Venkatesan Jagadeesh K, Meng Weikun, Rey-Rico Ana, Schmitt Gertrud, Speicher-Mentges Susanne, Falentin-Daudré Céline, Leroux Amélie, Madry Henning, Migonney Véronique, Cucchiarini Magali
Center of Experimental Orthopaedics, Saarland University Medical Center, D-66421 Homburg/Saar, Germany.
LBPS/CSPBAT UMR CNRS 7244, Université Sorbonne Paris Nord, F-93430 Villetaneuse, France.
Pharmaceutics. 2020 Mar 21;12(3):280. doi: 10.3390/pharmaceutics12030280.
The delivery of therapeutic genes in sites of articular cartilage lesions using non-invasive, scaffold-guided gene therapy procedures is a promising approach to stimulate cartilage repair while protecting the cargos from detrimental immune responses, particularly when targeting chondroreparative bone marrow-derived mesenchymal stromal cells in a natural microenvironment like marrow aspirates.
Here, we evaluated the benefits of providing a sequence for the cartilage-specific sex-determining region Y-type high-mobility group box 9 (SOX9) transcription factor to human marrow aspirates via recombinant adeno-associated virus (rAAV) vectors delivered by poly(ε-caprolactone) (PCL) films functionalized via grafting with poly(sodium styrene sulfonate) (pNaSS) to enhance the marrow chondrogenic potential over time.
Effective overexpression was observed in aspirates treated with pNaSS-grafted or ungrafted PCL films coated with the candidate rAAV-FLAG-h (FLAG-tagged rAAV vector carrying a human gene sequence) vector for at least 21 days relative to other conditions (pNaSS-grafted and ungrafted PCL films without vector coating). Overexpression of via rAAV /pNaSS-grafted or ungrafted PCL films led to increased biological and chondrogenic differentiation activities (matrix deposition) in the aspirates while containing premature osteogenesis and hypertrophy without impacting cell proliferation, with more potent effects noted when using pNaSS-grafted films.
These findings show the benefits of targeting patients' bone marrow via PCL film-guided therapeutic rAAV () delivery as an off-the-shelf system for future strategies to enhance cartilage repair in translational applications.
使用非侵入性的、支架引导的基因治疗程序在关节软骨损伤部位递送治疗性基因，是一种有前景的方法，可刺激软骨修复，同时保护载体免受有害免疫反应的影响，特别是当在骨髓抽吸物等天然微环境中靶向软骨修复性骨髓间充质基质细胞时。
在此，我们评估了通过聚（ε-己内酯）（PCL）膜递送的重组腺相关病毒（rAAV）载体，向人骨髓抽吸物中提供软骨特异性性别决定区Y型高迁移率族框9（SOX9）转录因子序列的益处，该PCL膜通过与聚（苯乙烯磺酸钠）（pNaSS）接枝进行功能化，以随时间增强骨髓软骨生成潜力。
相对于其他条件（未涂覆载体的pNaSS接枝和未接枝的PCL膜），在用涂覆有候选rAAV-FLAG-h（携带人类基因序列的FLAG标签rAAV载体）载体的pNaSS接枝或未接枝的PCL膜处理的抽吸物中，观察到至少21天的有效过表达。通过rAAV/ pNaSS接枝或未接枝的PCL膜过表达导致抽吸物中的生物学和软骨分化活性（基质沉积）增加，同时包含过早的成骨和肥大，而不影响细胞增殖，使用pNaSS接枝膜时效果更显著。
这些发现表明，作为一种现成的系统，通过PCL膜引导的治疗性rAAV（）递送靶向患者骨髓，对未来在转化应用中增强软骨修复的策略有益。
