PEO-PPO-PEO Carriers for rAAV-Mediated Transduction of Human Articular Chondrocytes in Vitro and in a Human Osteochondral Defect Model.
PEO-PPO-PEO Carriers for rAAV-Mediated Transduction of Human Articular Chondrocytes in Vitro and in a Human Osteochondral Defect Model.
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DOI：
10.1021/acsami.6b06509
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发表时间：
2016-08
影响因子：
9.5
通讯作者：
A. Rey-Rico;Janina Frisch;J. K. Venkatesan;G. Schmitt;Isabel Rial-Hermida;P. Taboada;A. Concheiro;H. Madry;C. Alvarez‐Lorenzo;M. Cucchiarini
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文献类型：
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作者：
A. Rey-Rico;Janina Frisch;J. K. Venkatesan;G. Schmitt;Isabel Rial-Hermida;P. Taboada;A. Concheiro;H. Madry;C. Alvarez‐Lorenzo;M. Cucchiarini
基因治疗是持久治疗人类骨关节炎（OA）的一种有吸引力的策略，骨关节炎是一种渐进的、不可逆转的关节疾病。基于小型人腺相关病毒（AAV）的基因载体在长时间内原位修饰受损的人关节软骨中表现出主要功效。然而，由于大多数患者存在针对病毒衣壳元件的中和抗体，重组腺相关病毒（rAV）载体的临床应用仍然很复杂。本研究的目的是评估通过聚合物胶束将rAAV载体递送到体外人OA软骨细胞和骨软骨缺损实验模型中以保护基因转移免受实验中和的可行性。采用等温滴定量热法研究了重组腺相关病毒（rAAV）与线性（泊洛沙姆PF 68）或X型（泊洛沙姆T908）聚环氧乙烷（PEO）和聚环氧丙烷（PPO）共聚物胶束（PEO-PPO-PEO胶束）的相互作用。胶束封装允许增加rAAV载体的稳定性和生物活性，并促进更高水平的安全转基因（lacZ）表达在体外和实验性骨软骨缺损相比，无有害影响的细胞或其表型的生物活性的自由载体治疗。值得注意的是，当在所有评价的系统中经由PEO-PPO-PEO胶束递送rAAV时，特别是当使用T908时，也提供了针对抗体中和的保护。总之，这些发现表明PEO-PPO-PEO胶束作为有效工具的潜力，以改善目前的基因为基础的治疗人类OA。
Gene therapy is an attractive strategy for the durable treatment of human osteoarthritis (OA), a gradual, irreversible joint disease. Gene carriers based on the small human adeno-associated virus (AAV) exhibit major efficacy in modifying damaged human articular cartilage in situ over extended periods of time. Yet, clinical application of recombinant AAV (rAAV) vectors remains complicated by the presence of neutralizing antibodies against viral capsid elements in a majority of patients. The goal of this study was to evaluate the feasibility of delivering rAAV vectors to human OA chondrocytes in vitro and in an experimental model of osteochondral defect via polymeric micelles to protect gene transfer from experimental neutralization. Interaction of rAAV with micelles of linear (poloxamer PF68) or X-shaped (poloxamine T908) poly(ethylene oxide) (PEO) and poly(propylene oxide) (PPO) copolymers (PEO-PPO-PEO micelles) was characterized by means of isothermal titration calorimetry. Micelle encapsulation allowed an increase in both the stability and bioactivity of rAAV vectors and promoted higher levels of safe transgene (lacZ) expression both in vitro and in experimental osteochondral defects compared with that of free vector treatment without detrimental effects on the biological activity of the cells or their phenotype. Remarkably, protection against antibody neutralization was also afforded when delivering rAAV via PEO-PPO-PEO micelles in all systems evaluated, especially when using T908. Altogether, these findings show the potential of PEO-PPO-PEO micelles as effective tools to improve current gene-based treatments for human OA.