Genetically-engineered stem cells for self-regulating arthritis therapy

摘要 关节炎相关疾病发生在超过五分之一的成年人中，并且患病率正在增加。当前方法 调节类风湿性关节炎（RA）中的内源性炎症介质使患者易于 当输送抗细胞因子生物药物时，发生显著不良反应（AE），如感染 以固定剂量持续给药。由于RA的严重程度随时间而波动，因此开发特异性治疗药物可能是一个挑战。 这些策略可以通过产生不同水平的内源性炎症介质， 相应的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， 可减轻连续生物制剂给药引起的AE。这个项目的目标是利用基因 工程干细胞，以创造生物人工植入物的生物药物输送作为治疗RA。通过 结合合成生物学和组织工程学的原理，我们将开发出能够对 特异性促炎细胞因子如白细胞介素-1、白细胞介素-6和肿瘤坏死因子α， 以反馈控制、自我调节和多重方式产生靶向抗细胞因子药物。一 这项研究的主要焦点是微调这些重新编程的抗炎细胞，以增强免疫功能。 敏感性和特异性的细胞为基础的药物输送在低水平的全身炎症反应。合成 基因电路也将被引入这些细胞中，以允许外源控制的可调谐和可诱导的 可以暂时或永久禁用抗细胞因子药物生产的安全开关。这些工程化 将细胞包封在基于琼脂糖的植入物中， 实验性RA，这些方法的长期安全性和有效性将使用临床， 组织学、分子学和疼痛/行为测试。这种“设计者”细胞的创造提供了一种可能性， 用于治疗慢性炎症性疾病的长期反馈控制药物输送。

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.