生物
单倍率不足
祖细胞
造血
干细胞
癌症研究
转录因子
分子生物学
细胞生物学
运行x1
遗传学
表型
基因
作者
Brian Estevez,Sara Borst,Danuta Jarocha,Varun Sudunagunta,Michael V. Gonzalez,James Garifallou,Hákon Hákonarson,Peng Gao,Kai Tan,Paul P. Liu,Sumedha Bagga,Nicholas Holdreith,Wei Tong,Nancy A. Speck,Deborah L. French,Paul Gadue,Mortimer Poncz
出处
期刊:Blood
[American Society of Hematology]
日期:2021-05-13
卷期号:137 (19): 2662-2675
被引量:15
标识
DOI:10.1182/blood.2020006389
摘要
Abstract Patients with familial platelet disorder with a predisposition to myeloid malignancy (FPDMM) harbor germline monoallelic mutations in a key hematopoietic transcription factor, RUNX-1. Previous studies of FPDMM have focused on megakaryocyte (Mk) differentiation and platelet production and signaling. However, the effects of RUNX-1 haploinsufficiency on hematopoietic progenitor cells (HPCs) and subsequent megakaryopoiesis remains incomplete. We studied induced pluripotent stem cell (iPSC)–derived HPCs (iHPCs) and Mks (iMks) from both patient-derived lines and a wild-type (WT) line modified to be RUNX-1 haploinsufficient (RUNX-1+/−), each compared with their isogenic WT control. All RUNX-1+/− lines showed decreased iMk yield and depletion of an Mk-biased iHPC subpopulation. To investigate global and local gene expression changes underlying this iHPC shift, single-cell RNA sequencing was performed on sorted FPDMM and control iHPCs. We defined several cell subpopulations in the Mk-biased iHPCs. Analyses of gene sets upregulated in FPDMM iHPCs indicated enrichment for response to stress, regulation of signal transduction, and immune signaling-related gene sets. Immunoblot analyses in FPDMM iMks were consistent with these findings, but also identified augmented baseline c-Jun N-terminal kinase (JNK) phosphorylation, known to be activated by transforming growth factor-β1 (TGF-β1) and cellular stressors. These findings were confirmed in adult human CD34+-derived stem and progenitor cells (HSPCs) transduced with lentiviral RUNX1 short hairpin RNA to mimic RUNX-1+/−. In both iHPCs and CD34+-derived HSPCs, targeted inhibitors of JNK and TGF-β1 pathways corrected the megakaryopoietic defect. We propose that such intervention may correct the thrombocytopenia in patients with FPDMM.
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