Development of antigen multimers for CAR T cell detection and functional profiling

开发用于 CAR T 细胞检测和功能分析的抗原多聚体

基本信息

批准号：
10741209
负责人：
Jun Huang
金额：
$ 23.91万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-09 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10741209
关键词：
Affinity Antigens Autologous Avidity B lymphoid malignancy B-Cell Acute Lymphoblastic Leukemia B-Cell Lymphomas Binding Biological Assay Biopsy Biopsy Specimen Blood Blood specimen CAR T cell therapy CD19 gene Cell Count Cell Separation Cell Therapy Cell physiology Cells Cellular biology Clinic Clinical Color Cytolysis Data Detection Development Drug Industry ERBB2 gene Engineering Epidermal Growth Factor Receptor Flow Cytometry Formulation Glycosides Immune response In complete remission Industry Infusion procedures Knowledge Label Lymphoma Malignant Neoplasms Mantle Cell Lymphoma Measures Mediating Molecular Monitor Monoclonal Antibodies Multiomic Data Multiple Myeloma Nature Patients Persons Phenotype Property Proteome Reagent Refractory Relapse Research Research Personnel Resolution Sampling Scientist Solid Neoplasm Sorting Specificity Stains Surface Antigens T cell differentiation Technology Testing Therapeutic Studies Time Treatment Efficacy Tumor Antigens Variant War antigen detection biobank cancer cell chimeric antigen receptor chimeric antigen receptor T cells clinical application cost cytokine epigenome experience experimental study flexibility immunoengineering improved in vivo large cell Diffuse non-Hodgkin&apos s lymphoma mesothelin monomer multiple omics patient population peripheral blood preclinical study profiles in patients receptor expression responders and non-responders response single cell analysis success tool transcriptome treatment response tumor weapons

项目摘要

PROJECT SUMMARY CAR T cell therapy offers new hope to patients with relapsed/refractory B cell malignancies and has become another weapon in the war on cancer. Despite the success and excitement around CAR therapies, sensitive and universal CAR-labeling reagents are required for accurate CAR T cell detection and profiling in research, industry, and in the clinic. To improve CAR T cell therapy efficacy, scientists, clinicians, and the pharmaceutical industry need accurate and multifunctional CAR-staining reagents to perform downstream assays, conduct preclinical studies, profile patient biospecimens, and develop new CARs. To overcome this technology gap, we developed antigen-multimers – high-avidity CAR detection reagents. Akin to conventional MHC multimers, our preliminary data demonstrate that antigen-multimers sensitively and specifically detect CD19-directed CAR T cells in commercial infusion products, in peripheral blood, and in tumor biopsies of diffuse large B cell lymphoma patients. Furthermore, we show that CD19-multimers can be readily employed to isolate CAR T cells from patient samples at different treatment stages for downstream flow cytometry and single-cell multi-omics analyses. In our application, based on our exciting preliminary observations, we propose to test the overarching hypothesis that antigen-multimers will enable CAR T cell detection and isolation and facilitate downstream single-cell analyses of CAR T cells from patient biospecimens. Our antigen-multimers can be used to generate new knowledge regarding CAR T cell function and in vivo efficacy. In Specific Aim 1, we propose to develop antigen-multimers for high-sensitivity CAR T cell detection and isolation. In addition to demonstrating the flexibility of our technology by generating 4 different CAR-detecting antigen-multimers, we will also develop approaches for simultaneously isolating and restimulating CAR T cells from patient samples in order to enable downstream phenotyping and functional analyses. In Specific Aim 2, we propose to develop antigen-multimer-enabled single CAR T cell multicolor flow cytometry and multi-omics. We will first use CD19-multimers to determine CAR expression, cell number, differentiation, functional phenotype, and persistence of CAR T cells by multicolor flow cytometry. We will next use CD19-multimers to isolate CAR T cells from responder and non-responder patients for subsequent single- cell multi-omics analyses. Leveraging the experience of two investigators with complementary expertise in immunoengineering (Huang) and translational/clinical CAR T cell therapy research (Kline), as well as the UChicago Cell Therapy Biobank which contains hundreds of lymphoma and myeloma patient samples treated with CD19- or BCMA-directed CAR T cell therapy, the proposed experiments will establish antigen-multimers as a new class of CAR-staining reagents with broad research and clinical applications.

项目概要 CAR T细胞疗法为复发/难治性B细胞恶性肿瘤患者带来了新的希望，并已成为尽管 CAR 疗法取得了成功并令人兴奋，但敏感和敏感的癌症领域的另一种武器。研究中准确的 CAR T 细胞检测和分析需要通用 CAR 标记试剂，为了提高 CAR T 细胞疗法的功效，科学家、超级明星和制药公司。行业需要准确且多功能的 CAR 染色试剂来执行下游分析、进行为了克服这一技术差距，我们进行临床前研究、分析患者生物样本并开发新的 CAR。开发了抗原多聚体——与传统 MHC 多聚体类似的高亲和力 CAR 检测试剂。初步数据表明抗原多聚体可灵敏且特异性地检测 CD19 定向的 CAR T 商业输液产品、外周血和弥漫性大 B 细胞淋巴瘤的肿瘤活检中的细胞此外，我们还发现 CD19 多聚体可轻松用于从患者体内分离 CAR T 细胞。在我们的下游流式细胞术和单细胞多组学分析中，不同处理阶段的样品。根据我们令人兴奋的初步观察，我们建议测试以下总体假设：抗原多聚体将使 CAR T 细胞检测和分离成为可能，并促进下游单细胞分析我们的抗原多聚体可用于产生新知识。关于 CAR T 细胞功能和体内功效。在具体目标 1 中，我们建议开发抗原多聚体用于高灵敏度 CAR T 细胞检测和除了通过生成 4 种不同的 CAR 检测来展示我们技术的灵活性之外。抗原多聚体，我们还将开发同时分离和重新刺激 CAR T 细胞的方法从患者样本中进行分析，以便进行下游表型分析和功能分析。在具体目标 2 中，我们建议开发抗原多聚体支持的单 CAR T 细胞多色流我们将首先使用 CD19 多聚体来确定 CAR 表达、细胞数量、接下来我们将通过多色流式细胞术分析 CAR T 细胞的分化、功能表型和持久性。使用 CD19 多聚体从有反应和无反应患者中分离 CAR T 细胞，用于后续单细胞治疗利用两位具有互补专业知识的研究人员的经验。免疫工程（Huang）和转化/临床 CAR T 细胞治疗研究（Kline），以及芝加哥大学细胞治疗生物库包含数百个接受治疗的淋巴瘤和骨髓瘤患者样本通过 CD19 或 BCMA 定向的 CAR T 细胞疗法，拟议的实验将建立抗原多聚体：具有广泛研究和临床应用的新型 CAR 染色试剂。