Synthetic circuits that drive infiltration of therapeutic T cells into immunologically cold tumors

驱动治疗性 T 细胞浸润至免疫冷肿瘤的合成回路

• 批准号: 10329255
• 负责人: Hana El-Samad
• 金额: $ 58.47万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-10 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10329255
• 关键词: Adhesions; Architecture; Behavior; Bilateral; Bone Marrow; Cell physiology; Cell-Cell Adhesion; Cells; Chemotaxis; Complex; Computer Models; Cytokine Signaling; Disease; Engineering; Goals; Hematopoietic Neoplasms; Home; Immune; Immunocompetent; Immunological Models; Immunologics; Immunotherapeutic agent; In Vitro; Infiltration; Lead; Libraries; Malignant Neoplasms; Malignant neoplasm of pancreas; Measures; Modeling; Mus; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Performance; Regulation; Signal Transduction; Solid; Solid Neoplasm; Spleen; T-Lymphocyte; Testing; Therapeutic; Tissue Model; Tissues; Tumor Antigens; Tumor Suppressor Proteins; Tumor-infiltrating immune cells; cell stroma; chimeric antigen receptor T cells; combinatorial; design; effective therapy; engineered T cells; exhaustion; experimental study; improved; in silico; in vivo; lymph nodes; mesothelin; migration; mouse model; neoplastic cell; novel therapeutics; pancreatic ductal adenocarcinoma model; programs; receptor; residence; response; single cell analysis; success; trafficking; tumor; tumor xenograft

Project summary/abstract Engineering trafficking circuits that drive therapeutic T cell infiltration into immune-excluded tumors Engineered chimeric antigen receptor (CAR) T cells have yet to achieve efficacy against solid cancers. Particularly challenging are immune-excluded “cold” tumors, which fail to accumulate large numbers of infiltrating T cells. In such cases, even if therapeutic T cells recognize and kill tumor cells in vitro, they will fail in vivo if they cannot infiltrate the tumor. We propose to engineer synthetic circuits that regulate T cell trafficking as a general strategy to drive therapeutic T cell infiltration into immunologically cold tumors. Immune cells naturally rely on complex trafficking behaviors. They patrol the body to surveil for diseases. Once diseased tissue is identified, they establish local residence and focally expand. Cell trafficking programs largely rely on regulation of three core cellular functions: 1) chemotaxis (modulating cell ingress and egress), 2) cell-cell adhesion (reducing cell egress), and 3) local proliferative signaling (cytokine signaling). While these mechanisms are naturally exploited by T cells, the evolved pathways are susceptible to suppression by numerous tumoral mechanisms. We hypothesize that synthetic regulatory circuits that directly wire tumor antigen signals to control therapeutic T cell chemotaxis, adhesion, and local proliferative signaling will improve targeted infiltration of immune excluded tumors. We propose to develop synthetic trafficking circuit designs through cycles of in silico modeling and in vitro experiments. We will test if synthetic trafficking circuits can improve CAR T cell efficacy, in vivo, using an immunocompetent murine model of immune-excluded pancreatic cancer. The resulting cell trafficking circuits should be applicable to a broad range of solid cancers, as well as other diseases. AIM 1. Design and characterize synthetic T cell trafficking circuits that coordinately regulate chemotaxis, adhesion and local proliferation in response to tumor antigen recognition 1.A. Use multi-scale computational modeling to explore design space of possible T cell trafficking circuits. Use model to identify circuit architectures and parameters that robustly increase tumor-selective infiltration 1.B. Construct a toolbox of modular trafficking circuits using synNotch receptors to control chemotaxis, adhesion, and proliferation in response to tumor antigen recognition; Construct combinatorial library of circuits. 1.C. Test synthetic trafficking circuits in vitro using multicompartment tissue models that measure T cell trafficking and migration. Evaluate circuits in vivo by measuring T cell trafficking in bilateral tumor xenograft mouse models. AIM 2. Use engineered trafficking circuits to improve anti-tumor efficacy in an immune excluded immunocompetent murine model of pancreatic ductal adenocarcinoma. Leverage synthetic trafficking circuits to improve murine α-Mesothelin CAR-T cell infiltration and clearance of KPC pancreatic ductal adenocarcinoma syngeneic mouse model. Use single cell analysis to assess impact on tumoral suppressor cells, stroma, host immune cell infiltration, and CAR T cell exhaustion.

项目概要/摘要 工程化运输电路，驱动治疗性T细胞浸润到免疫排斥肿瘤中 工程化嵌合抗原受体（CAR）T细胞尚未实现针对实体癌的功效。 特别具有挑战性的是免疫排斥的“冷”肿瘤，其不能积累大量的浸润性肿瘤细胞。 T细胞。在这种情况下，即使治疗性T细胞在体外识别并杀死肿瘤细胞，如果它们在体内不能识别并杀死肿瘤细胞， 无法渗透到肿瘤中我们建议设计调节T细胞运输的合成电路， 这是一种驱动治疗性T细胞浸润到免疫冷肿瘤中的一般策略。 免疫细胞自然依赖于复杂的运输行为。它们巡视身体以监测疾病。一旦 当病变组织被识别时，它们建立局部居住并局部扩张。细胞贩卖项目 依赖于三种核心细胞功能的调节：1）趋化性（调节细胞进入和外出），2）细胞-细胞 粘附（减少细胞外出）和3）局部增殖信号传导（细胞因子信号传导）。虽然这些机制 由于T细胞天然利用这些通路，因此进化的通路容易受到许多肿瘤抑制剂的抑制。 机制等我们假设，直接连接肿瘤抗原信号以控制 治疗性T细胞趋化性、粘附和局部增殖性信号传导将改善靶向浸润， 免疫排斥肿瘤。我们建议开发合成贩运电路设计，通过在硅片的循环 建模和体外实验。我们将测试合成运输回路是否可以提高CAR T细胞的功效， 体内，使用免疫排斥胰腺癌的免疫活性鼠模型。所得细胞 贩运线路应适用于广泛的实体癌以及其他疾病。 AIM 1.设计和表征协调调节趋化性的合成T细胞运输回路， 对肿瘤抗原识别应答的粘附和局部增殖 1.A.使用多尺度计算建模来探索可能的T细胞运输电路的设计空间。使用 模型，以识别电路结构和参数，这些结构和参数稳健地增加肿瘤选择性浸润 1.B.使用synNotch受体构建模块化运输电路工具箱，以控制趋化性，粘附， 以及对肿瘤抗原识别的增殖反应;构建组合电路库。 1.C.使用测量T细胞运输的多室组织模型在体外测试合成运输回路 和移民。通过测量双侧肿瘤异种移植小鼠模型中的T细胞运输来评估体内回路。 AIM 2.使用工程化的运输回路来改善免疫排斥的肿瘤中的抗肿瘤功效 胰腺导管腺癌免疫活性小鼠模型。 利用合成运输回路改善鼠α-间皮素CAR-T细胞浸润和清除 KPC胰腺导管腺癌同基因小鼠模型。使用单细胞分析评估对 肿瘤抑制细胞、基质、宿主免疫细胞浸润和CAR T细胞耗竭。