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Sildenafil increases AAV9 transduction after a systemic administration and enhances AAV9-dystrophin therapeutic effect in mdx mice

Gene Therapy volume 31, pages 19–30 (2024)Cite this article

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Abstract

Adeno-associated virus (AAV) vectors have been successfully used to deliver genes for treating rare diseases. However, the systemic administration of high AAV vector doses triggers several adverse effects, including immune response, the asymptomatic elevation of liver transaminase levels, and complement activation. Thus, improving AAV transduction and reducing AAV dosage for treatment is necessary. Recently, we found that a phosphodiesterase-5 inhibitor significantly promoted AAV9 transduction in vitro by regulating the caveolae and macropinocytosis pathways. When AAV9-Gaussian luciferase (AAV9-Gluc) and AAV9-green fluorescent protein (AAV9-GFP) were injected intravenously into mice pre-treated with sildenafil, the expressions of Gluc in the plasma and GFP in muscle tissues significantly increased (P < 0.05). Sildenafil also improved Evans blue permeation in tissues. Additionally, we found that sildenafil promoted Treg proliferation, inhibited B-cell activation, and decreased anti-AAV9 IgG levels (P < 0.05). Furthermore, sildenafil significantly promoted Duchenne muscular dystrophy gene therapy efficacy using AAV9 in mdx mice; it increased micro-dystrophin gene expression, forelimb grip strength, and time spent on the rotarod test, decreased serum creatine kinase levels, and ameliorated histopathology by improving muscle cell morphology and reducing fibrosis (P < 0.05). These results show that sildenafil significantly improved AAV transduction, suppressed the levels of anti-AAV9 IgG, and enhanced the efficacy of gene therapy.

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Fig. 1: Sildenafil increased the epithelial cell permeability of AAV9 in vitro.
Fig. 2: Sildenafil increased exogenous gene expression mediated by AAV9 vectors in naïve mice.
Fig. 3: Characterization of the prophylactic effect of sildenafil administration on innate immune cells and inflammatory cytokines activated by AAV9 vector delivery in mice.
Fig. 4: Therapeutic effect of sildenafil combined with AAV9-dystrophin vector in mdx mice.
Fig. 5: Sildenafil treatment (50 mg/kg) increased dystrophin transgene expression in the muscles of mdx mice.

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Data availability

The data that support the findings of this study are available from the corresponding authors, XW or XX, upon reasonable request.

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Acknowledgements

We would like to thank Editage (www.editage.cn) for English language editing.

Funding

This work was supported by the Project of Science and Technology Department of Sichuan Province, China (grant number: 18SYXHZ0024), the National Natural Science Foundation of China (grant number: 31901052, 81970171, 82070139), and the Shanghai Committee of Science and Technology, China (grant number: 20ZR1415000).

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Authors and Affiliations

  1. School of Pharmacy, East China University of Science and Technology, Shanghai, China

    Kaiyi Zhou, Meng Yuan, Jiabao Sun, Xiaoying Zong, Zhanao Li, Xiao Xiao & Xia Wu

  2. State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai, China

    Feixu Zhang & Dingyue Tang

  3. The General Hospital of Western Theater Command PLA, Sichuan Province, China

    Lichen Zhou

  4. Belief BioMed, Xuhui District, Shanghai, China

    Jing Zheng

  5. Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC, 27517, USA

    Xiao Xiao

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Contributions

Conceptualization, XW and XX, Methodology, KZ, XW and XX; Investigation, KZ, MY, JS; Writing-Original Draft, KZY; Writing-review & editing, KZ, MY and XW; Funding acquisition, XW, JZ, and LZ.

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Correspondence to Xiao Xiao or Xia Wu.

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All experimental procedures involving animals were approved by the Ethical Committee of the Shanghai Model Organisms Center, Inc. (IACUC:2022-0001-06).

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Zhou, K., Yuan, M., Sun, J. et al. Sildenafil increases AAV9 transduction after a systemic administration and enhances AAV9-dystrophin therapeutic effect in mdx mice. Gene Ther 31, 19–30 (2024). https://doi.org/10.1038/s41434-023-00411-3

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