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CHAF1A通过激活NF-Y转录因子活性维持造血干细胞干性

刘进¹, 萧梦杜², 游婉玲³, 盛岳¹, 汪虎^2,*

¹中南大学湘雅二医院血液科，长沙 410011；²杭州师范大学基础医学院，浙江省医学表观遗传学重点实验室，杭州 311121；³暨南大学教育部再生医学重点实验室，广州 510632

摘要

本研究旨在探究染色质组装因子1亚基CHAF1A对成年小鼠造血干细胞(hematopoietic stem cells, HSCs)功能的调控作用及潜在分子机制。通过构建造血系统特异性Chaf1a 基因敲除小鼠模型，结合流式细胞术，转录组测序(RNA-seq)、ATAC-seq 及HINT-ATAC技术进行多维分析。结果显示，CHAF1A缺失小鼠外周血血小板数量显著减少，骨髓中造血干祖细胞即LSK (lineage- Sca-1+ c-Kit+)细胞和HSCs 数目显著减少，而造血祖细胞[粒细胞-单核细胞祖细胞(granulocyte-monocyteprogenitors, GMP)、巨核细胞/红系祖细胞(megakaryocyte-erythroid progenitors, MEP)、共同髓系祖细胞(common myeloidprogenitors, CMP)]数目无显著差异，细胞周期及凋亡比例未见显著异常。转录组测序显示，CHAF1A缺失HSCs中有904 个基因显著上调、781 个基因显著下调；基因集富集分析证实，干性相关基因在野生型HSCs 中富集，分化相关基因富集于CHAF1A缺失组。ATAC-seq 结果显示，CHAF1A缺失组HSCs转录起始位点区域染色质开放程度降低。HINT-ATAC分析进一步表明，NF-Y家族转录因子活性在野生型HSCs中更高，而CEBPA、CEBPD活性在CHAF1A缺失HSCs中显著升高。以上结果表明，CHAF1A通过增强NF-Y 家族转录因子活性维持HSCs 干性，并抑制CEBP家族转录因子功能以阻滞分化，对成年小鼠HSCs维持至关重要，其机制涉及染色质可及性与转录因子活性的调控。

关键词： Chaf1a基因; 造血干细胞; 染色质可及性; 转录因子NF-Y; 录因子CEBP

CHAF1A sustains hematopoietic stem cell stemness by enhancing the activity of the NF-Y transcription factor family

LIU Jin¹, XIAO Meng-Du², YOU Wan-Ling³, SHENG Yue¹, WANG Hu^2,*

¹Department of Hematology, the Second Xiangya Hospital, Central South University, Changsha 410011, China；²Zhejiang Key Laboratory of Medical Epigenetics, School of Basic Medical Sciences, Hangzhou Normal University, Hangzhou 311121, China；³Key Laboratory of Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou 510632, China

Abstract

This study aimed to investigate the regulatory role and underlying molecular mechanisms of chromatin assembly factor 1 subunit CHAF1A in the function of hematopoietic stem cells (HSCs) in adult mice. A hematopoietic-specific Chaf1a knockout mouse model was established, and multiple approaches including flow cytometry, RNA-seq, ATAC-seq, and HINT-ATAC were integrated for multidimensional systematic analysis. CHAF1A depletion led to a pronounced reduction in peripheral blood platelet count. In the bone marrow, the number of LSK (lineage- Sca-1+ c-Kit+) cells and HSCs was markedly decreased, while no significant difference was observed in the number of hematopoietic progenitor cells (common myeloid progenitors, megakaryocyte-erythroid progenitors and granulocyte-monocyte progenitors) in the CHAF1A-deficient group. Additionally, no significant differences were observed in HSC cell cycle distribution or apoptosis. RNA-seq identified 904 upregulated and 781 downregulated genes in CHAF1A-deficient HSCs. Gene set enrichment analysis (GSEA) showed that stemness-related genes were significantly enriched in wild-type HSCs, while differentiation-related genes were significantly enriched in CHAF1A-deficient HSCs. ATAC-seq revealed reduced chromatin accessibility at the transcription start site (TSS) regions of HSCs in the CHAF1A-deficient group. HINT-ATAC analysis further showed that the activity of NF-Y family transcription factors was significantly higher in wild-type HSCs than in CHAF1A-deficient HSCs, while the activities of CEBPA and CEBPD were significantly higher in CHAF1A-deficient HSCs. Collectively, these results suggest that CHAF1A is essential for the maintenance of adult mouse HSCs. Specifically, CHAF1A enhances the activity of NF-Y family transcription factors to sustain HSC stemness and inhibits the function of CEBP family transcription factors to block HSC differentiation. Its regulatory mechanism involves the coordinated regulation of chromatin accessibility and transcription factor activity.

Key words: Chaf1a; hematopoietic stem cells; chromatin accessibility; transcription factor NF-Y; transcription factor CEBP

收稿日期：　　录用日期：

通讯作者：汪虎　　E-mail:

DOI: 10.13294/j.aps.2026.0069

引用本文：

刘进, 萧梦杜, 游婉玲, 盛岳, 汪虎. CHAF1A通过激活NF-Y转录因子活性维持造血干细胞干性[J]. 生理学报 2026; 78 (4): 931-942.

LIU Jin, XIAO Meng-Du, YOU Wan-Ling, SHENG Yue, WANG Hu. CHAF1A sustains hematopoietic stem cell stemness by enhancing the activity of the NF-Y transcription factor family. Acta Physiol Sin 2026; 78 (4): 931-942 (in Chinese with English abstract).