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Co-delivery of IL-1Ra and SOX9 via AAV inhibits inflammation and promotes cartilage repair in surgically induced osteoarthritis animal models

Gene Therapy volume 32, pages 211–222 (2025)Cite this article

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Abstract

Osteoarthritis (OA), a prevalent joint disorder, can lead to disability, with no effective treatment available. Interleukin-1 (IL-1) plays a crucial role in the progression of OA, and its receptor antagonist (IL-1Ra), a natural IL-1 inhibitor, represents a promising therapeutic target by obstructing the IL-1 signaling pathway. This study delivered IL-1Ra via adeno-associated virus (AAV), a gene therapy vector enabling long-term protein expression, to treat knee osteoarthritis (KOA) in animal models. scAAV-oIL-1Ra-I1/2 injected directly into the joint in both MMT/ACLT-induced KOA model rat improved abnormal gait (increasing footprint area and pressure), subchondral bone lesions, and significantly reduced cartilage wear and pathological scores. In the MMT-induced KOA rabbit model, weight-bearing asymmetry (indicating pain) improved after 8 weeks of scAAV-oIL-1Ra-I1/2 administration, and X-ray showed decreased K-L scores (severity grade), reduced cartilage loss, and lower pathology scores compared to untreated animals. Additionally, sex-determining region Y-type high mobility group box 9 (SOX9) was co-delivered with IL-1Ra via AAV in ACLT + MMT-induced KOA rats. The combined treatment significantly alleviated subchondral bone lesions, cartilage destruction, synovial inflammation, and pathological scores, demonstrating superior efficacy compared to either treatment administered alone. Co-delivering IL-1Ra and SOX9 inhibited IL-1 mediated inflammatory signaling, maintained cartilage homeostasis, and promoted its repair in KOA models, suggesting potential for clinical use.

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Fig. 1: AAVJ4 was screened as the novel joint-tropic AAV capsid for KOA gene therapy. Immunofluorescence staining on mouse joints and β-gal staining on rat DRGs were performed after 3 weeks of intra-articular injection with various rAAV capsids delivering the GFP or LacZ gene.
Fig. 2: AAVJ4 was screened as the novel joint-tropic AAV capsid for KOA gene therapy. Immunofluorescence staining on macaque joints after 4 weeks of intra-articular injection with various rAAV capsids delivering the GFP gene.
Fig. 3: AAVJ4 delivering IL-1Ra effectively inhibited inflammation in vitro.
Fig. 4: scAAV-oIL-1Ra-I1/2 gene therapy improved behavioral performance and mitigated KOA severity in surgery-induced model rats.
Fig. 5: scAAV-oIL-1Ra-I1/2 gene therapy alleviated pain and pathology of joints in MMT-induced model rabbits.
Fig. 6: ssAAV-SOX9 promoted the production of chondrocyte extracellular matrix in vitro.
Fig. 7: Combination gene therapy of scAAV-oIL-1Ra-I1/2 and ssAAV-SOX9 improved KOA pathology in ACLT + MMT-induced model rats.
Fig. 8: Combination gene therapy of scAAV-oIL-1Ra-I1/2 and ssAAV-SOX9 exhibited anti-inflammation and promoted chondrogenesis in KOA rats.

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Data availability

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

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Funding

This work was supported by the National Key Research and Development Program of China (No. 2021YFC2700803).

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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, 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, Xinting Li & Xiao Xiao

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Contributions

Conceptualization, WX and XX, Methodology, ZKY, WX, and XX; Investigation, ZKY, YM, and SJB; Writing—Original Draft, ZKY; Writing—Review & Editing, ZKY, LXT, and WX; Funding Acquisition, WX, and XX.

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Correspondence to Xiao Xiao or Xia Wu.

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We declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Ethical approval and consent to participate

C57BL/6 J mice were obtained from Gempharmatech Co., Ltd. (Nanjing, China). Sprague-Dawley (SD) rats were purchased from Weitong Lihua Co., Ltd. (Beijing, China). New Zealand rabbits were purchased from Aoshima Kangda Aibo Biotechnology Co., Ltd. All experimental procedures involving mouse and rat animals were approved by the Ethical Committee of Shanghai Model Organisms Center, Inc. All experimental procedures involving rabbit animals were approved by the Ethical Committee of PharmaLegacy Laboratories (Shanghai) Co., Ltd. And all experimental procedures involving non-human primates were approved by the Ethical Committee of Kunming Medical University. No. SCXK (Yunnan) K2020-0006, Animal Ethics Code: kmmu20221596. All experiments conformed to relevant regulatory standards.

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Zhou, K., Yuan, M., Sun, J. et al. Co-delivery of IL-1Ra and SOX9 via AAV inhibits inflammation and promotes cartilage repair in surgically induced osteoarthritis animal models. Gene Ther 32, 211–222 (2025). https://doi.org/10.1038/s41434-025-00515-y

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