Precise tumor targeting with logic CAR circuits

项目摘要 该提案描述了人T细胞中独特的SUPRA CAR系统，以解决体内肿瘤外毒性。 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分裂， 通用和可编程（SUPRA）CAR系统，同时包括多个关键的向上， 等级，如微调T细胞激活强度的能力，感觉和逻辑上响应于多个 抗原，并同时独立调节两种T细胞亚型。具体来说，我们将评估两个 电路类型：（1）AND（与）和（2）NIMOTH（A AND NOT B）逻辑CAR电路，已确定 作为区分肿瘤和健康细胞的电路的最佳类型。拟议的专家- 实验将检验以下假设：（1）我们的模块化和可调逻辑CAR T系统是唯一的 适用于预防体内靶向脱肿瘤毒性，即使健康组织表达焦油， 获得的抗原在肿瘤附近，和（2）我们的SUPRA CAR系统的肿瘤特异性和活性是 通过控制下交付得到加强。重要的是，初步数据支持这些假设，以及- 特征材料和严格的实验设计建立在这个建议与基本 跨学科合作和专业知识，包括合成生物学，免疫学，动物肿瘤， 生物学和生物材料。这一五年期提案的具体目标如下：目标1评估和记录- ic体内SUPRA CAR特异性。目的2研究尼莫地平SUPRA的肿瘤特异性和毒性 体内CAR。目的3探讨衔接蛋白传递机制对肿瘤靶向性的影响。 这项工作的结果将建立逻辑CAR系统的临床潜力，并将其应用扩展到增加 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.