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Elucidating the role of megakaryocytes as immune cells using iPSC-derived megakaryocyte progenitor models

使用 iPSC 衍生的巨核细胞祖模型阐明巨核细胞作为免疫细胞的作用

基本信息

批准号：
22K15124
负责人：
Chen SiJing
金额：
$ 2.91万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Early-Career Scientists
财政年份：
2022
资助国家：
日本
起止时间：
2022-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-22K15124/
关键词：
iPS 巨核球免疫 Megakaryocyte iPSC immune system

项目摘要

Recent evidence indicates that human megakaryocytes (MKs) represent a heterogeneous population, which includes an immune subset that is genetically programmed. However, the ontogeny of this heterogeneity in human MKs is not well understood. In this study, we sought to investigate the regulation of immune-biased MKs utilizing an immortalized MK cell line (imMKCLs), which is derived from human-induced pluripotent stem cells (iPSCs) and serves as a source of iPSC-derived platelets (iPSC-PLTs) used in clinical trials.In 2022, we conducted a series of studies aimed at characterizing the immune properties of imMKCLs and elucidating the key regulators involved in immune-skewed imMKCLs. Specifically, we investigated the expression of several immune-related surface markers and found ICAM-1 and CD11b were expressed on imMKCLs. To identify potential factors involved in the lineage determination of immune-biased transcriptional signatures during imMKCL development, we conducted single-cell RNA-seq analysis using imMKCLs. Through gain-of-function and lentiviral-mediated forced expression studies, we identified let-7a-5p and its downstream target gene X as potential key regulators involved in immune-skewed imMKCL development. Unexpectedly, we discovered that the dysregulated immune properties/subsets of imMKCLs were closely linked with deficient proliferation and subsequently impaired PLT production.

最近的证据表明，人类巨核细胞（MK）代表了一个异质性群体，其中包括一个免疫子集，是遗传编程。然而，这种异质性在人类MK的个体发育还没有得到很好的理解。在这项研究中，我们试图利用永生化MK细胞系（imMKCL）来研究免疫偏向性MK的调节，该细胞系源自人诱导多能干细胞（iPSC），并作为iPSC衍生血小板的来源（iPSC-PLT）用于临床试验。在2022年，我们进行了一系列旨在表征imMKCL的免疫特性和阐明免疫偏斜imMKCL中涉及的关键调节剂的研究。具体而言，我们研究了几种免疫相关表面标志物的表达，发现ICAM-1和CD 11b在imMKCL上表达。为了鉴定在imMKCL开发过程中参与免疫偏倚转录特征谱系确定的潜在因素，我们使用imMKCL进行了单细胞RNA-seq分析。通过功能获得和慢病毒介导的强制表达研究，我们确定let-7a-5 p及其下游靶基因X是参与免疫偏斜imMKCL发展的潜在关键调节因子。出乎意料的是，我们发现imMKCL的免疫特性/亚群失调与增殖缺陷和随后受损的PLT产生密切相关。