Department of Orthopedic Surgery, Yonsei University College of Medicine, Seoul 03722, South Korea.
Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, South Korea.
Int J Med Sci. 2021 Jul 25;18(15):3353-3360. doi: 10.7150/ijms.56760. eCollection 2021.
Despite several studies on the effect of adeno-associated virus (AAV)-based therapeutics on osteoarthritis (OA), information on the transduction efficiency and applicable profiles of different AAV serotypes to chondrocytes in hard cartilage tissue is still limited. Moreover, the recent discovery of additional AAV serotypes makes it necessary to screen for more suitable AAV serotypes for specific tissues. Here, we compared the transduction efficiencies of 14 conventional AAV serotypes in human chondrocytes, mouse OA models, and human cartilage explants obtained from OA patients. To compare the transduction efficiency of individual AAV serotypes, green fluorescent protein (GFP) expression was detected by fluorescence microscopy or western blotting. Likewise, to compare the transduction efficiencies of individual AAV serotypes in cartilage tissues, GFP expression was determined using fluorescence microscopy or immunohistochemistry, and GFP-positive cells were counted. Only AAV2, 5, 6, and 6.2 exhibited substantial transduction efficiencies in both normal and OA chondrocytes. All AAV serotypes except AAV6 and rh43 could effectively transduce human bone marrow mesenchymal stem cells. In human and mouse OA cartilage tissues, AAV2, AAV5, AAV6.2, AAV8, and AAV rh39 showed excellent tissue specificity based on transduction efficiency. These results indicate the differences in transduction efficiencies of AAV serotypes between cellular and tissue models. Our findings indicate that AAV2 and AAV6.2 may be the best choices for AAV-mediated gene delivery into intra-articular cartilage tissue. These AAV vectors hold the potential to be of use in clinical applications to prevent OA progression if appropriate therapeutic genes are inserted into the vector.
尽管已有多项研究探讨了腺相关病毒（AAV）基疗法对骨关节炎（OA）的影响，但对于不同 AAV 血清型在硬软骨组织中的软骨细胞中的转导效率和适用谱，信息仍然有限。此外，最近发现了更多的 AAV 血清型，这使得有必要筛选出更适合特定组织的 AAV 血清型。在这里，我们比较了 14 种常规 AAV 血清型在人软骨细胞、小鼠 OA 模型和从 OA 患者获得的人软骨外植体中的转导效率。为了比较各个 AAV 血清型的转导效率，通过荧光显微镜或 Western blot 检测绿色荧光蛋白（GFP）的表达。同样，为了比较各个 AAV 血清型在软骨组织中的转导效率，通过荧光显微镜或免疫组织化学检测 GFP 的表达，并计数 GFP 阳性细胞。只有 AAV2、5、6 和 6.2 在正常和 OA 软骨细胞中均表现出较高的转导效率。除 AAV6 和 rh43 外，所有 AAV 血清型均可有效转导人骨髓间充质干细胞。在人和小鼠 OA 软骨组织中，根据转导效率，AAV2、AAV5、AAV6.2、AAV8 和 AAV rh39 显示出良好的组织特异性。这些结果表明 AAV 血清型在细胞和组织模型之间的转导效率存在差异。我们的研究结果表明，AAV2 和 AAV6.2 可能是 AAV 介导的基因递送至关节内软骨组织的最佳选择。如果将适当的治疗基因插入载体，这些 AAV 载体有可能在预防 OA 进展的临床应用中发挥作用。
