Huang Xingyun, Yu Jiamin, Gou Shixue, Qin Hongyu, Lu William W, Li Zhen, Tong Liping, Chen Di
Research Center for Computer-aided Drug Discovery, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, Guangdong 518055, China.
Faculty of Pharmaceutical Sciences, Shenzhen University of Advanced Technology, Shenzhen, Guangdong 518055, China.
Genes Dis. 2024 Nov 16;12(4):101468. doi: 10.1016/j.gendis.2024.101468. eCollection 2025 Jul.
Osteoarthritis (OA) is a chronic degenerative joint disease. Currently, OA is incurable. Abnormal activation of canonical Wnt/β-catenin or Indian hedgehog (Ihh) signaling could lead to OA development and progression. This study aimed to determine if targeting β-catenin and Ihh signaling could yield an effective therapeutic intervention for OA disease. CRISPR/CasRx is a new RNA interference tool that can precisely and efficiently cleave single-strand RNAs. In this study, we screened CRISPR-derived RNA (crRNA) targeting and and selected two optimal crRNAs for each gene. CasRx-mediated and knockdown showed high efficiency and specificity with no obvious off-target effects . We then performed intra-articular injection of selected crRNAs driven by the adeno-associated virus into an OA mouse model. Micro-CT, histological, and histomorphometric analyses were conducted to evaluate the efficacy of CasRx approach on OA treatment. We found that the knockdown of and decelerated pathological damage in the keen joint of the experimental OA mouse model. Our findings suggest that CasRx-mediated and knockdown could be a potential strategy for OA treatment.
骨关节炎（OA）是一种慢性退行性关节疾病。目前，OA无法治愈。经典Wnt/β-连环蛋白或印度刺猬（Ihh）信号通路的异常激活可导致OA的发生和发展。本研究旨在确定靶向β-连环蛋白和Ihh信号通路是否能为OA疾病带来有效的治疗干预。CRISPR/CasRx是一种新型RNA干扰工具，可精确、高效地切割单链RNA。在本研究中，我们筛选了靶向[基因名称1]和[基因名称2]的CRISPR衍生RNA（crRNA），并为每个基因选择了两个最佳crRNA。CasRx介导的[基因名称1]和[基因名称2]敲低显示出高效率和特异性，且无明显脱靶效应。然后，我们将腺相关病毒驱动的选定crRNA关节腔内注射到OA小鼠模型中。进行了微型计算机断层扫描（Micro-CT）、组织学和组织形态计量学分析，以评估CasRx方法对OA治疗的疗效。我们发现，[基因名称1]和[基因名称2]的敲低减缓了实验性OA小鼠模型膝关节的病理损伤。我们的研究结果表明，CasRx介导的[基因名称1]和[基因名称2]敲低可能是一种潜在的OA治疗策略。
