Glutamine deprivation enhances metabolic reprogramming during kidney organoid directed differentiation

In this study, we systematically performed comprehensive metabolite and transcriptome profiling of human pluripotent stem cells (hPSCs) during differentiation into NPCs and later into multicellular organoids. We found activation of distinct metabolic pathways during early stage progression from hPSCs to NPCs; however, NPCs, organoids, and the intervening developmental stages between them shared similar metabolic profiles across this later stage of maturation. Importantly, the Alanine-aspartate-glutamate and glutamine pathways were significantly altered during the first stages of differentiation. Moreover, hPSCs survived glutamine deprivation during in vitro differentiation, and NPCs were successfully generated both in the absence and presence of glutamine and glutamate. Surprisingly, metabolic deprivation resulted in enhanced maturation, specifically in podocytes, as evidenced through single cell RNA-Seq analysis. These findings highlight the critical regulatory role of glutamine metabolism in the derivation of nephron progenitor cells and kidney organoids. Overall design: We have used 10x Genomics to perform single cell sequencing on organoid cells with and without glutamine deprivation