Chimeric Antigen Receptor T Cell Design Through Living Cell Systems Biology

项目摘要 嵌合抗原受体(CAR)T细胞代表了一种针对耐药癌症的令人兴奋的技术。 这些细胞已经被开发出来有效地靶向血液系统的恶性肿瘤，然而还没有发现 对抗实体肿瘤的临床成功，实体肿瘤是癌症死亡的主要原因。实体瘤有 无数种免疫抑制机制阻止了有效的CAR T细胞治疗，新的方法是 既需要阐明这些机制，也需要引导这些细胞的工程指向靶向 实体瘤。这项提案将采用综合生物信息学方法来确定佐剂靶标以 加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会 在这些位点的单细胞水平上进行测序，并与CAR T细胞治疗的疗效进行比较。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器 蛋白质将在培养中的CAR T细胞中动态成像，因为它们识别和靶向抗原产生 细胞。这些完全不同的数据集的计算集成将产生将被内置到 这辆车的设计。这些新结构将在体外和体内得到验证，并将为 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.