Mechanisms of alginate/rAAV gene vector encapsulation and release for enhanced, saferAAV-mediated therapeutic gene transfer in human meniscal lesions

半月板是膝关节中的重要组织，半月板的病变是运动医学中高度普遍的未解决的问题，如果不治疗，可能使邻近的关节软骨易患骨关节炎。鉴于半月板完全自我愈合的能力有限，特别是在其无血管区，已经开发了各种选择来改善半月板修复，但没有一种可以完全愈合半月板损伤，这也受到临床中同种异体移植物/替代物的可用性、排斥和/或失败的限制。施用有效的、临床上适应的重组腺相关病毒（rAAV）载体于直肠病变中是在损伤部位暂时和空间转移直肠修复基因以延长基因产物表达的强有力的方法。然而，rAAV的安全、临床使用仍然受到体内各种障碍的阻碍，特别是受到人类群体中天然存在的针对AAV衣壳蛋白的中和抗体的阻碍。为了解决这个问题，本发明的目的是检验编码神经修复性疾病的治疗性rAAV载体能够抑制肿瘤生长的假设。（增殖，促合成代谢）基因（碱性成纤维细胞生长因子- FGF-2，转化生长因子β- TGF-β）可以通过基于临床批准的藻酸盐的水凝胶作为保护性的受控递送系统来递送，并且相对于直接应用游离形式的载体，在体外原代人椎间盘纤维软骨细胞中、在人实验性椎间盘损伤原位的翻译前模型中以及在临床相关的体内翻译性椎间盘损伤中持久地增强椎间盘修复相关的过程和机制。该项目可能会提供新的，有效的治疗方法，以加强患者在不久的将来修复。

Lesions to the meniscus, a vital tissue in the knee joint, are highly prevalent, unsolved problems in sports medicine and may predispose the adjacent articular cartilage to osteoarthritis if left untreated. In light of the restricted ability of the meniscus for complete self-healing especially in its avascular zone, various options have been developed to improve meniscal repair, but none can fully heal meniscal lesions, being also limited by the availability, rejection, and/or failure of allograft/substitutes in the clinics. Administration of the potent, clinically adapted recombinant adeno-associated virus (rAAV) vectors in meniscal lesions is a strong approach to temporarily and spatially transfer meniscal reparative genes in sites of injury for an extended expression of the gene products. Yet, the safe, clinical use of rAAV is still hindered by various obstacles in vivo, especially by the natural presence of neutralizing antibodies against the AAV capsid proteins in the human population. To address this issue, the goal of the present proposal is to test the hypothesis that therapeutic rAAV vectors coding for meniscal reparative (proliferative, pro-anabolic) genes (basic fibroblast growth factor - FGF-2, transforming growth factor beta - TGF- ß) may be delivered via hydrogels based on clinically approved alginate as protective, controlled delivery systems to safely, effectively, and durably enhance the processes and mechanisms relevant of meniscal repair in primary human meniscal fibrochondrocytes in vitro, in a pre-translational model of human experimental meniscal lesion in situ, and in clinically relevant, translational meniscal lesions in vivo relative to direct application of the vectors in their free form. This project may offer new, effective therapies to enhance meniscal repair in patients in a close future.