Regulator of Cell Cycle Protein (RGCC/RGC-32) Protects Against Pulmonary Fibrosis

Am J Respir Cell Mol Biol. 2021 Oct 20. doi: 10.1165/rcmb.2021-0022OC. Online ahead of print.

Abstract

Some previous studies in tissue fibrosis have suggested a profibrotic contribution from elevated expression of a protein termed either Regulator of Cell Cycle (RGCC) or Response Gene to Complement 32 Protein (RGC-32). Our analysis of public gene expression datasets, by contrast, revealed a consistent decrease in RGCC mRNA levels in association with pulmonary fibrosis. Consistent with this observation, we found that stimulating primary adult human lung fibroblasts with TGF-β in cell culture elevated collagen expression and simultaneously attenuated RGCC mRNA and protein levels. Moreover, overexpression of RGCC in cultured lung fibroblasts attenuated the stimulating effect of TGF-β on collagen levels. Similar to humans with pulmonary fibrosis, the levels of RGCC were also decreased in vivo in lung tissues of wild-type mice challenged with bleomycin in both acute and chronic models. Mice with constitutive RGCC gene deletion accumulated more collagen in their lungs in response to chronic bleomycin challenge than did wild-type mice. RNA-Seq analyses of lung fibroblasts revealed that RGCC overexpression alone had a modest transcriptomic effect, but in combination with TGF-β stimulation, induced notable transcriptomic changes that negated the effects of TGF-β, including on extracellular matrix-related genes. At the level of intracellular signaling, RGCC overexpression delayed early TGF-β-induced Smad2/3 phosphorylation, elevated the expression of total and phosphorylated antifibrotic mediator STAT1, and attenuated the expression of a profibrotic mediator STAT3. We conclude that RGCC plays a protective role in pulmonary fibrosis and that its decline permits collagen accumulation. Restoration of RGCC expression may have therapeutic potential in pulmonary fibrosis.

Keywords: fibroblast; interstitial lung disease; pulmonary fibrosis; regulator of cell cycle protein; response gene to complement 32 protein.