Mutual Amplification of GLI2/Hedgehog and Transcription Factor JUN/AP‐1 Signaling in Fibroblasts in Systemic Sclerosis: Potential Implications for Combined Therapies.

Bergmann, Christina; Chenguiti Fakhouri, Sara; Trinh‐Minh, Thuong; Filla, Tim; Rius Rigau, Aleix; Ekici, Arif B.; Merlevede, Benita; Hallenberger, Ludwig; Zhu, Honglin; Dees, Clara; Matei, Alexandru‐Emil; Auth, Janina; Györfi, Andrea‐Hermina; Zhou, Xiang; Rauber, Simon; Bozec, Aline; Dickel, Nicholas; Liang, Chunguang; Kunz, Meik; Schett, Georg

Objective: Deregulation of the cJUN/AP‐1 and hedgehog/GLI2 signaling pathways has been implicated in fibroblast activation in systemic sclerosis (SSc). However, the consequences of their concomitant up‐regulation are unknown. Here, we tested the hypothesis that mutual amplification of both pathways might drive persistent fibroblast activation. Methods: Cultured fibroblasts and skin sections of patients with diffuse SSc and healthy volunteers were analyzed. cJUN/AP‐1 signaling and hedgehog/GLI2 signaling were inhibited using knockdown and pharmacologic approaches. Hedgehog signaling was activated in mice by fibroblast‐specific overexpression of constitutively active Smoothened. Results: cJUN and GLI2 are concomitantly up‐regulated and colocalize in fibroblasts of patients with SSc compared to healthy controls. Activation of hedgehog/GLI2 signaling induces the expression of cJUN in vitro and in vivo, whereas inactivation of GLI2 inhibits cJUN expression. Likewise, inactivation of cJUN impairs the expression of GLI2. This mutual regulation occurs at the level of transcription with binding of cJUN and GLI2 to specific binding motifs. Interference with this mutual amplification of cJUN signaling and GLI2 signaling inhibits fibroblast activation and collagen release: Inhibition of cJUN/AP‐1 signaling ameliorates hedgehog‐induced fibroblast activation and skin fibrosis in SmoACT mice with a reduction of skin thickness of 103% (P = 0.0043) in the treatment group compared to the fibrotic control group. Moreover, combined pharmacologic inhibition of cJUN/AP‐1 and hedgehog/GLI2 exerts additive antifibrotic effects in a model of TGFβ‐driven experimental fibrosis (TBRACT mice). Conclusion: The transcription factors cJUN and GLI2 reinforce each other's activity to promote fibroblast activation in SSc. Interruption of this crosstalk by combined inhibition of both pathways exerts additive antifibrotic effects at well‐tolerated doses.

PROTEIN metabolism; NUCLEAR proteins; PHENOMENOLOGICAL biology; RESEARCH funding; CELLULAR signal transduction; BIOCHEMISTRY; FIBROBLASTS; MICE; SKIN; CELL culture; SYSTEMIC scleroderma; COMBINED modality therapy; ANIMAL experimentation; CASE-control method; COMPARATIVE studies; DNA-binding proteins; CELL receptors

Arthritis & Rheumatology, 2025, Vol 77, Issue 1, p92

2326-5191

Academic Journal

10.1002/art.42979