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

• 批准号: 8547802
• 负责人: Laurence J.N. Cooper
• 金额: $ 44.61万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 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/8547802
• 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; 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; screening; tool; tumor

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.

描述（由申请人提供）：免疫疗法注入抗体，疫苗和细胞是人类恶性肿瘤的有效治疗方法。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，可以实现有效的抗肿瘤作用。临床试验揭示了免疫疗法的优势，包括（i）定义的作用机理，（ii）定义的特异性和有害脱靶效应的降低，（iii）比常规方法低的毒性。基于对嵌合抗原受体（CAR）进行遗传修饰的T细胞的产物转移的产物细胞疗法（ACT），在治疗所有其他治疗方法的肿瘤难治的临床试验中表现出了巨大的希望。特别是，对CD19特异性的CAR+ T细胞的使用表明，CD19+慢性淋巴细胞性白血病（CLL）对常规疗法的难治性表现出显着的抗肿瘤作用。这些试验注入了多种基因修饰的T细胞的种群，这些细胞已传播到临床上足够的数字。虽然注入T细胞的治疗潜力取决于它们的持续性，但评估注入T细胞存活的免疫自动性研究目前仅限于（i）描述T-Cell接种物和（II）在输液后恢复的T细胞中T细胞的T细胞接种物和（II）全体群体的功能的功能。该赠款的应用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）对临床级CD19特异性CAR+ T细胞的效力，以适应临床级CD19特异性CAR+ T细胞的效力。我们的目标是使用高吞吐量单细胞纳米叶筛选（SNS），我们已经开发出来的，以进行深度定量功能表征（多重细胞因子分泌，表型，表型，细胞毒性，调节性T细胞的影响，TREG，TREG，TREG）的for pers+ T细胞的影响，并将这些数据与T细胞从患者中恢复为T细胞。这些数据将用于（i）量化注入的T细胞的功能以及抗肿瘤作用的潜力，以及（ii）在下一代临床试验中提高T细胞的产生，以提高功效。我们将在特定目标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谱。在特定的目标2中，我们将实施我们的方法，以量化收养的细胞的体内持久性。这将检验假设可以量化临床级T细胞的治疗潜力以及下一代临床试验的形成。