Optimizing Dual-Targeted and Dual-Armored CAR T Cells for Small Cell LungCancer

摘要 患者自身的T细胞可以使用基因治疗技术进行修饰以表达受体，称为嵌合T细胞。 抗原受体或汽车，允许这些免疫T细胞识别肿瘤细胞表面的蛋白质， 反过来又允许这些CAR修饰的T细胞识别并杀死患者自身的肿瘤细胞。这种方法 在一些血液恶性肿瘤中已经成功，然而，迄今为止在固体中还没有成功。 肿瘤，包括小细胞肺癌（SCLC）。SCLC可能逃避T细胞介导的两种机制 杀伤是抗原表达的丧失和肿瘤微环境中T细胞功能的抑制。在 在这项提议中，我们将试图通过设计靶向两种SCLC抗原的CAR T细胞来克服这些障碍。 同时，其产生增强实体瘤中T细胞活性的多种因子（“盔甲”）。我们 假设这些双装甲、双靶向（DADT）CAR T细胞将更有效地对抗SCLC 比以前的T细胞介导的和免疫疗法。我们之前已经证明，CAR T细胞靶向 抗原GD 3或抗原DLL 3，两者都在大多数小细胞肺 癌症，能够在临床前系统中杀死SCLC细胞。此外，我们还开发了多种 装甲CAR T细胞，分泌IL-18或抗体衍生的单链可变片段等因子 阻断免疫检查点受体PD-1的scFv或阻断吞噬抑制信号的scFv 肿瘤细胞上的CD 47。所有这些装甲通过以下方式增强我们体内模型系统中的CAR T细胞活性： 不同的机制。在本提案的目标1中，我们将产生靶向DLL 3和GD 3的CAR T细胞， 同时，为了克服肿瘤中的抗原异质性和抗原丢失， 细胞介导的杀伤同时，在目标2中，我们将测试成对的装甲，以确定最多的一对 在体内有效增强单抗原靶向的CAR T细胞针对SCLC的活性， 免疫活性系统然后，我们分析了SCLC肿瘤微环境中的免疫细胞， CAR T细胞治疗以评估由装甲CAR T细胞介导的变化。在目标3中，我们将 联合收割机将这些方法结合起来，产生识别GD 3和DLL 3并产生多种装甲的CAR T细胞。 这些用于SCLC的DADT CAR T细胞可能适用于进一步的临床前测试，为临床试验做准备。 超出了本提案的范围，代表了SCLC的新治疗方法。鉴于我们的强大追踪 在CAR T细胞临床转化方面，我们完全期待有新的CAR T细胞适用于临床试验， 资金的终结。此外，这些新的CAR T细胞可用作探索相互作用的工具。 T细胞和SCLC微环境之间的关系。致小细胞肺癌的变化分析 本文提出的装甲CAR T细胞可能揭示了SCLC生物学的新方面，并阐明了 免疫逃逸和治疗失败。

ABSTRACT A patient's own T cells can be modified using gene therapy technology to express receptors, termed chimeric antigen receptors or CARs, which allow these immune T cells to recognize proteins on the tumor cell surface, and in turn allow these CAR modified T cells to recognize and kill the patient's own tumor cells. This approach has been successful in some hematological malignancies, however, it has not been successful to date in solid tumors including small cell lung cancer (SCLC). Two mechanisms by which SCLC may evade T cell-mediated killing are loss of expression of antigens, and suppression of T cell function in the tumor microenvironment. In this proposal, we will attempt to overcome these barriers by designing CAR T cells that target two SCLC antigens simultaneously, and that produce multiple factors (“armors”) that enhance T cell activity in solid tumors. We hypothesize that these dual-armored, dual targeted (DADT) CAR T cells will be more effective against SCLC than previous T cell-mediated and immune therapies. We have previously shown that CAR T cells targeted to either the antigen GD3 or to the antigen DLL3, both of which are expressed on the majority of small cell lung cancers, are capable of killing SCLC cells in preclinical systems. Additionally, we have developed multiple armored CAR T cells that secrete factors such as IL-18, or an antibody-derived single-chain variable fragment (scFv) that blocks the immune checkpoint receptor PD-1, or an scFv blocking the phagocytosis-inhibitory signal CD47 on tumor cells. All of these armors enhance CAR T cell activity in our in vivo model systems through different mechanisms. In Aim 1 of this proposal, we will generate CAR T cells targeting DLL3 and GD3 simultaneously, to overcome antigen heterogeneity and antigen loss in tumors as a means of escape from T cell-mediated killing. Simultaneously, in Aim 2, we will test pairs of armors to identify the pair that is the most effective at enhancing the activity of single antigen-targeted CAR T cells against SCLC in vivo in immunocompetent systems. We then analyze the immune cells in the SCLC tumor microenvironment following CAR T cell treatment to assess changes mediated by the armored CAR T cells. Ultimately, in Aim 3, we will combine these approaches to generate CAR T cells that recognize GD3 and DLL3 and produce multiple armors. These DADT CAR T cells for SCLC may be suitable for further preclinical testing in preparation for clinical trials beyond the scope of this proposal, representing a novel therapeutic approach to SCLC. Given our robust track record in CAR T cell clinical translation, we fully anticipate having new CAR T cells suitable for clinical trials at the conclusion of funding. Additionally, these novel CAR T cells may be used as tools to explore the interactions between T cells and the SCLC microenvironment. The analysis of changes in SCLC tumors induced by the armored CAR T cells proposed here may reveal novel aspects of SCLC biology and illuminate mechanisms of immune escape and treatment failure in SCLC.