritu Gupta, Deepshi Thakral, Gurvinder Kaur, Aafreen Khan, Vivek Kumar Singh, Arushi Vemprala, Sameer Bakhshi, Rachna Seth, Ranjit Kumar Sahoo, Atul Sharma, Sandeep Rai, Vijay K. Prajapati, Saroj Singh

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引用次数: 0

Abstract

Introduction

Despite extensive research, comprehensive characterization of leukaemic stem cells (LSC) and information on their immunophenotypic differences from normal haematopoietic stem cells (HSC) is lacking. Herein, we attempted to unravel the immunophenotypic (IPT) characteristics and heterogeneity of LSC using multiparametric flow cytometry (MFC) and single-cell sequencing.

Materials and Methods

Bone marrow aspirate samples from patients with acute myeloid leukaemia (AML) were evaluated using MFC at diagnostic and post induction time points using a single tube-10-colour-panel containing LSC-associated antibodies CD123, CD45RA, CD44, CD33 and COMPOSITE (CLL-1, TIM-3, CD25, CD11b, CD22, CD7, CD56) with backbone markers that is, CD45, CD34, CD38, CD117, sCD3. Single-cell sequencing of the whole transcriptome was also done in a bone marrow sample.

Results

LSCs and HSCs were identified in 225/255 (88.2%) and 183/255 (71.6%) samples, respectively. Significantly higher expression was noted for COMPOSITE, CD45RA, CD123, CD33, and CD44 in LSCs than HSCs (p < 0.0001). On comparing the LSC specific antigen expressions between CD34+ (n = 184) and CD34- LSCs (n = 41), no difference was observed between the groups. More than one sub-population of LSC was demonstrated in 4.4% of cases, which further revealed high concordance between MFC and single cell transcriptomic analysis in one of the cases displaying three LSC subpopulations by both methods.

Conclusion

A single tube-10-colour MFC panel is proposed as an easy and reproducible tool to identify and discriminate LSCs from HSCs. LSCs display both inter- and intra-sample heterogeneity in terms of antigen expressions, which opens the facets for single cell molecular analysis to elucidate the role of subpopulations of LSCs in AML progression.

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通过多参数流式细胞术使用单管 10 色面板鉴定急性髓性白血病白血病干细胞的免疫表型特征：解密白血病的谱系、复杂性和免疫表型异质性。

简介尽管开展了大量研究，但仍缺乏白血病干细胞（LSC）的全面特征及其与正常造血干细胞（HSC）免疫表型差异的信息。在此，我们尝试使用多参数流式细胞术（MFC）和单细胞测序技术来揭示白血病干细胞的免疫表型（IPT）特征和异质性：在诊断时间点和诱导后时间点使用多参数流式细胞术评估急性髓性白血病（AML）患者的骨髓穿刺样本，使用单管-10色板，包含LSC相关抗体CD123、CD45RA、CD44、CD33和COMPOSITE（CLL-1、TIM-3、CD25、CD11b、CD22、CD7、CD56），以及骨干标记物CD45、CD34、CD38、CD117和sCD3。还对骨髓样本的全转录组进行了单细胞测序：结果：在 225/255 份样本（88.2%）和 183/255 份样本（71.6%）中分别发现了造血干细胞和造血干细胞。在低造血干细胞中，COMPOSITE、CD45RA、CD123、CD33 和 CD44 的表达量明显高于高造血干细胞（p）：建议使用单管-10 色 MFC 面板作为一种简便、可重复的工具来鉴别和区分 LSC 与 HSC。LSCs 在抗原表达方面显示出样本间和样本内的异质性，这为单细胞分子分析开辟了道路，有助于阐明 LSCs 亚群在急性髓细胞性白血病进展中的作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

International Journal of Laboratory Hematology 医学-血液学

CiteScore

4.50

自引率

6.70%

发文量

211

审稿时长

6-12 weeks

期刊介绍： The International Journal of Laboratory Hematology provides a forum for the communication of new developments, research topics and the practice of laboratory haematology. The journal publishes invited reviews, full length original articles, and correspondence. The International Journal of Laboratory Hematology is the official journal of the International Society for Laboratory Hematology, which addresses the following sub-disciplines: cellular analysis, flow cytometry, haemostasis and thrombosis, molecular diagnostics, haematology informatics, haemoglobinopathies, point of care testing, standards and guidelines. The journal was launched in 2006 as the successor to Clinical and Laboratory Hematology, which was first published in 1979. An active and positive editorial policy ensures that work of a high scientific standard is reported, in order to bridge the gap between practical and academic aspects of laboratory haematology.