Congratulations | Najie research group of Medical College revealed the function of junB in the process of differentiation of artificial blood progenitor cells in vitro through multi group analysis

Najie research group of medical college has been engaged in hPSC In vitro differentiation into cardiovascular and hematopoietic lineages .2022 year 6 month 6 Japan , The research group is in 《 natural • Communications 》(Nature Communications) The magazine published the title “ Chromatin dynamics and transcriptomic analysis revealed JUNB An essential role in the generation of hematopoietic progenitor cells ‎(Integrative epigenomic and transcriptomic analysis reveals the requirement of JUNB for hematopoietic fate induction ) The article .（ Reprinted from ： Official account of Medical College of Tsinghua University ）

Hematopoietic stem progenitor cells （hematopoietic stem progenitor cell, HSPC） Present in adult bone marrow , Have long-term self-renewal ability , At the same time, it can differentiate into all types of blood cells and immune cells , Therefore, it has important value in disease treatment . Human embryos HSPC The study of single cell transcriptome map shows a unique dynamic change law (Zeng et al., 2019), But limited by materials and Technology , Human embryo HSPC The open state of chromatin during development 、 The epigenetic map of important histone modifications and the mechanism of gene expression regulation have not been revealed . Human pluripotent stem cells （human pluripotent stem cell, hPSC） To hematopoietic progenitor cells （hematopoietic progenitor cell, HPC） The system of differentiation is exploration HSPC Epigenetic regulation in the generative process provides a platform for experimental operation .

chart 1. HPC Study on gene expression regulation during differentiation in vitro

Researchers use hPSC towards HPC Phased differentiation system , Purified vascular mesoderm (vascular mesoderm cells, VME), Endothelial precursor cells (endothelial progenitor cell, EPC) And hematopoietic progenitor cells (HPC), The transcriptome of each differentiation stage was obtained 、 Open chromatin 、H3K4me3 and H3K27me3 Histone modification data . Through data integration and analysis , Based on the open chromatin map, the researchers inferred the transcription factors that may function at each stage , The divalent modification of histone (bivalent) And the epigenetic modification patterns of marker genes .

HSPC From a special group of endothelial cells - Hemopoietic endothelial cell （hemogenic endothelial cell, HEC）.HEC The characteristics of have a direct impact on HSPC Generation efficiency and whether it has the ability of transplantation and reconstruction (Guibentif et al., 2017; Park et al., 2018). The researchers used single cells RNA-seq and ATAC-seq technology , The differentiation in vitro was identified HEC Groups , And combine it with the HEC Transcriptome differences were compared . Results found , Produced in vitro HEC Although it has strong proliferative ability , But the gene expression of arterial endothelium is obviously insufficient . and HEC The characteristics of arterial endothelium are right HSPC The generation and function of (Zeng et al., 2019), therefore , In vitro differentiation system , enhance HEC The characteristics of arterial endothelium may help to improve HPC Differentiation potential .

after , be based on ATAC-seq Results of enrichment analysis of transcription factors and dynamic changes of histone modification , The researchers found that in hPSC towards HPC In the process of differentiation , transcription factor JUNB The dynamic changes of gene expression and the binding characteristics in the genome are related to MYB、RUNX1 And other important hematopoietic regulatory transcription factors . The researchers used JUNB Knocked out hPSC The differentiation of HPC, Find out JUNB The absence of significantly inhibited HEC and HPC Generation ,JUNB The complement experiment further proves JUNB stay HPC An important role in the process of differentiation . In terms of mechanism ,JUNB The existence of makes ERG、GATA2 and RUNX1 And the chromatin of hematopoietic lineage related factors showed an open state , promote HEC The specialization and orientation of HPC The transformation of ; in addition ,JUNB Can be combined directly RUNX1、CD44 Isoendothelium - Hematopoietic transformation (endothelial-to-hematopoietic transition, EHT) Promoter region of regulatory factors , Regulate gene expression and EHT Happen . thus JUNB yes hPSC The differentiation of HPC A necessary factor in the process .

All in all , This study reveals that humans HPC Epigenetic regulation mechanism during differentiation , Found out HEC and HPC New regulatory factors in the formation process JUNB. meanwhile , Inside and outside HEC The comparative analysis of single cell transcriptome is to obtain high-quality in vitro more efficiently HPC Provides important clues .

Associate Professor Na Jie is the corresponding author of this paper , Chen Xia, postdoctoral fellow of Tsinghua University Medical School 、 Doctoral students wangpeiliang and Qiuhui are the co first authors of this paper . Dr. duanfuyu from Guangzhou women and children medical center 、 Dr. guojianying from the Department of Obstetrics and Gynecology of the third Peking Medical College and the Chinese Academy of Medical Sciences / Professor Huang Yue of Peking Union Medical College provided important help for this study . Dr. zhuyonglin from Najie Laboratory 、 Doctoral student zhangxingwu 、 Zhangyaxuan and dingshuangyuan also made important contributions to this study . This study is sponsored by the national key project of stem cell and transformation research 2019YFA0110001、2017YFA0102802, Project of National Natural Science Foundation of China 31970819、91740115、31771108、32000610, Spring breeze fund of Tsinghua University 2021Z99CFY033, tsinghua - Peking University Union Center for life sciences 、 Protein research technology center 、 The advanced and sophisticated innovation center of structural biology, etc. provide funds and test instrument support .

Link to the original text

https://www.nature.com/articles/s41467-022-30789-4

reference

1、Guibentif, C., Ronn, R.E., Boiers, C., Lang, S., Saxena, S., Soneji, S., Enver, T., Karlsson, G., and Woods, N.B. (2017). Single-Cell Analysis Identifies Distinct Stages of Human Endothelial-to-Hematopoietic Transition. Cell Reports 19, 10-19. 10.1016/j.celrep.2017.03.023.

2、Park, M.A., Kumar, A., Jung, H.S., Uenishi, G., Moskvin, O.V., Thomson, J.A., and Slukvin, I.I. (2018). Activation of the Arterial Program Drives Development of Definitive Hemogenic Endothelium with Lymphoid Potential. Cell Reports 23, 2467-2481. 10.1016/j.celrep.2018.04.092.

3、Zeng, Y., He, J., Bai, Z., Li, Z., Gong, Y., Liu, C., Ni, Y., Du, J., Ma, C., Bian, L., et al. (2019). Tracing the first hematopoietic stem cell generation in human embryo by single-cell RNA sequencing. Cell Res 29, 881-894. 10.1038/s41422-019-0228-6.