DMRT1 regulates human germline commitment.
Germline commitment following primordial germ cell (PGC) specification during early human development establishes an epigenetic programme and competence for gametogenesis. Here we follow the progression of nascent PGC-like cells derived from human embryonic stem cells in vitro. We show that switching from BMP signalling for PGC specification to Activin A and retinoic acid resulted in DMRT1 and CDH5 expression, the indicators of migratory PGCs in vivo. Moreover, the induction of DMRT1 and SOX17 in PGC-like cells promoted epigenetic resetting with striking global enrichment of 5-hydroxymethylcytosine and locus-specific loss of 5-methylcytosine at DMRT1 binding sites and the expression of DAZL representing DNA methylation-sensitive genes, a hallmark of the germline commitment programme. We provide insight into the unique role of DMRT1 in germline development for advances in human germ cell biology and in vitro gametogenesis.
Acknowledgements: We thank J. Cerviera, M. Jarana and C. Bradford for assistance with flow cytometry and cell sorting; R. Butler for generating a program for immunofluorescence image analysis; S. Kim for critical suggestions for the study and technical assistance; the Austin Smith lab for providing Shef-6 cell line and technical discussions; the Rick Livesey lab for supporting tissue culture; H. Saito, M. Gu, N. Merleau-Ponty, E. Sen and C. Lee for technical assistance; former and current Surani lab members and Gurdon Institute members for input, discussion and support for the research; and M. Suematsu, the Department of Biochemistry & Integrative Medical Biology, School of Medicine, Keio University and C. le Sage for critical support. The research was supported by an MRC research grant (RG85305) and Rosetrees Trust to M.A.S. and N.I., by a Wellcome Investigator Awards in Science (209475/Z/17/Z and 096738/Z/11/Z) and a BBSRC research grant (G103986) to M.A.S., by JSPS KAKENHI Grant (JP23H03047) to N.I. and by a core grant to the Gurdon Institute by Wellcome and Cancer Research UK. S.-M.L. is supported by the Brain Pool programme funded by the Ministry of Science and ICT through the National Research Foundation of Korea (2022H1D3A2A02063272). T.K. is supported by AMED (JP22bm1123008). The C.-X.S. lab is supported by the Ludwig Institute for Cancer Research, Cancer Research UK (C63763/A26394 and C63763/A27122), and National Institute for Health Research (NIHR) Oxford Biomedical Research Centre (BRC). H.X. is supported by China Scholarship Council. M.I. is supported by the Nakajima Foundation. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health.
Funder: Brain Pool program funded by the Ministry of Science and ICT through the National Research Foundation of Korea (2022H1D3A2A02063272)
Funder: Wellcome Sanger core funding
Funder: China Scholarship Council
Funder: Ludwig Institute for Cancer Research, Cancer Research UK (C63763/A26394 and C63763/A27122), and National Institute for Health Research (NIHR) Oxford Biomedical Research Centre (BRC)
Funder: Nakajima Foundation
Funder: AMED (JP22bm1123008)
Wellcome Trust (096738/Z/11/Z)