The yes-associated protein-1 (YAP1) inhibitor celastrol suppresses the ability of transforming growth factor β to activate human gingival fibroblasts

Abstract

ObjectiveTo determine whether celastrol, an inhibitor of the mechanosensitive transcriptional cofactor yes-associated protein-1 (YAP1), impairs the ability of TGFβ1 to stimulate fibrogenic activity in human gingival fibroblast cell line. DesignHuman gingival fibroblasts were pre-treated with celastrol or DMSO followed by stimulation with or without TGFβ1 (4 ng/ml). We then utilized bulk RNA sequencing (RNAseq), real-time polymerase chain reaction (RT-PCR), Western blot, immunofluorescence, cell proliferation assays to determine if celastrol impaired TGFβ1-induced responses in a human gingival fibroblast cell line. ResultsCelastrol impaired the ability of TGFβ1 to induce expression of the profibrotic marker and mediator CCN2. Bulk RNAseq analysis of gingival fibroblasts treated with TGFβ1, in the presence or absence of celastrol, revealed that celastrol impaired the ability of TGFβ1 to induce mRNA expression of genes within extracellular matrix, wound healing, focal adhesion and cytokine/Wnt signaling clusters. RT-PCR analysis of extracted RNAs confirmed that celastrol antagonized the ability of TGFβ1 to induce expression of genes anticipated to contribute to fibrotic responses. Celastrol also reduced gingival fibroblast proliferation, and YAP1 nuclear localization in response to TGFβ1. ConclusionYAP1 inhibitors such as celastrol could be used to impair pro-fibrotic responses to TGFβ1 in human gingival fibroblasts.