Programmed cells for targeted articular regenerative medicine

项目总结 骨性关节炎(OA)影响着3200多万美国人，是全世界因残疾而失去生命的主要原因。 骨性关节炎的特征是关节软骨中富含II型胶原的基质分解和炎症。 指关节组织，如滑膜。目前还没有治疗关节炎的治疗疾病的药物，手术只是 进展性骨性关节炎患者的最终结果。虽然现有的手术方案提供了一些临床上的好处， 随着时间的推移，结果会下降，因为这些方法无法诱导强劲、持续的 关节组织。细胞工程策略已经被开发出来，以克服外科治疗的局限性。 这些策略部署细胞或基因递送载体，以向 然而，这些依赖于不受调控或仅受一般控制的转基因表达，这些转基因可能 长时间表达或在病理靶组织外表达会对健康产生负面影响。在这里，我们 建议利用合成生物学工具将转基因的表达限制在具有关节特征的位置 退化。我们的方法建立在我们使用定制的细胞感知和响应平台的基础上 工程学。这个人造细胞信号系统使我们能够对细胞进行编程，使其对选定的功能做出反应 通过执行已定义的转录程序来实现微环境。我们之前的研究表明 暴露的II型胶原蛋白是骨性关节炎的诊断标志。这项建议充分利用了我们最近的演示。 合成受体平台选择性地许可细胞检测II型胶原，然后上调 所选转基因的表达。我们工作的总体目标是建立合成受体控制的细胞 作为协调软骨修复和对抗与关节创伤或慢性炎症相关的炎症的媒介 关节发炎。这个项目将描述我们的受体驱动干细胞中介软骨的能力。 关节病体外模型的修复(目标1)，并将评估移植的SynNotch编程的能力 在创伤后骨性关节炎的活体模型中检测软骨细胞并对其作出反应(目标2)。

PROJECT SUMMARY Osteoarthritis (OA) affects over 32 million Americans and is a leading cause of years lost to disability worldwide. OA is characterized by the breakdown of the collagen II-rich matrix of articular cartilage, as well as inflammation of joint tissues such as the synovium. No disease-modifying drugs exist to treat arthritis, leaving surgery as the eventual result for patients with progressive OA. While existing surgical options offer some clinical benefit, outcomes decline over time due to the inability of these approaches to induce robust, sustained regeneration of joint tissues. Cell engineering strategies have been developed to overcome the limitations of surgical treatments. These strategies deploy cells or gene delivery vehicles to supply anabolic and/or anti-inflammatory factors to the joint; however, these rely on unregulated or only generically controlled expression of transgenes that may negatively impact health when expressed for long durations or outside of pathologic target tissues. Here, we propose to leverage synthetic biology tools to confine expression of transgenes to sites characterized by joint degeneration. Our approach builds on our use of a customized cell sense and response platform in regenerative engineering. This artificial cell signaling system enables us to program cells to react to selected features of a microenvironment by implementing defined transcriptional programs. Our prior studies have illustrated that exposed collagen II serves as a diagnostic hallmark of OA. This proposal capitalizes on our recent demonstration that the synthetic receptor platform selectively licenses cells to detect type II collagen and then upregulate expression of chosen transgenes. The overall goal of our work is to establish synthetic receptor-controlled cells as agents to orchestrate cartilage repair and combat the inflammation associated with joint trauma or chronic joint inflammation. This project will characterize the ability of our receptors to drive stem cells to mediate cartilage repair in an in vitro model of arthropathy (Aim 1) and will assess the ability of transplanted synNotch-programmed cells to detect and respond to cartilage degeneration in an in vivo model of post-traumatic OA (Aim 2).