Project 3 - Immune analysis of clinical trial samples

项目3-临床试验样本的免疫分析

• 批准号: 10006193
• 负责人: Gerald P Linette
• 金额: $ 37.52万
• 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/10006193
• 关键词: Address; Aftercare; Animal Model; Antigen Targeting; Antigens; Area; B lymphoid malignancy; Bioinformatics; Biological Markers; Biometry; Biopsy; Biopsy Specimen; Blood; Blood specimen; CD19 gene; Cell Count; Cell Line; Cellular Immunity; Clinical; Clinical Data; Clinical Trials; Cyclophosphamide; Data; Ensure; Epitope spreading; Expression Profiling; Fibroblasts; Flow Cytometry; Genes; Genetic Transcription; Goals; Hematologic Neoplasms; Histologic; Human; Humoral Immunities; Immune; Immunity; Immunotherapy; In Situ Hybridization; Infusion procedures; Learning; Libraries; Link; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of ovary; Malignant neoplasm of pancreas; Malignant neoplasm of thorax; Mediating; Medicine; Mesothelioma; Methods; Modeling; Mus; Patients; Peptide Library; Peptides; Peripheral Blood Mononuclear Cell; Phage Display; Phenotype; Research Personnel; Sampling; Sampling Studies; Solid Neoplasm; Statistical Data Interpretation; T cell response; T cell therapy; T-Lymphocyte; Testing; Therapeutic; Tumor Antigens; adaptive immune response; antigen-specific T cells; breath composition; chimeric antigen receptor; clinical trial analysis; design; experimental study; fibroblast-activating factor; immunogenic; improved; leukemia; mesothelin; murine monoclonal antibody; neoantigens; next generation sequencing; peripheral blood; preclinical efficacy; preclinical study; programs; response; success; synthetic peptide; trafficking; tumor

Project 3. Immune Analysis of Clinical Trial Samples. Abstract The reasons for the lack of therapeutic success of CAR therapy for solid tumors, compared to that seen in hematologic malignancies, represents one of the most important questions in the field of immunotherapy. Programs that can integrate clinical trials (Project 1), preclinical studies (Project 2) and in-depth biocorrelate studies of samples derived from these human trials (this Project) are needed. As described in Project 1, our new clinical trial will use an optimized mesothelin targeted CAR, M5 huCART-meso, that has been constructed to contain a new anti-mesothelin scFv fragment that it is fully human (selected from a phage display library). M5 huCART-meso also had better preclinical efficacy than our previous SS1-mesothelin CAR (see Project 1). Our second planned clinical trial will use a CAR, huCART-FAP, directed to fibroblasts in tumor stroma by targeting fibroblast activation protein (FAP). The goal of Project 3 is to conduct biocorrelate studies of these clinical trials. We will focus on two main areas. In Aim 1, we will conduct studies to learn whether our CAR T cells in Project 1 are persisting and trafficking to tumors. We will analyze blood samples from our patients in Core B, however, we hypothesize that studying the ability of CARs to traffic to tumors is even more critical. CAR T cells isolated from tumor biopsies will be characterized for phenotype and function. In addition, the tumor will be examined for histological changes, target antigen expression, and transcriptionally profiled. Post-treatment biopsies will be expected from each patient in the trials proposed in Project 1 helping to ensure that materials from tumors are available for such studies. In Aims 2 and 3, we will conduct experiments aimed at studying epitope spreading which is a crucial, yet an understudied area of adoptive T cell transfer. In Aim 2, we will evaluate the hypothesis that the CAR T cells used in our clinical trials can enhanced/promote endogenous T cell immunity to shared tumor antigens (Aim 2A) and neoantigens (Aim 2B). In Aim 2A, overlapping peptide libraries from known tumor antigens as well as defined peptides will be used to interrogate PBMC for responses to shared tumor antigens. In Aim 2B, we will use next generation sequencing (NGS) of tumors to identify candidate neoepitopes. Tandem mini-gene constructs, as well as synthetic peptides encoding these neoepitopes, will be used to interrogate peripheral blood (and in limited cases, TIL) for new or enhanced T cell immunity after CAR therapy. Aim 2 in Project 3 is linked to Aim 2 in Project 2, where investigators will use animal models to look for the presence of antigen spreading and ways to augment this effect. Finally, in Aim 3 we will examine the hypothesis that CAR-mediated epitope spreading promotes a diverse repertoire of antigen-specific T cell responses in tumors by utilizing NSG methods to characterize TCR clonotypes in shared/neoantigen-specific T cell lines derived from blood. These TCR reference libraries will be employed to interrogate pre- and post- CAR T therapy in blood/tumor biopsy samples in order to characterize the intra-tumoral breath and composition of T cell immunity promoted by CAR-mediated epitope spreading.

项目 3. 临床试验样本的免疫分析。 抽象的 与所观察到的相比，CAR疗法对实体瘤缺乏治疗成功的原因 在血液系统恶性肿瘤中，这是免疫治疗领域最重要的问题之一。 可以整合临床试验（项目 1）、临床前研究（项目 2）和深入生物相关性的项目 需要对来自这些人体试验（本项目）的样本进行研究。正如项目 1 中所述，我们的 新的临床试验将使用优化的间皮素靶向 CAR，M5 huCART-meso，该药物已被 构建含有新的抗间皮素 scFv 片段，它是完全人源的（选自噬菌体） 显示库）。 M5 huCART-meso 也比我们之前的 SS1-间皮素 CAR 具有更好的临床前疗效 （参见项目1）。我们计划的第二个临床试验将使用 CAR，huCART-FAP，针对肿瘤中的成纤维细胞 通过靶向成纤维细胞激活蛋白（FAP）来抑制基质。项目3的目标是进行生物相关研究 这些临床试验。我们将重点关注两个主要领域。在目标 1 中，我们将进行研究以了解我们的 项目 1 中的 CAR T 细胞持续存在并运输至肿瘤。我们将分析我们的血液样本 然而，我们假设研究 CAR 运输至肿瘤的能力更加重要 批判的。从肿瘤活检中分离出的 CAR T 细胞将被表征为表型和功能。此外， 将检查肿瘤的组织学变化、靶抗原表达和转录谱。 在项目 1 中提出的试验中，预计对每位患者进行治疗后活检，以帮助确保 来自肿瘤的材料可用于此类研究。在目标 2 和 3 中，我们将进行旨在 研究表位扩散，这是过继性 T 细胞转移的一个重要但尚未得到充分研究的领域。在目标 2 中， 我们将评估临床试验中使用的 CAR T 细胞可以增强/促进的假设 对共享肿瘤抗原（目标 2A）和新抗原（目标 2B）的内源性 T 细胞免疫。在目标 2A 中， 来自已知肿瘤抗原以及定义的肽的重叠肽文库将用于询问 PBMC 对共同肿瘤抗原的反应。在目标 2B 中，我们将使用下一代测序 (NGS) 肿瘤以确定候选新表位。串联小基因构建体以及合成肽 编码这些新表位，将用于询问外周血（在有限的情况下，TIL）是否有新的或 CAR治疗后T细胞免疫力增强。项目 3 中的目标 2 链接到项目 2 中的目标 2，其中 研究人员将使用动物模型来寻找抗原传播的存在以及增强这种传播的方法 影响。最后，在目标 3 中，我们将检验 CAR 介导的表位扩散促进 利用 NSG 方法表征 TCR，研究肿瘤中抗原特异性 T 细胞反应的多样性 来自血液的共享/新抗原特异性 T 细胞系的克隆型。这些TCR参考库将是 用于检查血液/肿瘤活检样本中的 CAR T 治疗前和治疗后，以表征 CAR介导的表位扩散促进的肿瘤内呼吸和T细胞免疫的组成。