From the Past to the Future: Chimeric Antigen Receptor T cells for Lymphoid Malignancies

摘要/摘要(项目1) 针对CD19蛋白(CART19)的嵌合抗原受体(CAR)T细胞已经使 治疗多种B细胞恶性肿瘤，某些类型的肿瘤完全缓解率高达97% 晚期白血病。这些反应中的许多都是持续的，但CAR T细胞的增殖和持久性很差 输液后和抗原阴性逃逸是治疗失败的常见机制。这是至关重要的 研究在有反应的患者中驱动CAR T细胞功能成功的因素。先前的研究表明 在淋巴系统恶性肿瘤中，持久缓解与特定T细胞通路的激活和 只有部分患者经历了CAR T细胞扩增和抗肿瘤活性的治疗水平。在.期间 在之前的颁奖期间，我们成功地全面了解了T细胞的内在和- 耐药的外在机制以及导致复发的肿瘤细胞内在机制。在这里我们 提出广泛的下一代分析，以了解最佳CAR T细胞疗法的特性，阐明 耐药机制，并制定策略以克服耐药性并增强CAR T细胞活性 治愈更多血液病患者。我们将利用已经建立的成功 与拥有细胞制造、基因编辑和最新技术的核心实验室合作 与艺术相关的科学平台探索了一些创新目标。在目标1中，我们将开展广泛的 流式细胞仪分析T细胞受体(TCR)重排、载体拷贝数、T细胞亚群 细胞学、血清细胞因子和生物标志物(~7,000)，以及血液中CAR T细胞的转录图谱 以及肿瘤组织，并制定连接相关数据和结果的多组学模型。在目标2中，我们有 设计了一项高影响力的临床试验，以敲除CD5，一种非传统的负免疫检查点分子 和抗原受体信号通路的抑制剂，目标是增加3天的治疗指数 为复发/难治性B细胞恶性肿瘤制造CART19细胞。最后，在目标3中，我们将实施 增强CAR T细胞增殖、持久性和抗肿瘤作用基因的功能分析 为ALL、CLL和NHL开发“同类最佳”产品的功能。使用结构化的、多管齐下的战略， 我们希望改善基于CAR T细胞治疗B细胞恶性肿瘤的耐药性，并在临床上取得进展 几个下一代合成生物学工具。这项工作有望开辟这一领域的治疗视野 为癌症的过继免疫疗法提供了新的研究前景，可以转化为改进 其他癌症和自身免疫性疾病的治疗。

Summary/Abstract (PROJECT 1) Chimeric antigen receptor (CAR) T cells directed to the CD19 protein (CART19) have revolutionized the treatment of a variety of B cell malignancies, with complete remission rates as high as 97% in certain types of advanced leukemia. Many of these responses are sustained, but poor CAR T cell expansion and persistence following infusion and antigen-negative escape are common mechanisms of treatment failure. It is critical to investigate factors driving successful CAR T cell function in responding patients. Previous studies have indicated that in lymphoid malignancies, durable remission is associated with activation of specific T cell pathways and only some patients experience therapeutic levels of CAR T cell expansion and antitumor activity. During the previous award period, we successfully developed a comprehensive understanding of the T cell-intrinsic and - extrinsic mechanisms of resistance as well as tumor cell-intrinsic mechanisms that lead to relapse. Here we propose extensive next-generation analyses to understand properties of optimal CAR T cell therapy, elucidate mechanisms of resistance, and develop strategies to overcome resistance and enhance CAR T cell activity to cure more patients with hematologic diseases. We will take advantage of already established successful collaborations with core laboratories possessing expertise in cell manufacturing, gene editing, and state-of-the- art correlative science platforms to explore a number of innovative aims. In Aim 1, we will carry out an extensive analysis of T cell receptor (TCR) rearrangements, vector copy number, T cell subsets analyzed by flow cytometry, serum cytokines and biomarkers (~7,000), and transcriptional landscapes of CAR T cells in blood as well as tumor tissues and formulate multi-omics models linking correlative data and outcome. In Aim 2, we have designed a high-impact clinical trial to knockout CD5, an unconventional negative immune checkpoint molecule and inhibitor of antigen-receptor signaling, with the goal of increasing the therapeutic index of 3-day manufactured CART19 cells for relapsed/refractory B-cell malignancies. Finally, in Aim 3, we will carry out functional analyses of genes implicated in potentiating CAR T cell proliferation, persistence, and antitumor function to develop `best-in-class' products for ALL, CLL, and NHL. Using a structured, multi-pronged strategy, we hope to ameliorate resistance to CAR T cell-based therapies for B-cell malignancies and clinically advance several next-generation synthetic biology tools. This work is expected to open therapeutic horizons in the field of adoptive immunotherapy for cancer and offer new research prospects that could be translated to improving the treatment of other cancers and autoimmune disorders.