Guangzhou Institute of Biology reveals the molecular mechanism of the transformation process of mouse pluripotent stem cells from the initial state to the original state
The transformation of cell fate is determined by precise and complex signal regulation and epigenetic regulation, and the study of its molecular mechanism is one of the core problems that need to be solved urgently in the development of cell biology. Studies have shown that mouse pluripotent stem cells have two different states in signal maintenance and function, namely, primitive Epiblast stem cells, EpiSCs) and naive Embryonic stem cells, ESCs). In vitro, these two kinds of pluripotent stem cells can transform each other. Under the stimulation of ActivinA and bFGF signals, naive ESCs can differentiate into primed EpiSCs. On the contrary, by overexpressing a single naive pluripotent gene, such as Esrrb, Klf2, Nr5a2, etc., the initial to naive transition (PNT) can also be realized. Studying the mechanism of cell fate change in mouse PNT process can provide reference for isolating or inducing naive pluripotent stem cells from different species, but the main molecular mechanism of cell fate change in mouse PNT is still unclear.
On May 11th, Pei Duanqing’s research group and Liu Jing’s research group of Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences published a research paper entitled BMP4 resets mouse epi blast stem cells to naive proliferation through zbtb7a/b-mediated chromatin remolding online in the journal Nature Cell Biology. In this study, we first found that BMP4 can induce PNT in mice, and established an efficient PNT induction system with clear components. Subsequently, high-throughput sequencing techniques such as ATAC-seq were used to reveal the specific molecular mechanism of mouse PNT process.
At first, researchers found that growth factor BMP4 can induce PNT in mice. With the help of visual reporting system and screening of compound library, it was found that EZH2 inhibitor EPZ6438 and DOT1L inhibitor EPZ5676 can significantly improve the induction efficiency, and then a mouse PNT induction system with a clear chemical composition with an induction efficiency of 80% within 8 days was established.
On the basis of this platform, researchers combined bioinformatics analysis based on chromatin transposase accessibility sequencing (ATAC-Seq). The dynamic changes of chromatin accessibility in the process of PNT induced by small chemical molecules in mice were described, and it was found that the most critical factor in this system, BMP4, mediated the progress of PNT by regulating chromatin accessibility, on the one hand, inhibiting the opening of differentiation-related sites, on the other hand, promoting the opening of naive pluripotent gene sites. By analyzing the gene sites where BMP4 opens and activates expression, the researchers found that the new downstream targets of BMP4, Zbtb7a and Zbtb7b, regulate the function of PNT by influencing chromatin remodeling. Furthermore, chromatin immunoprecipitation (Chip) proved that Zbtb7a directly binds to and activates the expression of naive pluripotent genes such as Esrrb, Klf2, Nr5a2, thus regulating the occurrence of PNT.
To sum up, this study established an efficient technical platform for chemical induction of mouse PNT, revealed the molecular mechanism and found new targets from the perspective of chromatin accessibility, and made new discoveries in the study of cell fate transformation mechanism, which provided a new understanding and breakthrough point for the study of chemical regulation of cell fate transformation, and also provided a very meaningful reference direction for other research systems, such as the acquisition of naive pluripotent stem cells in human system.
Researchers Pei Duanqing and Liu Jing are the corresponding authors of this paper. Doctoral students Yu Shengyong and Zhou Chunhua, Ph.D. students Cao Shangtao and He Jiangping are the co-first authors of this paper.
Paper link
Cell fate transition process, chromatin changes and intervening regulatory proteins