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.

项目摘要 我们的长期目标是设计可以报告和治疗早期1型的治疗性免疫细胞 糖尿病（T1D）。理想情况下，这些细胞应归还胰岛，检测和整合早期的多因素特征 阶段疾病，以响应诱导局部免疫抑制以阻断β细胞的破坏。虽然 这似乎是一个非常困难的多层次挑战，在工程免疫方面取得了显着进步 识别和杀死癌症的细胞产生了广泛的新技术和方法 购买以对T1D的基于细胞的疗法进行。在这里，我们建议重新利用，重定向和扩展单元格 构建能够敏感性和治疗的合成免疫细胞（CD4+ T细胞）的工程方法 T1D等早期自身免疫性疾病。考虑到实现这一目标的挑战，我们建议 采用模块化方法 - 我们分解了理想的抗T1D细胞疗法必须实现的目标 三个不同的子任务。我们的重点将放在独立工程和验证电池电路模块上 可以实现这些子任务，然后可以将其链接在一起多种组合，以开发一个选项 全面疗法。这些模块化目标在我们的具体目的中概述了： 目标1。胰岛传感/靶向|识别和 在胰腺/小岛上建立居住/活动。胰腺/胰岛特异性抗原的传感器；胰岛 使用同步受体受到限制激活；和通过合成粘合蛋白进行的胰岛运输。 目标2。自身免疫性疾病感应|工程师合成免疫细胞感知并报告 与T1D发作相关的局部免疫扰动。检测存在的工程师传感器 自动反应性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：通过诱导局部保护胰岛的工程师治疗细胞 响应疾病感应的免疫抑制输出。工程师输出响应包括 诱导局部产生抑制性细胞因子（IL10，TGFB），炎性细胞因子下沉（CD25）和其他 Beta细胞保护因素。我们将测试从目标链接疾病传感电路的多种配置 AIM 3的抑制输出1和2。 为了开发用于治疗T1D的创新细胞工程平台，我们建议专注于人类多能 干细胞（HPSC）衍生的胰岛作为一种高度柔韧性系统，可以在其中评估免疫保护功能。 可以很容易地生成HPSC-Cells，并经过一定的修改，以添加传感器或 杀人，允许上述电路模块的快速设计构建迭代循环（即 使每个目标都非依赖于其他目标）。 HPSC胰岛也可以用于评估免疫保护 在体外和体内（小鼠植入）对各种胰岛细胞毒性T细胞的反应 肾胶囊），使该平台非常适合原则评估和细胞电路优化。