In vitro reconstitution of epigenetic reprogramming in the human germ line

• 成果类型: Article
• 署名作者: Murase, Yusuke; Yokogawa, Ryuta; Yabuta, Yukihiro; Nagano, Masahiro; Katou, Yoshitaka; Mizuyama, Manami; Kitamura, Ayaka; Puangsricharoen, Pimpitcha; Yamashiro, Chika; Hu, Bo; Mizuta, Ken; Tsujimura, Taro; Yamamoto, Takuya; Ogata, Kosuke; Ishihama, Yasushi; Saitou, Mitinori
• 署名单位: Kyoto University; Kyoto University; Kyoto University; RIKEN; Kyoto University
• 刊物名称: Nature
• ISSN/ISSBN: 0028-5810
• DOI: 10.1038/s41586-024-07526-6
• 发表日期: 2024-07-04
• 关键词: pluripotent stem-cells; dna methylation; tet proteins; induction; specification; fate; gene; identification; trophoblast; mechanism

摘要

Epigenetic reprogramming resets parental epigenetic memories and differentiates primordial germ cells (PGCs) into mitotic pro-spermatogonia or oogonia. This process ensures sexually dimorphic germ cell development for totipotency(1). In vitro reconstitution of epigenetic reprogramming in humans remains a fundamental challenge. Here we establish a strategy for inducing epigenetic reprogramming and differentiation of pluripotent stem-cell-derived human PGC-like cells (hPGCLCs) into mitotic pro-spermatogonia or oogonia, coupled with their extensive amplification (about >10(10)-fold). Bone morphogenetic protein (BMP) signalling is a key driver of these processes. BMP-driven hPGCLC differentiation involves attenuation of the MAPK (ERK) pathway and both de novo and maintenance DNA methyltransferase activities, which probably promote replication-coupled, passive DNA demethylation. hPGCLCs deficient in TET1, an active DNA demethylase abundant in human germ cells(2,3), differentiate into extraembryonic cells, including amnion, with de-repression of key genes that bear bivalent promoters. These cells fail to fully activate genes vital for spermatogenesis and oogenesis, and their promoters remain methylated. Our study provides a framework for epigenetic reprogramming in humans and an important advance in human biology. Through the generation of abundant mitotic pro-spermatogonia and oogonia-like cells, our results also represent a milestone for human in vitro gametogenesis research and its potential translation into reproductive medicine.