Gfpt2 Regulates Fibroblast Activation by Glutathione Metabolism

ABSTRACT: Aims: During fibroblast activation, the cell undergoes metabolic reprogramming to increase glycolysis and support increased cellular growth. The role of many ancillary metabolic pathways in cardiac fibroblasts remains poorly defined. Here we investigate the role of Gfpt2, the rate-limiting enzyme of the hexosamine biosynthesis pathway, and metabolism in fibroblast activation first in cardiac fibroblasts followed by an exploration in fibroblasts originating from other tissues. Methods and Results: Gfpt2 knockdown in adult murine cardiac fibroblasts resulted in upregulation of myofibroblast related genes and increased contractility indicating a shift towards an activated fibroblast phenotype. Bulk RNA sequencing supported this observation by showing enrichment of extracellular matrix related pathways and gene expression patterns resembling TGFβ stimulated fibroblast activation. Untargeted metabolomics further identified glutathione metabolism as a downstream pathway of Gfpt2. Together with Seahorse Mito Stress Test analysis, these data revealed that Gfpt2 knockdown reduced glutathione and increased reactive oxygen species (ROS), suggesting that Gfpt2 and glutathione regulate fibroblast activation through ROS scavenging. Consistent with this, glutathione treatment following Gfpt2 knockdown or TGFβ stimulation prevented cardiac fibroblast activation. Finally, screening fibroblasts from other tissues demonstrated that the Gfpt2-glutathione regulatory axis is shared across some, but not all, tested fibroblast populations. Conclusions: We identified Gfpt2 as a metabolic regulator of fibroblast activation through glutathione metabolism. This regulatory axis in fibroblasts is shared across a subset of tissues and poses as a promising potential target for the development of feature therapeutics against fibrosis.

ORGANISM(S): Mus musculus

PROVIDER: GSE337431 | GEO | 2026/09/10

REPOSITORIES: GEO