2. Academic Validation
  3. Inhibition of enhancer of zeste homolog 2 prevents corneal myofibroblast transformation in vitro

Inhibition of enhancer of zeste homolog 2 prevents corneal myofibroblast transformation in vitro

  • Exp Eye Res. 2021 Jul;208:108611. doi: 10.1016/j.exer.2021.108611.
Kai Liao 1,  Zekai Cui 2,  Yong Zeng 3,  Jian Liu 2,  Yini Wang 2,  Zhijie Wang 1,  Shibo Tang 4,  Jiansu Chen 5
Affiliations

Affiliations

  • 1 Aier School of Ophthalmology, Central South University, Changsha, Hunan, China; Aier Eye Institute, Changsha, Hunan Province, China.
  • 2 Aier Eye Institute, Changsha, Hunan Province, China.
  • 3 Department of Ophthalmology, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, Chengdu, China.
  • 4 Aier School of Ophthalmology, Central South University, Changsha, Hunan, China; Aier Eye Institute, Changsha, Hunan Province, China; CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai, China. Electronic address: [email protected].
  • 5 Aier School of Ophthalmology, Central South University, Changsha, Hunan, China; Aier Eye Institute, Changsha, Hunan Province, China; Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, China. Electronic address: [email protected].
PMID: 33992624 DOI: 10.1016/j.exer.2021.108611
Abstract

Purpose: Corneal fibroblast can be transformed into corneal myofibroblasts by TGF-β1. Enhancer of zeste homolog 2 (EZH2) upregulation has been observed in the occurrence of other fibrotic disorders. We investigated the role of EZH2 in the progression of corneal fibrosis and the antifibrotic effect of EZH2 inhibition in corneal fibroblasts (CFs).

Methods: Primary CFs were isolated from corneal limbi and the CFs were treated with TGF-β1 to induce fibrosis. EPZ-6438 and EZH2 siRNA were used to inhibit EZH2 expression. Myofibroblast activation and extracellular matrix (ECM) protein synthesis was detected by quantitative Real-Time PCR, western blotting, and immunofluorescence staining assay. The functions of myofibroblast were evaluated by cell migration and collagen gel contraction assays. Molecular mechanisms involved in EZH2 inhibition were investigated by RNA sequencing.

Results: TGF-β1 activated EZH2 expression in CFs. Treatment with EPZ-6438 (5 μM) and EZH2 siRNA considerably suppressed corneal myofibroblast activation and ECM protein synthesis in CFs induced by TGF-β1 when compared to the control group. EPZ-6438 (5 μM) suppressed cell migration and gel contraction in CFs. RNA sequencing results revealed that antifibrotic genes were activated after EZH2 inhibition to suppress corneal myofibroblast activation.

Conclusion: Inhibition of EZH2 suppresses corneal myofibroblast activation and ECM protein synthesis, and could serve as a novel therapeutic target for preventing corneal scarring.

Keywords

Corneal fibroblast; EZH2; Fibrosis; Transcriptomics.

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