Project 2 - Preclinical studies: Overcoming tumor heterogeneity

项目2 - 临床前研究：克服肿瘤异质性

• 批准号: 10241978
• 负责人: Steven Mark Albelda
• 金额: $ 51.54万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-14 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10241978
• 关键词: Address; Animals; Antibodies; Antigen Targeting; Antigens; Area; Biological Assay; Bystander Effect; CD19 Antigens; CTLA4 gene; Cancer Patient; Cells; Chimeric Proteins; Clinical; Clinical Trials; Clinical Trials Design; Combined Modality Therapy; Cross Presentation; Data; Defect; Dendritic Cells; Epitope spreading; Fibroblasts; Goals; Hematologic Neoplasms; Hematology; Heterogeneity; Heterophile Antigens; Human; Human Cell Line; Immune; Immunodeficient Mouse; Immunosuppression; Infrastructure; Knockout Mice; Ligands; Malignant Neoplasms; Malignant Pleural Mesothelioma; Malignant neoplasm of lung; Malignant neoplasm of ovary; Malignant neoplasm of pancreas; Malignant neoplasm of thorax; Mesothelioma; Monoclonal Antibody F19; Mus; Non-Small-Cell Lung Carcinoma; Patients; Process; Research Personnel; Safety; Sampling; Series; Solid Neoplasm; Surface Antigens; T cell therapy; T-Lymphocyte; Toxic effect; Translating; Treatment Efficacy; Tumor Antigens; Viral Antigens; cell type; chimeric antigen receptor; cytokine; efficacy evaluation; fetal liver kinase-2; fibroblast-activating factor; immune checkpoint; improved; inhibitor/antagonist; leukemia; leukemia treatment; mesothelin; mouse model; neoantigens; neoplastic cell; novel strategies; preclinical study; preclinical trial; programmed cell death protein 1; programs; safety and feasibility; success; trafficking; tumor; tumor growth; tumor heterogeneity; tumor microenvironment

Project 2. Preclinical Trials Abstract Non-small cell lung cancer (NSCLC) and malignant pleural mesothelioma (MPM) are highly lethal cancers with suboptimal therapies. The goal of this Program Project and of Project 2 is to successfully implement the use of T cells expressing chimeric antigen receptors (CARs), an approach that has been highly successful in leukemias. Our group was among the first to inititate CAR T cell trials focused on solid tumors by targeting the tumor-associated antigen mesothelin. However, despite showing safety and feasibility, therapeutic efficacy was limited. We hypothesize that one of the important issues limiting success in solid tumors is tumor antigen heterogeneity. Even at baseline, it is likely that not all tumor cells will express the targeted antigen. The primary focus of this Project will be to explore ways to overcome heterogeneity in solid tumor CAR T cell therapy. Our first approach will be to attack the tumor stroma. We previously showed that CAR T cells targeted to mouse fibroblast activation protein (FAP) on tumor fibroblasts can decrease the tumor matrix, augment endogenous intratumoral T cells, and slow tumor growth with minimal toxicity. In Aim 1, we propose to study a CAR targeted to human FAP that can be used in our clinical trial. In Aim 1A, we will assess the efficacy of the anti-human-CAR T cells using a new mouse model which combines human CAFs and human tumors in immunodeficient mice. In Aim 1B, we will evaluate the safety of anti-human FAP CAR T cells on primary human cell lines. In Aim 1C, we will evaluate combination therapies using the stromal-targeted CAR (FAP-CAR) with a tumor-cell targeted CAR (mesothelin-CAR). In Aim 2, we will explore the key issue of whether mesothelin and FAP CARs are able to induce bystander effects. In Aim 2A, we will study CAR- induced bystander effects by injecting mice with varying ratio's of tumor cells expressing or not expressing mesothelin and defining bystander effect as the minimal percentage of non-targeted tumor cells that still allow tumor elimination. Additional studies are proposed to determine the mechanisms by which the bystander effect occurs. In Aim 2B, the process of epitope spreading will be studied by determining the ability of Meso- and FAP-CARs to induce or stimulate endogenous T cells directed against tumors expressing a xenoantigen, a viral antigen, and a neoantigen. We will define the amount of baseline epitope spreading that is present using tetramer assays and a T cell cross presentation proliferation assay. In Aim 3, we will explore ways in which CAR-induced antigen spreading can be augmented by immune activators such as antibodies to PD1 and CTLA-4 or an IDO inhibitor (Aim 3A) and by genetically altering CAR T cells to produce FMS-like tyrosine kinase 3 ligand (FLT3L), a powerful inducer of dendritic cells (Aim 3B). Successful completion of these studies will provide key information for the field of adoptive transfer of T cells for solid tumors (especially with regard to the very understudied issue of overcoming tumor antigen heterogeneity) and provide important data to inform the designs of the clinical trials in Project 1. 1

项目2.临床前试验 摘要 非小细胞肺癌（NSCLC）和恶性胸膜间皮瘤（MPM）是高度致命的癌症， 次优疗法本方案项目和项目2的目标是成功实施 表达嵌合抗原受体（汽车）的T细胞，这种方法在治疗方面非常成功 白血病我们的团队是第一批启动CAR T细胞试验的团队之一，该试验主要针对实体瘤， 肿瘤相关抗原间皮素。然而，尽管显示出安全性和可行性， 是有限的。我们推测，限制实体瘤治疗成功的重要问题之一是肿瘤抗原。 异质性即使在基线，也可能不是所有肿瘤细胞都表达靶抗原。的 本项目的主要重点是探索克服实体瘤CAR T异质性的方法， 细胞疗法我们的第一步是攻击肿瘤间质。我们之前的研究表明， 靶向小鼠成纤维细胞活化蛋白（FAP）对肿瘤成纤维细胞的作用可以减少肿瘤基质， 增加内源性肿瘤内T细胞，并以最小的毒性减缓肿瘤生长。在目标1中，我们建议 研究一种靶向人类FAP的CAR，可以用于我们的临床试验。在目标1A中，我们将评估 使用新的小鼠模型的抗人CAR T细胞的功效，所述小鼠模型组合了人CAF和人CAR T细胞。 免疫缺陷小鼠的肿瘤。在目标1B中，我们将评估抗人FAP CAR T细胞对小鼠的安全性。 原代人类细胞系在目标1C中，我们将评估使用基质靶向CAR的联合治疗。 在一个实施方案中，本发明提供了一种具有肿瘤细胞靶向CAR（间皮素-CAR）的FAP-CAR。在目标2中，我们将探讨以下关键问题 间皮素和FAP汽车是否能够诱导旁观者效应。在Aim 2A中，我们将研究CAR- 通过给小鼠注射不同比例的表达或不表达的肿瘤细胞诱导旁观者效应， 间皮素和定义旁观者效应作为最小百分比的非靶向肿瘤细胞，仍然允许 肿瘤消除建议进行更多的研究以确定旁观者效应的机制。 发生。在目的2B中，将通过测定Meso-和Meso-的能力来研究表位扩展的过程。 FAP-CAR诱导或刺激针对表达异种抗原的肿瘤的内源性T细胞， 病毒抗原和新抗原。我们将定义基线表位扩展的量，其存在于使用 四聚体测定和T细胞交叉呈递增殖测定。在目标3中，我们将探讨如何 CAR诱导的抗原扩散可以通过免疫活化剂如针对PD 1和PD 2的抗体来增强。 CTLA-4或IDO抑制剂（Aim 3A），并通过遗传改变CAR T细胞以产生FMS样酪氨酸 激酶3配体（FLT 3L），一种强大的树突状细胞诱导剂（Aim 3B）。成功完成这些 研究将为实体瘤的T细胞过继转移领域提供关键信息（特别是与 关于克服肿瘤抗原异质性的研究不足的问题）并提供重要数据 为项目1中的临床试验设计提供信息。 1