Engineering synthetic immune cells with modular sentinel and therapeutic functions for T1D

工程合成免疫细胞具有模块化前哨和 T1D 治疗功能

• 批准号: 10594512
• 负责人: WENDELL A LIM
• 金额: $ 83.16万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10594512
• 关键词: Adhesions; Antigens; Autoimmune Diseases; Beta Cell; CD4 Positive T Lymphocytes; Cell Therapy; Cells; Cytoprotection; Cytotoxic T-Lymphocytes; Disease; Early Diagnosis; Engineering; Evaluation; Goals; Home; IL2RA gene; Immune; Immunosuppression; Immunotherapeutic agent; In Vitro; Inflammatory; Insulin-Dependent Diabetes Mellitus; Interleukin-10; Islet Cell; Kidney; Link; Malignant Neoplasms; Modeling; Mus; Output; Pancreas; Patients; Production; Proteins; Reporting; Sentinel; System; Testing; Therapeutic; Tissues; autoreactive T cell; capsule; cell type; cellular engineering; cytokine; design,build,test; flexibility; human pluripotent stem cell; immunoengineering; implantation; in vivo; innovation; insulin dependent diabetes mellitus onset; islet; islet stem cells; new technology; novel; novel strategies; protective factors; receptor; residence; response; sensor; synthetic construct; trafficking

PROJECT SUMMARY Our long-term goal is to engineer therapeutic immune cells that can report on and treat early-stage type 1 diabetes (T1D). Ideally, these cells should home to islets, detect and integrate multi-factor signatures of early- stage disease and, in response, induce localized immunosuppression to block destruction of beta cells. Although this seems like an extraordinarily difficult multi-level challenge, the remarkable progress in engineering immune cells to recognize and kill cancer has generated a broad set of new technologies and approaches that could be brought to bear on cell-based therapies for T1D. Here we propose to repurpose, redirect, and extend cell engineering approaches to construct synthetic immune cells (CD4+ T cells) capable of sensing and treating early-stage autoimmune disorders like T1D. Given the multiple challenges in achieving this goal, we propose to take a modular approach – we have broken up what an ideal anti-T1D cell therapy would have to achieve into three distinct subtasks. Our focus will be on independently engineering and validating cell circuit modules that can achieve these subtasks, which can then be linked together in multiple combinations to develop options for a full therapy. These modular objectives are outlined in our specific aims: Aim 1. ISLET SENSING/TARGETING | Engineer synthetic immune cell sentinels that recognize and establish residence/activity in the pancreas/islets. Sensors of pancreatic/islet specific antigens; islet restricted activation using synNotch receptors; and islet trafficking via synthetic adhesion proteins. Aim 2. AUTOIMMUNE DISEASE SENSING | Engineer synthetic immune cells that sense and report on local immune perturbations associated with T1D onset. Engineer sensors that detect presence of autoreactive T cells and elevated local levels of specific inflammatory cytokines Aim 3. IMMUNO-SUPPRESSIVE OUTPUT: Engineer therapeutic cells that protect islets by inducing local immunosuppressive outputs in response to disease sensing. Engineer output responses encompassing induced local production of suppressive cytokines (IL10, TGFb), inflammatory cytokine sinks (CD25), and other beta-cell protective factors. We will test multiple configurations of linking the disease sensing circuits from Aims 1 and 2 to the suppressive outputs from Aim 3. To develop innovative cell engineering platforms for treating T1D, we propose to focus on human pluripotent stem cell (hPSC)-derived islets as a highly flexible system in which to evaluate the immunoprotective function. hPSC-cells can be readily generated and genetically modified to add convenient model antigens for sensing or killing, allowing for the rapid design-build-test iterative cycles for the circuit modules described above (i.e., making each aim non-dependent on the others). hPSC islets can also be used to assess immunoprotective responses against a variety of islet-targeted cytotoxic T cells, both in vitro and in vivo (implantation in mouse kidney capsule), making this platform ideal for proof-of-principle evaluation and cell circuit optimization.

项目概要 我们的长期目标是设计能够报告和治疗早期 1 型疾病的治疗性免疫细胞 理想情况下，这些细胞应以胰岛为家，检测并整合早期糖尿病的多因素特征。 阶段疾病，并作为反应，诱导局部免疫抑制以阻止β细胞的破坏。 这似乎是一个极其困难的多层次挑战，免疫工程方面的显着进展 细胞识别和杀死癌症已经产生了一系列广泛的新技术和方法，这些技术和方法可以 在这里，我们建议重新利用、重新定向和扩展细胞。 构建能够感知和治疗的合成免疫细胞（CD4+ T 细胞）的工程方法 鉴于实现这一目标面临多重挑战，我们建议： 采用模块化方法——我们将理想的抗 T1D 细胞疗法必须实现的目标分解为 我们的重点将是独立设计和验证单元电路模块。 可以实现这些子任务，然后可以将这些子任务以多种组合方式链接在一起，以开发选项 我们的具体目标概述了这些模块化目标： 目标 1. 胰岛传感/靶向 | 设计可识别和定位的合成免疫细胞哨兵 建立胰腺/胰岛传感器的驻留/活性； 使用 synNotch 受体进行限制性激活；以及通过合成粘附蛋白进行胰岛运输。 目标 2. 自身免疫疾病传感 | 设计可感知并报告的合成免疫细胞 与 T1D 发病相关的局部免疫扰动设计传感器来检测是否存在。 自身反应性 T 细胞和特定炎症细胞因子的局部水平升高 目标 3. 免疫抑制输出：设计治疗细胞，通过诱导局部细胞来保护胰岛 响应疾病感知的免疫抑制输出。 诱导局部产生抑制性细胞因子（IL10、TGFb）、炎症细胞因子库（CD25）和其他 我们将测试连接 Aims 疾病传感电路的多种配置。 1 和 2 到目标 3 的抑制输出。 为了开发治疗 T1D 的创新细胞工程平台，我们建议重点关注人类多能细胞 干细胞 (hPSC) 衍生的胰岛作为高度灵活的系统来评估免疫保护功能。 hPSC 细胞可以很容易地生成并进行基因修饰，以添加方便的模型抗原以进行传感或 杀死，允许上述电路模块的快速设计-构建-测试迭代周期（即， 使每个目标不依赖于其他目标）也可用于评估免疫保护作用。 在体外和体内（植入小鼠体内）对多种胰岛靶向细胞毒性 T 细胞的反应 肾胶囊），使该平台成为原理验证评估和细胞电路优化的理想选择。