Programmed cells for targeted articular regenerative medicine

项目摘要 骨关节炎（OA）影响着超过3200万美国人，是全世界因残疾而损失多年的主要原因。 OA的特征是关节软骨中富含II型胶原的基质的分解以及炎症 关节组织如滑膜。没有疾病修饰药物存在治疗关节炎，离开手术作为 对于患有进行性OA的患者的最终结果。虽然现有的手术选择提供了一些临床益处， 结果随着时间的推移而下降，因为这些方法无法诱导强大的，持续的再生， 关节组织已经开发了细胞工程策略来克服手术治疗的局限性。 这些策略部署细胞或基因递送载体以向受试者提供合成代谢和/或抗炎因子。 然而，这些依赖于转基因的不受调控的或仅受一般控制的表达， 当长时间表达或在病理靶组织外表达时，对健康产生负面影响。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以联合表达为特征的位点， 退化我们的方法建立在我们使用定制的细胞感觉和反应平台， 工程.这种人工细胞信号系统使我们能够对细胞进行编程，使其对细胞的特定特征做出反应。 微环境，通过执行定义的转录程序。我们之前的研究表明， 暴露的II型胶原蛋白是OA的诊断标志。这项建议利用了我们最近的演示 合成受体平台选择性地允许细胞检测II型胶原蛋白， 选择的转基因的表达。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试剂， 关节炎这个项目将表征我们的受体驱动干细胞介导软骨的能力 在关节病的体外模型中进行修复（目的1），并将评估移植的synNotch程序化的 细胞来检测和响应创伤后OA的体内模型中的软骨变性（目的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).