Serine metabolism during differentiation of human iPSC-derived astrocytes

Astrocytes are essential players in development and functions, being particularly relevant as regulators of brain energy metabolism, ionic homeostasis and synaptic transmission. They are also the major source of l-serine in the brain, which is synthesized from the glycolytic intermediate 3-phosphoglycerate through the phosphorylated pathway. l-Serine is the precursor of the two main co-agonists of the N-methyl-d-aspartate receptor, glycine and d-serine. Strikingly, dysfunctions in both l- and d-serine metabolism are associated with neurological and psychiatric disorders. Here, we exploited a differentiation protocol, based on the generation of human mature astrocytes from neural stem cells, and investigated the modification of the proteomic and metabolomic profile during the differentiation process. We show that differentiated astrocytes are more similar to mature rather than to reactive ones, and that axogenesis and pyrimidine metabolism increase up to 30 days along with the folate cycle and sphingolipid metabolism. Consistent with the proliferation and cellular maturation processes that are taking place, also the intracellular levels of l-serine, glycine, threonine, l- and d-aspartate (which level is unexpectedly higher than that of d-serine) show the same biosynthetic time course. A significant utilization of l-serine from the medium is apparent while glycine is first consumed and then released with a peak at 30 days, parallel to its intracellular level. These results underline how metabolism changes during astrocyte differentiation, highlight that d-serine synthesis is restricted in differentiated astrocytes and provide a valuable model for developing potential novel therapeutic approaches to address brain diseases, especially the ones related to serine metabolism alterations.

Automated summary

Astrocytes play important roles in brain development and functions, including regulation of brain energy metabolism, ionic homeostasis, and synaptic transmission. Dysfunctions in metabolism of l-serine and d-serine are associated with neurological and psychiatric disorders. In this study, the differentiation process of human mature astrocytes from neural stem cells was investigated, revealing changes in proteomic and metabolomic profiles. The results show that differentiated astrocytes resemble mature rather than reactive ones, and reveal increased axogenesis and pyrimidine metabolism, as well as changes in the folate cycle and sphingolipid metabolism. These findings provide insights into astrocyte differentiation and offer a valuable model for studying serine metabolism alterations in brain diseases.