Mechanisms of carbon dot-guided rAAV gene vector formulation and release forimproved, safe rAAV-mediated therapeutic gene transfer in human cartilage lesions.

局部软骨创伤性病变是骨科中的关键问题，因为成人关节软骨由于该组织中缺乏血管化而不能完全自行愈合。鉴于软骨完全自愈的能力有限，已经开发了各种选项来改善软骨修复，但没有一种可以完全治愈软骨损伤。在局灶性软骨缺损中施用有效的、临床适应的重组腺相关病毒（rAAV）载体是在损伤部位暂时和空间转移软骨修复基因以延长基因产物表达的强有力方法。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软骨修复蛋白的治疗性rAAV载体能够抑制软骨修复蛋白表达的假设。（增殖，促合成代谢）基因（软骨特异性性别决定区Y型高迁移率盒9 -SOX 9-转录因子和转化生长因子β- TGF-β）可以通过碳量子点（CD）作为保护性的受控递送系统来递送，相对于直接应用游离形式的载体，在体外天然3D环境中，在人实验性骨软骨缺损的原位翻译前模型中，以及在体内临床相关的翻译局灶性软骨缺损中，有效且持久地增强原代人骨髓来源的间充质基质细胞中软骨修复相关的过程和机制。该项目可能在不久的将来提供新的，有效的治疗方法来增强患者的软骨修复。

Focal cartilage traumatic lesions are critical issues in orthopaedics since the adult articular cartilage does not completely heal on itself as a result of the lack of vascularization in this tissue. In light of the restricted ability of the cartilage for full self-healing, various options have been developed to improve cartilage repair, but none can fully heal cartilage lesions. Administration of the potent, clinically adapted recombinant adeno-associated virus (rAAV) vectors in focal cartilage defects is a strong approach to temporarily and spatially transfer chondroreparative 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 chondroreparative (proliferative, pro-anabolic) genes (the cartilage-specific sex-determining region Y-type high mobility box 9 - SOX9 - transcription factor and the transforming growth factor beta - TGF-ß) may be delivered via carbon dots (CDs) as protective, controlled delivery systems to safely, effectively, and durably enhance the processes and mechanisms relevant of cartilage repair in primary human bone marrow-derived mesenchymal stromal cells in a natural 3D environment in vitro, in a pre-translational model of human experimental osteochondral defect in situ, and in clinically relevant, translational focal cartilage defects in vivo relative to direct application of the vectors in their free form. This project may offer new, effective therapies to enhance cartilage repair in patients in a close future.