TGFβ1 selectively activates pro-fibrotic gene expression in gingival fibroblasts.

YAP inhibitor celastrol blocks TGFβ1-induced activation of gingival fibroblasts.

YAP inhibitors deserve future consideration to impair gingival overgrowth.

To 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.

Human 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.

Celastrol 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.

YAP1 inhibitors such as celastrol could be used to impair pro-fibrotic responses to TGFβ1 in human gingival fibroblasts.

cellular communication network factor 2

cellular communication network factor 1

C-X-C motif chemokine 12

dimethyl sulfoxide, DAPI, 4′,6-diamidino-2-phenylindole, ECM, extracellular matrix

enhanced chemiluminescence

early growth response-1

matrix metalloproteinase

phosphate buffered saline

procollagen-lysine,2-oxoglutarate 5-dioxygenase 2

real time polymerase chain reaction

SRY-related HMG-box genes 9

tris buffered saline with tween 20, TGFβ, transforming growth factor β

yes-associated protein

Fibroblast

Gingiva

Mechanotransduction

Celastrol

YAP1

TGFβ

© 2024 The Author(s). Published by Elsevier Ltd.