CRISPRa构建超活化M2巨噬细胞用于关节炎治疗

关节炎是一类影响人体关节，导致疼痛、肿胀、僵硬和关节功能受损 的炎症性疾病。最常见的关节炎类型是骨性关节炎（OA）和类风湿性关节炎（RA），滑膜炎症和关节损伤是这两类疾病的共同特征。巨噬细胞是滑膜炎和软骨破坏反应中最重要的细胞类型之一，对于OA 和RA 的发生和发展起到了关键作用。近年来，许多研究表明，M2 型巨噬细胞具有抗炎作用，并且在治疗炎症性疾病方面具有潜在的应用前景。然而，传统的方法往往无法实现足够高效的M2 型巨噬细胞治疗效果。因此，在本研究中我们使用了CRISPRa 技术来激活M2 型巨噬细胞的IL-10 基因，构建了一种工程化的长期表达IL-10 的巨噬细胞（Elite-MΦ），并探索其在治疗OA 和RA 中的应用。实验结果表明，经过激活的巨噬细胞表达高水平的M2 标志物，同时可以产生更高水平的IL-10 等炎症抑制因子。在此基础上，我们研究了Elite-MΦ 在DMM 和CIA 两种实验性关节炎小鼠上的治疗效果。通过μCT扫描和组织学分析等方法我们发现，经过Elite-MΦ 治疗的关节炎小鼠骨关节结构保持完整，关节炎评分较低，而对照组小鼠则出现了明显的骨破坏和关节变形，进一步证明了Elite-MΦ 的治疗效果，为巨噬细胞用于关节炎治疗提供了潜在方案。

Arthritis is an inflammatory disease that affects human joints, causing pain, swelling, stiffness, and impaired joint function. The most common types of arthritis are osteoarthritis (OA) and rheumatoid arthritis (RA), which share synovial inflammation and joint damage as common features. Macrophages are one of the most important cell types in synovial inflammation and cartilage destruction reactions, and they play a critical role in the occurrence and development of OA and RA. In recent years, numerous studies have shown that M2 macrophages have anti-inflammatory effects and have potential applications in the treatment of inflammatory diseases. However, traditional methods often fail to achieve sufficiently effective therapeutic outcomes of M2 macrophages.
Therefore, in this study, we used CRISPRa technology to activate the IL-10 gene in macrophages and constructed an engineered lastingly IL-10 expressed macrophage (Elite-MΦ) for exploring its application in the treatment of OA and RA. The experimental results showed that the Elite-MΦ expressed high levels of M2 markers and produced higher levels of inflammation inhibitory factors such as IL-10. Based on this, we investigated the therapeutic effects of Elite-MΦ on experimental joint arthritis mice, including DMM and CIA models. Using methods such as micro-CT scanning and histological analysis, we found that joint arthritis mice treated with Elite-MΦ maintained intact bone joint structures and
had lower arthritis scores, while the control group mice showed obvious bone destruction and joint deformities, further demonstrating the  therapeutic effects of Elite-MΦ and providing potential strategies for macrophage-based arthritis treatment.

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