Genetically-engineered stem cells for self-regulating arthritis therapy

摘要 超过五分之一的成年人会出现与关节炎相关的疾病，而且患病率还在增加。当前的方法 调节类风湿关节炎(RA)患者的内源性炎症介质 给予抗细胞因子生物药物时的严重不良反应(AEs)，如感染 以固定剂量持续服用。随着类风湿关节炎的严重程度随着时间的推移而波动，特定的治疗方法的发展 可以通过产生不同水平的内源性炎症介质来感知和响应的策略 相应地，适当水平的抗细胞因子药物代表了一种有吸引力的替代方法， 可减轻持续生物给药所致的不良反应。这个项目的目标是将基因用于 设计干细胞，创造生物人工植入物，用于生物药物输送，作为RA的治疗方法。通过 结合合成生物学和组织工程学的原理，我们将开发出对 特异性促炎细胞因子，如白介素1、白介素6和肿瘤坏死因子α 以反馈控制、自我调节和多元化的方式生产靶向抗细胞因子药物。一个 这项研究的主要重点是微调这些重新编程的抗炎细胞以增强 细胞给药对低水平全身炎症反应的敏感度和特异度。合成的 基因电路也将被引入到这些细胞中，以允许外源控制、可调节和可诱导 可以暂时或永久禁用抗细胞因子药物生产的安全开关。这些都是精心设计的 细胞将被包裹在以琼脂糖为基础的植入物中，这些植入物将被放置在由 实验性类风湿关节炎，这些方法的长期安全性和有效性将通过临床、 组织学、分子和疼痛/行为测试。这种“设计者”单元的创建提供了这样一种可能性 用于治疗慢性炎症性疾病的长期反馈受控给药。

Abstract Arthritis-related conditions occur in over 1 in 5 adults, and the prevalence is increasing. Current approaches to modulate endogenous inflammatory mediators in rheumatoid arthritis (RA) predispose patients to significant adverse effects (AEs), such as infection, when anti-cytokine biologic drugs are delivered continually at fixed doses. As the severity of RA fluctuates over time, development of specific therapeutic strategies that can sense and respond to varying levels of endogenous inflammatory mediators by producing correspondingly appropriate levels of anti-cytokine drugs represents an attractive alternative approach that may mitigate AEs induced by continuous biologic administration. The goal of this project is to used genetically engineered stem cells to create bioartificial implants for biologic drug delivery as a therapy for RA. By combining principles of synthetic biology and tissue engineering, we will develop stem cells that respond to specific pro-inflammatory cytokines such as interleukin-1, interleukin-6, and tumor necrosis factor alpha by producing targeted anti-cytokine drugs in a feedback-controlled, self-regulating, and multiplexed manner. A primary focus of this study is to fine-tune these reprogrammed anti-inflammatory cells to enhance the sensitivity and specificity of cell-based drug delivery in response to low-level systemic inflammation. Synthetic gene circuits will also be introduced in these cells to allow for exogenously-controlled tunable and inducible safety switches that can temporarily or permanently disable anti-cytokine drug production. These engineered cells will be encapsulated in agarose-based implants that will be placed subcutaneously in mice induced with experimental RA, and the long-term safety and efficacy of these approaches will be assessed using clinical, histologic, molecular, and pain/behavior testing. The creation of such “designer” cells provides the possibility for long-term, feedback-controlled drug delivery for the treatment of chronic inflammatory diseases.