miR-3606-3p alleviates skin fibrosis by integratively suppressing the integrin/FAK, p-AKT/p-ERK, and TGF-β signaling cascades

Yahui Chen; Yiyi Gong; Mengkun Shi; Haoxing Zhu; Yulong Tang; Delin Huang; Wei Wang; Chenyi Shi; Xueyi Xia; Ying Zhang; Jianlan Liu; Jia Huang; Mengguo Liu; Huyan Chen; Yanyun Ma; Ziyu Wang; Lei Wang; Wenzhen Tu; Yinhuan Zhao; Jinran Lin; Li Jin; Jörg Hw Distler; Wenyu Wu; Jiucun Wang; Xiangguang Shi

doi:10.1016/j.jare.2024.11.027

miR-3606-3p alleviates skin fibrosis by integratively suppressing the integrin/FAK, p-AKT/p-ERK, and TGF-β signaling cascades

J Adv Res. 2024 Nov 20:S2090-1232(24)00546-0. doi: 10.1016/j.jare.2024.11.027. Online ahead of print.

Authors

Yahui Chen¹, Yiyi Gong¹, Mengkun Shi², Haoxing Zhu¹, Yulong Tang³, Delin Huang³, Wei Wang³, Chenyi Shi³, Xueyi Xia¹, Ying Zhang¹, Jianlan Liu¹, Jia Huang¹, Mengguo Liu¹, Huyan Chen¹, Yanyun Ma³, Ziyu Wang³, Lei Wang⁴, Wenzhen Tu⁴, Yinhuan Zhao⁴, Jinran Lin¹, Li Jin³, Jörg Hw Distler⁵, Wenyu Wu⁶, Jiucun Wang⁷, Xiangguang Shi⁸

Affiliations

¹ Department of Dermatology, Huashan Hospital and Human Phenome Institute, Fudan University, Shanghai, China.
² Department of Thoracic Surgery, Huashan Hospital & Cancer Metastasis Institute, Fudan University, Shanghai, China.
³ State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai, China.
⁴ Division of Rheumatology, Shanghai TCM-Integrated Hospital, Shanghai, China.
⁵ University Hospital Düsseldorf and Heinrich-Heine University, Düsseldorf, Germany.
⁶ Department of Dermatology, Huashan Hospital, Shanghai Institute of Dermatology, State Key Laboratory of Molecular Engineering of Polymers, Fudan University, Shanghai, China; Department of Dermatology, Jing'an District Central Hospital, Shanghai, China; National Clinical Research Center for Aging and Medicine, Huashan Hospital, Fudan University, Shanghai, China; Ministry of Education Key Laboratory of Contemporary Anthropology, School of Life Sciences, and Academy for Engineering and Technology, Fudan University, Shanghai, China. Electronic address: wuwenyu@huashan.org.cn.
⁷ Department of Dermatology, Huashan Hospital and Human Phenome Institute, Fudan University, Shanghai, China; State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai, China; Deptartment of Allergy and Immunology, Huashan Hospital, and Research Center of Allergy and Diseases, Fudan University, Shanghai, China; Research Unit of Dissecting the Population Genetics and Developing New Technologies for Treatment and Prevention of Skin Phenotypes and Dermatological Diseases (2019RU058), Chinese Academy of Medical Sciences, Shanghai, China. Electronic address: jcwang@fudan.edu.cn.
⁸ Department of Dermatology, Huashan Hospital and Human Phenome Institute, Fudan University, Shanghai, China. Electronic address: guangxiangshi@163.com.

PMID: 39571732
DOI: 10.1016/j.jare.2024.11.027

Abstract

Introduction: Fibroblast abnormalities are crucial causes of skin fibrosis, including systemic sclerosis (SSc) and keloids. However, their mechanisms, including underlying microRNA regulatory mechanisms, remain elusive.

Objectives: This study aimed to evaluate the roles, mechanisms, and therapeutic potential of miR-3606-3p in regulating multiple fibroblast abnormalities.

Methods: The miR-3606-3p levels were evaluated in skin tissues and primary fibroblasts. RNA-seq and luciferase assays were employed to identify miR-3606-3p targets. Collagen contraction, western blotting, in vivo imaging, and real-time cellular analysis were used to assess fibroblast abnormalities. The therapeutic potential of miR-3606-3p was evaluated in mice.

Results: MiR-3606-3p decreased in skin tissues (SSc: Fold Change (FC) = - 2.95, P = 0.0101; keloid: FC = - 3.42, P < 0.0001) and primary fibroblasts (SSc: FC = - 12.74, P = 0.0278; keloid: FC = - 2.08, P = 0.0021) from skin fibrosis patients, and negatively correlated with disease severity. Mechanistically, miR-3606-3p targeted the 3'-untranslated regions (3'-UTRs) of Integrin αV (ITGAV), GRB2-associated binding protein 1 (GAB1), and transforming growth factor beta receptor 2 (TGFBR2), all of these three targets increased in skin fibrosis. Simultaneously, miR-3606-3p inhibited fibroblast's fibrogenesis, migration, inflammation, and proliferation by inhibiting ITGAV/integrin/FAK, GAB1/p-AKT/p-ERK, and TGFBR2/p-SMAD2/3 signaling. ITGAV-mediated integrin/FAK signaling unidirectionally activated the p-AKT/p-ERK and p-SMAD2/3 pathways. Knockdown of GAB1 and TGFRB2 reduced ITGAV-induced p-AKT/p-ERK and p-SMAD2/3 activities. MiR-3606-3p, si-ITGAV, si-GAB1, and si-TGFBR2 exhibited significant inhibition of fibrogenesis and migration. Inflammation was primarily inhibited by si-ITGAV and si-GAB1, while proliferation was primarily inhibited by si-TGFBR2. Moreover, miR-3606-3p significantly attenuates skin fibrosis in keloid-bearing mice.

Conclusions: MiR-3606-3p is downregulated in skin fibrosis. Moreover, it negatively correlates with disease severity. Functionally, miR-3606-3p inhibits fibrogenesis, migration, inflammation, and proliferation of fibroblasts. Mechanistically, miR-3606-3p inhibits ITGAV, GAB1, and TGFBR2 by targeting their 3'-UTRs. ITGAV-, GAB1-, and TGFBR2-activated integrin/AKT/ERK/SMAD2/3 signaling induced fibroblast abnormalities. In vivo, miR-3606-3p inhibits skin fibrosis in mice. Therefore, the multi-targeting, multi-phenotypic regulatory properties of miR-3606-3p suggest its potential utility in clinical treatment.

Keywords: Fibroblast abnormalities; GAB1; ITGAV; Skin fibrosis; TGFBR2; miR-3606-3p.