Precise tumor targeting with logic CAR circuits

项目摘要 这项建议描述了一种独特的人T细胞超CAR系统，以解决体内的非肿瘤毒性-- TY，使用已建立的肝脏转基因表达小鼠模型。我们将使用我们最近开发的裂解装置， 通用和可编程(以上)车载系统，同时包含多个关键的UP-UP 等级，例如微调T细胞激活强度的能力，感知和逻辑响应多个 抗原，并独立地同时调节两个T细胞亚型。具体地说，我们将评估两个 电路类型：(1)和(2)NIMPLY(A和NOT B)逻辑汽车电路，已确定 生物信息学是区分肿瘤和健康细胞的最佳电路类型。建议的经验如下： 实验将检验这样的假设：(1)我们的模块化和可调逻辑CAR T系统是唯一的 适合于在体内防止靶点上的肿瘤外毒性，即使当健康组织表达焦油- GET抗原在肿瘤附近，(2)我们的CAR系统的肿瘤特异性和活性是 通过受控交付而得到加强。重要的是，初步数据支持这些假设，而且- 在这项建议中，建立了表征材料和严格的实验设计，并具有基本的 涵盖合成生物学、免疫学、动物肿瘤的跨学科合作和专业知识 生物学和生物材料。这一五年计划的具体目标如下：目标1评估和记录- IC在体内的特异性高于CAR。目的2研究NIMPLY LOGIC的肿瘤特异性和毒性 车在活体内。目的3确定接头蛋白递送机制对肿瘤靶向性的影响。 这项工作的成果将建立逻辑CAR系统的临床潜力，并将它们的使用扩展到ADI。 传统癌症，包括实体瘤。

Project Summary This proposal describes a unique SUPRA CAR system in human T cells to address in vivo off-tumor toxici- ty, using an established liver transgene expression mouse model. We will use our recently developed split, universal and programmable (SUPRA) CAR system that simultaneously encompasses multiple critical up- grades, such as the ability to finely tune T cell activation strength, sense and logically respond to multiple antigens, and independently regulate two T cell subtypes simultaneously. Specifically, we will evaluate two types of circuits: (1) AND and (2) NIMPLY (A AND NOT B) logic CAR circuits, which have been determined bioinformatically as the best type for circuits for discriminating tumors from healthy cells. The proposed exper- iments will test the hypotheses that (1) our modular and tunable logic CAR T systems are uniquely suited for preventing on-target off-tumor toxicity in vivo, even when healthy tissues expressing a tar- get antigen are near the tumor, and (2) the tumor specificity and activity of our SUPRA CAR systems is enhanced by controlled delivery. Importantly, preliminary data support these hypotheses, and well- characterized materials and rigorous experimental designs are established in this proposal with essential cross-disciplinary collaborations and expertise that encompass synthetic biology, immunology, animal tumor biology, and biomaterials. The specific aims of this five-year proposal are as follows: Aim 1 evaluates AND log- ic SUPRA CAR specificity in vivo. Aim 2 investigate the tumor specificity and toxicity of NIMPLY logic SUPRA CAR in vivo. Aim 3 determine the effect of adaptor protein delivery mechanism on tumor-targeting specificity. Outcomes from this work will establish the clinical potential of logic CAR systems and expand their use to addi- tional cancers, including solid tumors.