Inhibition of Posterior Capsule Opacification by Adenovirus-Mediated Delivery of Short Hairpin RNAs Targeting TERT in a Rabbit Model

Abstract

Purpose

Posterior capsule opacification (PCO) is the most common postoperative complication after cataract surgery and cannot yet be eliminated. Here, we investigated the inhibitory effects of telomerase reverse transcriptase (TERT) gene silencing on PCO in a rabbit model.

Methods

After rabbit lens epithelial cells (LECs) were treated with adenovirus containing short hairpin RNAs (shRNA) targeting TERT (shTERT group), adenovirus containing scramble nonsense control shRNA (shNC group) or PBS (control group), quantitative real-time polymerase chain reaction and Western blotting were used to measure the expression levels of TERT, and a scratch assay was performed to assess the LEC migration. New Zealand white rabbits underwent sham cataract surgery followed by an injection of adenovirus carrying shTERT into their capsule bag. The intraocular pressure and anterior segment inflammation were evaluated on certain days, and EMT markers (α-SMA and E-cadherin) were evaluated by Western blotting and immunofluorescence. The telomerase activity of the capsule bag was detected by ELISA. At 28 d postoperatively, hematoxylin and eosin staining of the cornea and iris and electron microscopy of the posterior capsule were performed.

Results

Application of shTERT to LECs downregulated the expression levels of TERT mRNA and protein. The scratch assay results showed a decrease in the migration of LECs in the shTERT group. In vivo, shTERT decreased PCO formation after cataract surgery in rabbits and downregulated the expression of EMT markers, as determined by Western blotting and immunofluorescence. In addition, telomerase activity was suppressed in the capsule bag. Despite slight inflammation in the iris, histologic results revealed no toxic effects in the cornea and iris.

Conclusion

TERT silencing effectively reduces the migration and proliferation of LECs and the formation of PCO. Our findings suggest that TERT silencing may be a potential preventive strategy for PCO.

Keywords:

• Cataract surgery
• posterior capsule opacification
• lens epithelial cells
• TERT
• gene therapy

Author contributions

Na He designed the experiments and wrote the manuscript. Na He, Xiangxiang Zhang, Peiling Xie, and Jialing He performed experiments and conducted data acquisition and analysis. Zhigang Lv provided the idea for this study and revised the manuscript.

Ethics statement

No human participants were included in this study. All animal procedures were performed following the ARVO statement for the Use of Animals in Ophthalmic and Vision Research, and were approved by the Jinhua Food and Drug Inspection and Testing Institute (AL-JSYJ202117).

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article.

Additional information

Funding

This work was supported by the Public Welfare Technology Research Project of Zhejiang Province under [grant No. LGD20H120001], the Key Project of Social Development of Jinhua Science and Technology Research Program under [grant No. 2018-3-019] and the Major Project of Social Development of Jinhua Science and Technology Research Program under [grant No. 2022-3-059].