a-Ketoglutarate promotes trophectoderm induction and maturation from naive human embryonic stem cells [scRNA-Seq 3D]

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 Aggrewells in blastoid induction medium for 40h or 120h. Aggregates or blastoids were treated for 40h with 0 or 4mM dimethyl-alphaketoglutarate.