Enhanced Chondrogenic Differentiation Activities in Human Bone Marrow Aspirates via sox9 Overexpression Mediated by pNaSS-Grafted PCL Film-Guided rAAV Gene Transfer
Enhanced Chondrogenic Differentiation Activities in Human Bone Marrow Aspirates via sox9 Overexpression Mediated by pNaSS-Grafted PCL Film-Guided rAAV Gene Transfer
复制标题
DOI：
10.3390/pharmaceutics12030280
复制
发表时间：
2020-03
期刊：
影响因子：
5.4
通讯作者：
J. Venkatesan;Weikun Meng;A. Rey-Rico;G. Schmitt;S. Speicher-Mentges;C. Falentin-Daudré;A. Leroux;H. Madry;V. Migonney;M. Cucchiarini
中科院分区：
文献类型：
--
作者：
J. Venkatesan;Weikun Meng;A. Rey-Rico;G. Schmitt;S. Speicher-Mentges;C. Falentin-Daudré;A. Leroux;H. Madry;V. Migonney;M. Cucchiarini
背景资料：使用非侵入性支架引导的基因治疗程序在关节软骨病变部位递送治疗基因是一种有前途的方法，可以刺激软骨修复，同时保护货物免受有害的免疫反应，特别是当靶向天然微环境中的软骨修复骨髓源性间充质基质细胞时，如骨髓抽吸物。研究方法：在这里，我们评估了通过重组腺相关病毒（rAAV）载体向人骨髓抽吸物提供软骨特异性性别决定区Y型高迁移率族蛋白9（SOX 9）转录因子序列的益处，所述重组腺相关病毒（rAAV）载体由聚（ε-己内酯）（PCL）膜递送，所述PCL膜通过与聚（苯乙烯磺酸钠）（pNaSS）接枝功能化以随时间推移增强骨髓软骨形成潜力。结果如下：相对于其他条件（没有载体涂层的pNaSS接枝和未接枝PCL膜），在用包被有候选rAAV-FLAG-hsox 9（携带人sox 9基因序列的FLAG标记的rAAV载体）载体的pNaSS接枝或未接枝PCL膜处理至少21天的抽吸物中观察到有效的sox 9过表达。通过rAAV sox 9/pNaSS移植或未移植的PCL膜过表达sox 9导致吸出物中生物学和软骨形成分化活性（基质沉积）增加，同时包含过早成骨和肥大而不影响细胞增殖，当使用pNaSS移植膜时注意到更有效的作用。结论：这些发现显示了通过PCL膜引导的治疗性rAAV（sox 9）递送靶向患者骨髓的益处，其作为用于在翻译应用中增强软骨修复的未来策略的现成系统。
Background: 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. Methods: 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. Results: Effective sox9 overexpression was observed in aspirates treated with pNaSS-grafted or ungrafted PCL films coated with the candidate rAAV-FLAG-hsox9 (FLAG-tagged rAAV vector carrying a human sox9 gene sequence) vector for at least 21 days relative to other conditions (pNaSS-grafted and ungrafted PCL films without vector coating). Overexpression of sox9 via rAAV sox9/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. Conclusions: These findings show the benefits of targeting patients’ bone marrow via PCL film-guided therapeutic rAAV (sox9) delivery as an off-the-shelf system for future strategies to enhance cartilage repair in translational applications.