Mechanisms of self-assembly temperature-sensitive polymers forenhanced rAAV-mediated gene transfer into human mesenchymalstem cells

自组装温度敏感聚合物增强rAAV介导的基因转移至人间充质干细胞的机制

• 批准号: 288014206
• 负责人: Dr. Ana Rey Rico
• 金额: --
• 依托单位: Lehrstuhl für Experimentelle Orthopädie und Arthroseforschung
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/288014206?language=en
• 关键词: Mechanisms; self; assembly; temperature; sensitive

Damage to the articular cartilage is a prevalent, major unresolved problem in a variety of joint disorders including trauma and osteoarthritis (OA). Current therapeutic options to repair traumatic or OA cartilage are diverse, but none can fully restore the cartilage function. Gene therapy allows for a direct application of genes coding for therapeutic factors in sites of injury, resulting in a temporarily and spatially defined delivery of the candidate agent.Recombinant adeno-assciated viral vectors (rAAV) are particularly clinically adapted tools to achieve this goal.Important limitations to their use still include a relatively low efficacy of penetration (~ 13% of effective internalization upon vector-cell contact via the heparan sulphate proteoglycan cellular receptor), the presence of neutralizing antibodies against the viral capsid in individuals, and the possible presence of inhibitory anticoagulants (heparin) in samples.A potential approach to overcome these issues is to deliver rAAV via polymeric biomaterials in order to enhance the rate of vector transfer via different cell entry mechanisms.The goal of this project is to test the hypothesis that rAAV can be effectively encapsulated and released from micelles and hydrogels of poly-ethylene oxide (PEO) and poly-propylene oxide (PPO) triblock copolymers to target hMSCs and human articular chondrocytes as key cell populations involved in the processes of cartilage repair.The differential features (composition vs. structure) of these copolymers enables the obtaining of tunable rAAV delivery systems for achieving high and sustained levels of gene transfer.This strategy could constitute a new safe, effective tool for delivering of therapeutic candidates in localized sites of cartilage lesions as a means to improve the repair processes of this tissue.

关节软骨损伤是包括创伤和骨关节炎（OA）在内的各种关节疾病中普遍存在的主要未解决的问题。目前修复外伤性或OA软骨的治疗方案多种多样，但没有一种能完全恢复软骨功能。基因治疗允许在损伤部位直接应用基因编码治疗因子，导致候选药物的暂时和空间定义递送。重组腺相关病毒载体（rAAV）是实现这一目标的特别适用于临床的工具。其使用的重要限制仍然包括相对较低的渗透效率（通过硫酸肝素蛋白多糖细胞受体在载体细胞接触时有效内化约13%），个体中存在针对病毒衣壳的中和抗体，以及样品中可能存在抑制性抗凝剂（肝素）。克服这些问题的一个潜在方法是通过聚合生物材料递送rAAV，以便通过不同的细胞进入机制提高载体转移的速度。该项目的目标是测试rAAV可以有效地从聚环氧乙烷（PEO）和聚环氧丙烷（PPO）三嵌段共聚物的胶束和水凝胶中被封装和释放的假设，以靶向hMSCs和人关节软骨细胞作为参与软骨修复过程的关键细胞群。这些共聚物的不同特征（组成与结构）使得获得可调的rAAV递送系统，以实现高水平和持续的基因转移。这一策略可以构成一种新的安全、有效的工具，作为一种改善软骨组织修复过程的手段，在局部软骨病变部位提供治疗候选物。

项目成果

Supramolecular polypseudorotaxane gels for controlled delivery of rAAV vectors in human mesenchymal stem cells for regenerative medicine.

• DOI: 10.1016/j.ijpharm.2017.05.050
• 发表时间: 2017-10
• 期刊: International journal of pharmaceutics
• 影响因子: 5.8
• 作者: A. Rey-Rico;Heiko Babicz;H. Madry;A. Concheiro;C. Alvarez‐Lorenzo;M. Cucchiarini

PEO-PPO-PEO Carriers for rAAV-Mediated Transduction of Human Articular Chondrocytes in Vitro and in a Human Osteochondral Defect Model.

• DOI: 10.1021/acsami.6b06509
• 发表时间: 2016-08
• 期刊: ACS applied materials & interfaces
• 影响因子: 9.5
• 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