CRISPR/Cas9基因编辑PLGA/Lipid纳米可视递送系统靶向治疗骨关节炎的作用机制研究

骨关节炎是临床治疗的难题，发现骨关节炎治疗的关键靶点、将阻断该靶点的效应分子高效递送到治疗部位是该领域研究的关键科学问题。课题组前期研究证实YAP siRNA可有效抑制软骨细胞退变并逆转小鼠骨关节炎的进程，提示阻断Hippo/Yap通路可能是骨关节炎治疗的新途径;为解决效应分子递送问题，课题组研制出粒径在300nm左右的PLGA/Lipid复合纳泡，并实现了纳泡载药控释、自由被细胞摄取、干细胞示踪及穿透血脑屏障等功能。本项目拟进一步制备PEI修饰的阳离子PLGA/Lipid纳泡，通过脱水缩合反应共价连接胶原靶向多肽COLBP，利用静电吸附CRISPR/Cas9基因编辑敲除YAP的质粒，构建软骨靶向YAP基因编辑PLGA/Lipid纳米可视递送系统,利用超声评估递送系统的靶向性能并促进质粒递送与跨膜转运，探究超声联合YAP基因编辑对骨关节炎的治疗作用及机制,为骨关节炎治疗提供新策略。

Osteoarthritis is one of the challenges in orthopedic clinical field. The key of the osteoarthritis treatment is to find the target genes of osteoarthritis and efficiently deliver the effect molecules that can effectively block the target gene to the therapeutic site. Previous studies of our group found that YAP siRNA could effectively inhibit the degeneration of chondrocytes and reversed the progress of osteoarthritis in mice, suggesting that YAP is one of the key targets for the treatment of osteoarthritis. Blocking the Hippo/Yap signaling pathway may be a new therapeutic approach for the treatment of osteoarthritis. Meanwhile, our group developed a new PLGA/Lipid composite nanobubble with a particle size of about 300 nm, which showed the functions of drug delivery, controlled release, penetrating blood-brain barrier, and free uptake by cells. This project intends to design a cartilage-targeted CRISPR/cas9 gene editing visual PLGA/lipid nanobubble delivery system. The collagen targeting peptide COLBP is covalently connected onto the PEI-modified PLGA/lipid complex nano-bubbles, and electrostatically adsorbs plasmids carrying the molecular knocking out YAP by the CRISPR/Cas9 gene editing technology. Ultrasound is used to visually monitor the targeting property of the nano-bubbles and promote plasmids delivery and trans-membrane transport. The therapeutic effect and mechanism of ultrasound combined with YAP gene editing on osteoarthritis are studied, providing a new strategy for the treatment of osteoarthritis.