Chimeric Antigen Receptor T Cell Design Through Living Cell Systems Biology

项目摘要 嵌合抗原受体（CAR）T细胞代表了靶向耐药癌症的令人兴奋的技术。 这些细胞已被开发用于有效靶向血液恶性肿瘤，但尚未发现 在治疗实体瘤方面取得了临床成功，实体瘤占癌症死亡率的大部分。实体瘤有 无数的免疫抑制机制，阻止有效的CAR T细胞疗法，和新的方法， 需要阐明这些机制，并指导这些细胞的靶向工程， 实体瘤该提案将采用综合生物信息学方法来确定佐剂靶点， 增强实体瘤部位或早期转移部位的CAR T细胞治疗。基质细胞和免疫细胞 在这些位点以单细胞水平进行测序，并与CAR T细胞疗法的功效进行比较。这些 测序结果将指导活细胞成像实验，其中关键的转录因子和效应子 蛋白质将在培养的CAR T细胞中动态成像，因为它们识别并靶向抗原产生细胞， 细胞这些不同数据集的计算整合将产生辅助目标， 汽车设计。这些新的构建体将在体外和体内进行验证，并将为以下方面提供基础： 更先进的临床可用的CAR T细胞疗法。

Project Summary Chimeric antigen receptor (CAR) T cells represent an exciting technology for targeting drug-resistant cancers. These cells have been developed to effectively target hematological malignancies, however have yet to find clinical success against solid tumors, which account for the majority of cancer mortality. Solid tumors have myriad immunosuppressive mechanisms that prevent effective CAR T cell therapies, and new methods are needed to both elucidate these mechanisms as well as direct the engineering of these cells towards targeting solid tumors. This proposal will take an integrated bioinformatic approach to identifying adjuvant targets to enhance CAR T cell therapy at the site of a solid tumor or early metastatic site. Stromal and immune cells will be sequenced at the single cell level at these sites and compared to efficacy of CAR T cell therapy. These sequencing results will guide live cell imaging experiments, in which key transcription factors and effector proteins will be dynamically imaged in CAR T cells in culture as they recognize and target antigen-producing cells. Computational integration of these disparate datasets will result in adjuvant targets that will be built into the CAR design. These new constructs will be validated both in vitro and in vivo and will provide the basis for more advanced clinically available CAR T cell therapies.