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Effect of connective tissue growth factor gene editing using adeno-associated virus–mediated CRISPR–Cas9 on rabbit glaucoma filtering surgery outcomes

Gene Therapy volume 28, pages 277–286 (2021)Cite this article

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Abstract

Suppressing excessive wound healing responses is critical to ensure surgical success in glaucoma filtration surgery (GFS). Currently used adjunctive materials can lead to side effects due to the nonselectivity in cell inhibition and may require repeated applications. The clustered regularly interspaced short palindromic repeats (CRISPR)–Cas9 system may become a compelling opportunity in glaucoma surgery due to its high selectivity and permanent effect. Connective tissue growth factor (CTGF) is one of the most potent stimulators of tissue fibrosis in the eye. Therefore, we tested the effect of CTGF suppression using the CRISPR–Cas9 system on GFS fibrosis. We used an adeno-associated virus (AAV)–CRISPR–Cas9 system and confirmed successful CTGF suppression was achieved in fibroblasts in vitro through western blot analysis and deep sequencing. In the in vivo intereye-comparison rabbit GFS model, CRISPR–CTGF-treated eyes showed significantly better survival of the surgery site, less subconjunctival fibrosis, limited collagen deposition, and reduced cellularity than untreated eyes. Our results suggest a new possibility of CRISPR–Cas9-mediated CTGF suppression to improve human GFS outcomes.

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Fig. 1: In vitro suppression of CTGF expression by the CRISPR genome-editing system.
Fig. 2: In vivo delivery of viral vectors and suppression of CTGF using the AAV–CRISPR genome-editing system.
Fig. 3: Bleb survival and characteristics in the CRISPR-CTGF-treated and untreated groups.
Fig. 4: Photographs for the bleb appearance in the CRISPR–CTGF-treated and untreated groups.
Fig. 5: Histopathologic and immunohistochemistry results in CRISPR–CTGF-treated and untreated groups.

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Acknowledgements

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2015R1D1A1A01056574).

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Authors and Affiliations

  1. Department of Ophthalmology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea

    Eun Jung Lee, Jong Chul Han, Do Young Park & Changwon Kee

  2. Department of Pathology and Translational Genomics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea

    Junhun Cho

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Lee, E.J., Han, J.C., Park, D.Y. et al. Effect of connective tissue growth factor gene editing using adeno-associated virus–mediated CRISPR–Cas9 on rabbit glaucoma filtering surgery outcomes. Gene Ther 28, 277–286 (2021). https://doi.org/10.1038/s41434-020-0166-4

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