Time-lapse Flow Cytometry for Kinetic Profiling of T-Cell Function

用于 T 细胞功能动力学分析的延时流式细胞术

• 批准号: 10699148
• 负责人: Sheldon J.J. Kwok
• 金额: $ 100.23万
• 依托单位: LASE INNOVATION INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-05 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10699148
• 关键词: Acceleration; Address; Adoptive Cell Transfers; Aliquot; Antibodies; Bar Codes; Biological Assay; Biological Markers; Cancer Patient; Cancer Vaccines; Cell Separation; Cell physiology; Cell surface; Cells; Chemicals; Chronic; Collaborations; Contractor; Data; Detection; Development; Enzyme-Linked Immunosorbent Assay; Exhibits; Flow Cytometry; Fostering; Goals; Hour; Human; Immune; Immunologics; Immunooncology; Immunotherapy; Individual; Interferon Type II; Interleukin-10; Interleukin-2; Kinetics; Lasers; Malignant Neoplasms; Marketing; Measurement; Measures; Methods; Monitor; Patients; Performance; Peripheral Blood Mononuclear Cell; Phase; Phenotype; Prediction of Response to Therapy; Process; Production; Protocols documentation; Quality Control; Reporting; Research; Research Personnel; Resistance; Resolution; Sampling; Scientist; Small Business Innovation Research Grant; Specificity; Standardization; Stimulus; T cell response; T-Lymphocyte; TNF gene; Technology; Time; Treatment Protocols; Treatment outcome; Validation; cancer cell; cancer immunotherapy; cellular imaging; commercialization; cytokine; exhaustion; immune checkpoint blockade; improved; in vitro Assay; individualized medicine; innovation; instrumentation; laboratory development; novel; particle; phenotypic biomarker; programmed cell death protein 1; research and development; response; systemic toxicity; vaccine development

Abstract Despite progress in immunotherapies, there are significant challenges to overcome to make them more broadly applicable to different cancers and more effective for patients. However, current efforts to improve adoptive cell therapies and immune checkpoint blockade as well as cancer vaccines are hampered by the lack of a method to characterize the functional and phenotypical changes of different subpopulations of T cells over time at high throughput. The goal of this SBIR Phase II project is to develop a novel flow-cytometry method that can solve this technological bottleneck. The time-lapse flow cytometry uses laser particle (LP) barcodes to track each cell across flow measurements taken at different time points. Built on demonstrated proof-of-concept data, this project is focused on establishing a set of novel assays for characterizing T cells in response to different stimuli in a time-resolved manner. The first specific aim is to demonstrate short-term time-lapse assays to profile cytokine secretion (t-SEC). Secretion of TNF, IFNγ and IL10 from millions of the same cells in response to different types of stimulation are measured at multiple timepoints over 24 hours. The second specific aim is to demonstrate long-term time-lapse assay to monitor phenotype (t-PHENO), including T-cell exhaustion over 10 days. The third specific aim is to validate the t-SEC and t-PHENO assays with patient samples. The time- resolved, high-throughput, high-parameter assays are expected to accelerate the development of more effective and durable immunotherapies for cancer (>$100 B global market). Beyond immunotherapy, time-lapse flow cytometry is also expected to be useful in fundamental immunological research and vaccine development.

抽象的 尽管免疫疗法取得了进展，但要克服的挑战仍有重大挑战 更广泛地适用于不同的癌症，对患者更有效。但是，当前 努力改善适应性细胞疗法和免疫检查点阻滞以及癌症 缺乏表征功能和表型的方法来阻碍疫苗 T细胞在高吞吐量下的不同亚群的变化。这个Sbir的目标 第二阶段项目是开发一种可以解决此技术的新型流程仪方法 瓶颈。延时流式细胞仪使用激光粒子（LP）条形码跟踪每个单元 在不同时间点进行的流量测量。建立在概念验证的基础上 数据，该项目的重点是建立一组新的测定法，以表征T细胞中的T细胞 以时间分辨的方式对不同刺激的反应。第一个具体目的是证明 短期延时阿萨斯剖面细胞因子分泌（T-SEC）。 TNF，IFNγ和 从不同类型的刺激响应不同类型的刺激的数百万个单元中的IL10在 多个时间点24小时。第二个具体目的是展示长期的延时 测定以监测表型（T-Pheno）的测定，包括10天内的T细胞耗尽。第三 具体目的是用患者样品验证T-SEC和T-Pheno分析。时间 - 已解决的高通量，高参数测定有望加速 癌症的更有效和耐用的免疫疗法（> $ 100 B全球市场）。超过 免疫疗法，延时流式细胞术也有用 免疫研究和疫苗开发。