In situ Cell Engineering for On-demand TIMP Expression in Osteoarthritis

骨关节炎（OA）是一种常见的退行性关节疾病，也是导致残疾的主要原因。现时 没有疾病修饰骨关节炎药物（DMOAD）。诊断和药物干预发生 大多数是晚期，目前的治疗方法只能在疾病进展前提供暂时的姑息性缓解 需要进行关节置换。OA的患病率很高，在40岁以上的人群中约占27%， 创伤后OA（PTOA）的发生率甚至更高（超过50%）。给定 由于其高发病率和可预测性，PTOA有可能被治疗，这是一种既 有利于实现改善疾病的结局，并且在商业/临床上可行，提供治疗 提供长期保护。我们的目标是通过原位永久转换细胞来实现持久的关节保护 进入“按需”TIMP-3“工厂”，利用TIMP-3作为泛MMP抑制剂， OA病理学，包括软骨降解、血管生成和炎症。“按需”表达 TIMP-3将通过劫持Mmp 13启动子的靶向和永久性基因插入来实现。这 这种方法是基于非病毒CRISPR纳米颗粒，只有在以下情况下才激活TIMP-3表达： 存在病理性（OA）刺激，从而最小化潜在的副作用。我们建议优化纳米颗粒 用于非病毒基因敲入的制剂，并定量TIMP-3敲入的体外和体内治疗功效。 vivo.这种疗法有可能避免大关节患者生活质量的重大损失 我们的团队在细胞内递送（杜瓦尔）、聚合物和 纳米颗粒化学（D 'Arcy）、基因组编辑和合成生物学（Brunger）和PTOA生物学（Hasty）。

Osteoarthritis (OA) is a prevalent degenerative joint disorder and the leading cause of disability. Presently, there are no disease-modifying osteoarthritis drugs (DMOADs). Diagnosis and pharmacological intervention occur mostly at a late stage, and current treatments offer only temporary, palliative relief before disease progression necessitates joint replacement. OA prevalence is high, at roughly 27% of those over 40 years old, and occurrence of post-traumatic OA (PTOA) is even higher (over 50%) following injury of large joints such as the knee. Given its high incidence and predictability, PTOA has potential to be treated prophylactically, a strategy that is both conducive to achieving disease-modifying outcomes and commercially/clinically feasible, provided treatment offers long-term protection. We aim to achieve persistent joint protection by permanently converting cells in situ into “on-demand” TIMP-3 “factories”, harnessing TIMP-3 as a pan-MMP inhibitor that blocks multiple aspects of OA pathology, including cartilage degradation, angiogenesis, and inflammation. “On-demand” expression of TIMP-3 will be achieved via a targeted and permanent gene insertion that hijacks the Mmp13 promoter. This approach is based on a nonviral CRISPR-based nanoparticle and activates TIMP-3 expression only when pathological (OA) stimuli are present, minimizing potential side-effects. We propose to optimize a nanoparticle formulation for non-viral gene knock-in and quantify the therapeutic efficacy of TIMP-3 knock-in in vitro and in vivo. This therapy has potential to avoid significant loss in quality of life for patients who experience a large joint injury and is uniquely enabled by our team with expertise in intracellular delivery (Duvall), polymer and nanoparticle chemistry (D’Arcy), genome editing and synthetic biology (Brunger), and PTOA biology (Hasty).