Quantitative single-cell biomarkers of T-cells to optimize tumor immunotherapy

T 细胞的定量单细胞生物标志物可优化肿瘤免疫治疗

基本信息

批准号：
8413987
负责人：
Laurence J.N. Cooper
金额：
$ 72.91万
依托单位：
UNIVERSITY OF HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-19 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8413987
关键词：
Adoptive Immunotherapy Adoptive Transfer Allogenic Antibodies Antigen Receptors Antigen-Presenting Cells Applications Grants Archives Autologous B lymphoid malignancy Bioinformatics Biological Markers Blood Volume CD19 gene CD28 gene CD3 Antigens CD8B1 gene Cancer Center Cell Count Cell Therapy Cells Chronic Lymphocytic Leukemia Clinical Clinical Data Clinical Trials Correlative Study Cytolysis Data Development Event Fluorescence-Activated Cell Sorting Gene Expression Gene Expression Profile Generations HIV Hematologic Neoplasms Hematopoietic Stem Cell Transplantation Heterogeneity Human Image Analysis Immune Immune system Immunophenotyping Immunotherapy In Vitro Infusion procedures Institution MS4A1 gene Malignant Neoplasms Mediating Methodology Methods Molecular Profiling Monoclonal Antibodies Mus Outcome Patients Peripheral Blood Mononuclear Cell Phenotype Population Population Heterogeneity Preparation Production Refractory Regulatory T-Lymphocyte Research Personnel Resistance Retrieval Sample Size Sampling Screening procedure Signal Transduction Specificity T-Cell Receptor T-Lymphocyte Testing Therapeutic Toxic effect Translations Trees Tumor Antigens Universities Vaccines base cancer cell cellular imaging conventional therapy cytokine cytotoxicity density design effective therapy image processing improved in vivo killings leukemia/lymphoma neoplastic cell next generation pre-clinical response tool tumor

项目摘要

DESCRIPTION (provided by applicant): Immunotherapy infusing antibodies, vaccines, and cells is an effective treatment approach for human malignancies. Potent anti-tumor effects are realized by harnessing the specificity and associated effector functions of the immune system to recognize and eliminate cancer cells. Clinical trials have revealed the advantages of immune-based therapies, including (i) defined mechanisms of action, (ii) defined specificity and reduced deleterious off-target effects, (iii) lower toxicities than conventional approaches. Adoptive cell therapy (ACT), based on the adoptive transfer of T cells genetically modified to enforce expression of a chimeric antigen receptor (CAR), has shown considerable promise in clinical trials treating tumors refractory to all other treatment methods. In particular, the use of CAR+ T cells rendered specific for CD19 demonstrated significant anti-tumor effects in patients with CD19+ chronic lymphocytic leukemia (CLL) refractory to conventional therapies. These trials infuse a heterogeneous population of genetically modified T cells which have been propagated to clinically-sufficient numbers. While the therapeutic potential of infused T cells depends on their persistence, immunocorrelative studies evaluating survival of infused T cells are currently limited to (i) describing the immunophenotypes and function of whole populations of the T-cell inoculum and (ii) immunophenotypic analyses on T cells recovered after infusion. This grant application seeks to adapt single- cell imaging and retrieval to inform on the potency of clinical-grade CD19-specific CAR+ T cells by developing a unified platform for assessing both phenotype and function on the few T cells (and tumor cells) directly obtained (without in vitro manipulation) from recipients of immunotherapy. Our objective is to use high throughput single-cell nanowell screening (SNS) that we have developed to undertake an in-depth quantitative functional characterization (multiplexed cytokine secretion, phenotype, cytotoxicity, effect of regulatory T cells, Treg) of pre-infusion CAR+ T cells and compare these data to T cells recovered from the patient, post-infusion. These data will be used to (i) quantify the functionalit of infused T cells and the potential for anti-tumor effects and (ii) improve the generation of T cells for greater efficacy in next-generation clinical trials. We will validate our approach in Specific Aim 1 where we will define the integrated functional and molecular profiles of clinical- grade CD19-specific CAR+ T cells. In Specific Aim 2 we will implement our methodology to quantify the in vivo persistence of adoptively transferred cells. This will test the hypothesis tha SNS can quantify the therapeutic potential of clinical-grade T cells and in the formation of next-generation clinical trials. PUBLIC HEALTH RELEVANCE: Genetically modified T cells are being infused in clinical trials to target leukemia and lymphomas. This grant application validates tools for investigating the ability of the infused T cells to target tumor cells and applies this approach to improve our understanding of their therapeutic benefit in clinical trials. This will enable investigators to develop approaches to generating even more effective T cells to target hematologic malignancies in next-generation clinical trials.

描述（由申请人提供）：注入抗体、疫苗和细胞的免疫疗法是治疗人类恶性肿瘤的有效方法。通过利用免疫系统的特异性和相关效应功能来识别和消除癌细胞，从而实现有效的抗肿瘤作用。临床试验揭示了基于免疫的疗法的优点，包括（i）明确的作用机制，（ii）明确的特异性和减少有害的脱靶效应，（iii）比传统方法的毒性更低。过继细胞疗法 (ACT) 基于经过基因改造以强制表达嵌合抗原受体 (CAR) 的 T 细胞的过继转移，在治疗所有其他治疗方法都难治的肿瘤的临床试验中已显示出相当大的前景。特别是，使用对 CD19 具有特异性的 CAR+ T 细胞对传统疗法难治的 CD19+ 慢性淋巴细胞白血病 (CLL) 患者显示出显着的抗肿瘤作用。这些试验注入了异质的转基因 T 细胞群，这些细胞已经繁殖到临床上足够的数量。虽然输注 T 细胞的治疗潜力取决于其持久性，但评估输注 T 细胞存活率的免疫相关研究目前仅限于 (i) 描述 T 细胞接种物整个群体的免疫表型和功能，以及 (ii) 对 T 细胞的免疫表型分析。输注后细胞恢复。该拨款申请旨在通过开发一个统一的平台来评估直接获得的少数 T 细胞（和肿瘤细胞）的表型和功能，以适应单细胞成像和检索，以了解临床级 CD19 特异性 CAR+ T 细胞的效力。（未经体外操作）来自免疫治疗的接受者。我们的目标是利用我们开发的高通量单细胞纳米孔筛选 (SNS) 对输注前 CAR+ 进行深入的定量功能表征（多重细胞因子分泌、表型、细胞毒性、调节性 T 细胞、Treg 的作用） T 细胞并将这些数据与输注后从患者体内回收的 T 细胞进行比较。这些数据将用于 (i) 量化输注 T 细胞的功能和抗肿瘤作用的潜力，以及 (ii) 改善 T 细胞的生成，以便在下一代临床试验中发挥更大功效。我们将在具体目标 1 中验证我们的方法，其中我们将定义临床级 CD19 特异性 CAR+ T 细胞的综合功能和分子特征。在具体目标 2 中，我们将实施我们的方法来量化过继转移细胞的体内持久性。这将检验 SNS 可以量化临床级 T 细胞的治疗潜力并形成下一代临床试验的假设。公共健康相关性：临床试验中正在注入转基因 T 细胞以治疗白血病和淋巴瘤。该拨款申请验证了用于研究输注 T 细胞靶向肿瘤细胞能力的工具，并应用这种方法来提高我们对其在临床试验中的治疗益处的理解。这将使研究人员能够开发出产生更有效的 T 细胞的方法，以在下一代临床试验中靶向血液恶性肿瘤。