Enhancing Collagen Biosynthesis in Mammalian Cells Through Hypoxia-Mimetic Prolyl Hydroxylase Inhibition

Collagen, the most abundant protein in the extracellular matrix of mammalian cells, is extensively needed in various biotechnological and therapeutic applications, such as tissue engineering and regeneration, cosmetics, and cultivated meat. Despite the increasing demand for natural collagen from non-animal sources, it is mainly produced from animal connective tissues. Recent research has highlighted that under hypoxia, the activation of the hypoxia-inducible factor (HIF) leads to enhanced collagen type I biosynthesis. However, under normal oxygen conditions, HIF activity is downregulated by the HIF-prolyl hydroxylase (PHD) enzyme. We, therefore, hypothesized that inhibiting PHD could elevate HIF transcriptional activity and enhance collagen biosynthesis under normoxia. Our study demonstrates that inhibiting PHD using exogenous small molecules boosts HIF activity and upregulates the key enzymes, collagen prolyl 4-hydroxylases and lysyl hydroxylases, resulting in up to 29-fold increase in collagen type I in embryonic mouse fibroblast NIH/3T3 cells. These findings suggest that targeting PHD can effectively enhance collagen production in mammalian cells. Therefore, modulating key protein signaling pathways presents a promising strategy for enhancing the production of high-yield natural collagen.