Integrative epigenomic and transcriptomic analysis reveals the requirement of JUNB for hematopoietic fate induction

To advance our understanding of the dynamics of the gene regulation during each step of in vitro HSPC, we surveyed global gene expression, chromatin accessibility (ATAC-seq), chromatin immunoprecipitation sequencing (ChIP-seq) for active H3K4me3 and repressive H3K27me3 histone mark, from hPSCs and purified intermediates across HSPC differentiation. Methods: ATAC-seq, bulk RNA-seq, scRNA-seq, ChIP-seq, scATAC-seq, Cut & Tag Overall design: Bulk RNA-seq, ATAC-seq, H3K4me3 and H3K27me3 ChIP-seq were performed with hPSCs and hPSC derived VMEs, EPCs and HPCs . Besides, we performed scATAC-seq and scRNA-seq to investigate the transcriptomic signature of HEC generated in in vitro system.

为加深对体外造血干细胞与祖细胞（Hematopoietic Stem and Progenitor Cells，HSPC）各分化阶段基因调控动态变化的理解，我们针对人类多能干细胞（human pluripotent stem cells，hPSCs）以及造血干细胞与祖细胞分化过程中纯化得到的各中间阶段样本，开展了全基因表达谱检测、染色质开放状态分析（转座酶可及性染色质测序，ATAC-seq），以及针对活化型组蛋白H3赖氨酸4三甲基化（H3K4me3）与抑制型组蛋白H3赖氨酸27三甲基化（H3K27me3）的染色质免疫共沉淀测序（ChIP-seq）分析。
实验方法：涵盖ATAC-seq、批量RNA测序（bulk RNA-seq）、单细胞RNA测序（scRNA-seq）、ChIP-seq、单细胞ATAC-seq（scATAC-seq）及Cut & Tag技术。
实验设计：我们对人类多能干细胞（hPSCs）及其分化获得的VMEs、内皮祖细胞（EPCs）、造血祖细胞（HPCs）开展了批量RNA-seq、ATAC-seq、H3K4me3及H3K27me3 ChIP-seq实验。此外，我们通过scATAC-seq与scRNA-seq，探究了体外体系中生成的造血内皮细胞（HECs）的转录组特征。