Abstract:
Osteoarthritis (OA) is a painful and debilitating disease affecting over 500 million people worldwide. Intraarticular injection of mesenchymal stromal cells (MSCs) shows promise for the clinical treatment of OA, but the lack of consistency in MSC preparation and application makes it difficult to further optimize MSC therapy and to properly evaluate the clinical outcomes. In this study, we used Sox9 activation and RelA inhibition, both mediated by the CRISPR-dCas9 technology simultaneously, to engineer MSCs with enhanced chondrogenic potential and downregulated inflammatory responses. We found that both Sox9 and RelA could be fine-tuned to the desired levels, which enhances the chondrogenic and immunomodulatory potentials of the cells. Intraarticular injection of modified cells significantly attenuated cartilage degradation and palliated OA pain compared with the injection of cell culture medium or unmodified cells. Mechanistically, the modified cells promoted the expression of factors beneficial to cartilage integrity, inhibited the production of catabolic enzymes in osteoarthritic joints, and suppressed immune cells. Interestingly, a substantial number of modified cells could survive in the cartilaginous tissues including articular cartilage and meniscus. Together, our results suggest that CRISPR-dCas9-based gene regulation is useful for optimizing MSC therapy for OA.
摘要:
骨关节炎(OA)是一种痛苦和衰弱的疾病,影响全球超过5亿人。间充质基质细胞(MSCs)的关节内注射显示出有望用于OA的临床治疗,但MSC制备和应用缺乏一致性,使得进一步优化MSC治疗和正确评估临床结局变得困难.在这项研究中,我们使用Sox9激活和RelA抑制,两者同时由CRISPR/dCas9技术介导,工程化具有增强的软骨形成潜力和下调的炎症反应的MSC。我们发现Sox9和RelA都可以微调到所需的水平,增强细胞的软骨形成和免疫调节潜能。关节内注射修饰细胞可显着减轻软骨降解和缓解OA疼痛,与注射细胞培养基或未修饰的细胞相比。机械上,修饰后的细胞促进了对软骨完整性有益的因子的表达,抑制骨关节炎关节中分解代谢酶的产生,抑制免疫细胞。有趣的是,大量的修饰细胞可以在软骨组织中存活,包括关节软骨和半月板。一起,我们的结果表明,基于CRISPR/dCas9的基因调控可用于优化MSC治疗OA.