Analysis of Nr1h2 and H3K27ac genomic binding targets in ESC and EPSC under Nr1h2 activation

ChIP-seq for Nr1h2 in NrESC, ESC. H3K27ac ChIP-seq in NrESC, ESC, EPSC, EPSCT09 Embryonic stem cells upon extrinsic induction could self-assemble into blastocyst-like structures. However, the intrinsic regulation of such blastoid forming potential remain to be addressed. We discover that the activity of nuclear receptor subfamily 1, group H, member 2 (Nr1h2) in expanded potential stem cell (EPSC) positively correlates with blastoid efficiency and quality. In addition, Nr1h2 agonist, T0901317, improves natural blastocyst development. Surprisingly, Nr1h2-activated ESC (NrESC) is rewired towards a distinct pluripotency state that is capable of self-organizing into blastoids and contribute to embryonic and extraembryonic lineage. We hypothesize that Nr1h2 activation broadly affects the transcriptome landscape by regulating chromatin accessibility and epigenome to rewire ESC state. Indeed, from Nr1h2 and H3K27Ac ChIP-seq, we observe that annotated genes with upregulated Nr1h2 binding as well as H3K27Ac marks in NrESC or EPSC treated with T09 are enriched in processes such as lipid metabolism, pluripotency maintenance and trophectoderm maturation, which are known to be essential to blastocyst development. In summary, our study demonstrates a novel Nr1h2-centric regulation of expanded pluripotency.