a-Ketoglutarate promotes trophectoderm induction and maturation from naive human embryonic stem cells [scRNA-Seq 2D]

Development and lineage choice are driven by interconnected transcriptional, epigenetic and metabolic changes. Specific metabolites, such as a-ketoglutarate (aKG), function as signalling molecules affecting the activity of chromatin-modifying enzymes. However, how metabolism coordinates cell-state changes, especially in human pre-implantation development, remains unclear. Here we uncover that inducing naive human embryonic stem cells towards the trophectoderm lineage results in considerable metabolic rewiring, characterized by aKG accumulation. Elevated aKG levels potentiate the capacity of naive embryonic stem cells to specify towards the trophectoderm lineage. Moreover, increased aKG levels promote blastoid polarization and trophectoderm maturation. aKG supplementation does not affect global histone methylation levels; rather, it decreases acetyl-CoA availability, reduces histone acetyltransferase activity and weakens the pluripotency network. We propose that metabolism functions as a positive feedback loop aiding in trophectoderm fate induction and maturation, highlighting that global metabolic rewiring can promote specificity in cell fate decisions through intricate regulation of signalling and chromatin. Overall design: scRNA-seq of H9 human embryonic stem cells cultured in tt2iLIFGö and hiTSCs. tt2iLIFGö cells were treated for 24hrs with 0 or 4mM dimethyl-alphaketoglutarate. hiTSCs were derived from treated or untreated tt2iLIFGö cells and collected after 72 hours. HTO barcode sequence hashtagged sample TotalSeq-A0253 anti-human Hashtag 3 TTCCGCCTCTCTTTG tt2iLIFGo_DMSO TotalSeq-A0254 anti-human Hashtag 4 AGTAAGTTCAGCGTA tt2iLIFGo_dm-aKG TotalSeq-A0255 anti-human Hashtag 5 AAGTATCGTTTCGCA hiTSC_DMSO TotalSeq-A0256 anti-human Hashtag 6 GGTTGCCAGATGTCA hiTSC_dm-aKG