Overexpression of Nuclear Receptor 5A1 Induces and Maintains an Intermediate State of Conversion between Primed and Naive Pluripotency

Naive and primed human pluripotent stem cells (hPSCs) have provided useful insights into the regulation of pluripotency. However, the molecular mechanisms regulating naive conversion remain elusive. Here, we report intermediate naive conversion induced by overexpressing nuclear receptor 5A1 (NR5A1) in hPSCs. The cells displayed some naive features, such as clonogenicity, glycogen synthase kinase 3β, and mitogen-activated protein kinase (MAPK) independence, expression of naive-associated genes, and two activated X chromosomes, but lacked others, such as KLF17 expression, transforming growth factor β independence, and imprinted gene demethylation. Notably, NR5A1 negated MAPK activation by fibroblast growth factor 2, leading to cell-autonomous self-renewal independent of MAPK inhibition. These phenotypes may be associated with naive conversion, and were regulated by a DPPA2/4-dependent pathway that activates the selective expression of naive-associated genes. This study increases our understanding of the mechanisms regulating the conversion from primed to naive pluripotency. [Display omitted] •hPSCs in an intermediate state between primed and naive pluripotency are generated•Conversion to the state is induced by nuclear receptor 5A1 overexpression•The state displays phenotypic similarities and differences from primed and naive cells•DPPA2/4 regulate gene expression in conversion Human stem cells exist in two different states, primed and naive. However, the mechanisms regulating conversion between them remain unknown. Here, Hirofumi Suemori and colleagues report a stable intermediate conversion stage, induced by overexpressing nuclear receptor 5A1. This state is phenotypically distinct from primed and naive cells and provides a model to elucidate the molecular events involved in naive conversion.