WDR36 regulates trophectoderm differentiation during human preimplantation embryonic development through glycolytic metabolism

Mammalian pre-implantation development is a complex process involving sophisticated regulatory dynamics. WD repeat domain 36 (WDR36) is known to play a critical role in mouse early embryonic development, but its regulatory functions in human embryogenesis is still elusive due to limited access to human embryos. The human pluripotent stem cell-derived blastocyst-like structure, termed a blastoid, offer an alternative means to study human development in a dish. In this study, after verifying that WDR36 inhibition disrupted polarization in mouse early embryos, we further demonstrated that WDR36 interference could block human blastoid formation, dominantly hindering the trophectoderm lineage commitment. Both transcriptomics and targeted metabolomics analyses revealed that WDR36 interference downregulated glucose metabolism. WDR36 can interact with glycolytic metabolic protein LDHA, thereby positively regulating glycolysis during the late stage of human blastoid development. Taken together, our study has established a mechanistic connection between WDR36, glucose metabolism, and cell fate determination during early embryonic lineage commitment, which may provide potential insights into novel therapeutic targets for early adverse pregnancy interventions. We treated iWDR36 iPS cell-derived blastoids with Dox to knockdown WDR36 gene, starting on Day 3. Then, we collected samples at Day 4 and Day 6. We compared the gene expression profiles between these samples as well as transcriptome data set of natural blastocysts from the literature.