High-throughput Label-free Biosensor Platform for Rapid Detection of Antigen-specific T Cells with Single Cell Resolution

高通量无标记生物传感器平台，用于以单细胞分辨率快速检测抗原特异性 T 细胞

• 批准号: 10156407
• 负责人: Farshid Ghasemi
• 金额: $ 40万
• 依托单位: WEDDELL TECHNOLOGIES LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10156407
• 关键词: Address; Adoptive Cell Transfers; Algorithms; Alleles; Antibodies; Antigen-Presenting Cells; Antigens; Architecture; Biological Assay; Biosensor; Boston; Cancer Patient; Cell Adhesion; Cell Size; Cells; Cellular Assay; Chemistry; Clinical; Clinical Trials; Collaborations; Colorectal; Communicable Diseases; Cytotoxic T-Lymphocytes; Dana-Farber Cancer Institute; Data Analyses; Detection; Development; Diagnostic; Dose; Drug Combinations; Effectiveness; Elements; Engineering; Engravings; Equipment; Film; Flow Cytometry; Fluorescence Microscopy; Fluorochrome; Frequencies; Future; Glass; HLA Antigens; Health; Hematologic Neoplasms; Immobilization; Immunoassay; Immunotherapy; In Situ; Industry; Label; Laboratories; Lead; Liquid substance; Major Histocompatibility Complex; Malignant Neoplasms; Malignant neoplasm of ovary; Malignant neoplasm of pancreas; Massachusetts; Measures; Medical; Melanoma Cell; Metals; Methods; Microfluidics; Modeling; Monitor; Mutation; Needles; Non-Small-Cell Lung Carcinoma; Optical Instrument; Optics; Outcome; Output; Patient Monitoring; Patient-Focused Outcomes; Patients; Peptides; Peripheral Blood Mononuclear Cell; Personal Satisfaction; Phase; Population; Refractive Indices; Resistance; Resolution; Route; SILV gene; Semiconductors; Silicon; Slide; Small Business Innovation Research Grant; Specificity; Structure; Surface; Surface Plasmon Resonance; System; T-Cell Activation; T-Cell Immunologic Specificity; T-Cell Receptor; T-Lymphocyte; Technology; Testing; Thinness; Time; Tumor Antigens; Universities; antigen detection; antigen-specific T cells; cancer cell; cancer diagnosis; cancer therapy; chimeric antigen receptor T cells; commercialization; design; detection platform; digital; engineered T cells; fluorescence microscope; high throughput analysis; imaging system; immune checkpoint blockade; improved; innovation; manufacturing scale-up; medical schools; melanoma; neoplastic cell; novel; operation; peripheral blood; photonics; point of care; prognostic; prototype; rapid detection; refractory cancer; response; sensor; side effect; single cell analysis; standard of care; success; technology validation; tumor; tumor behavior; tumor immunology

Project Summary / Abstract Immunotherapy has become a rapidly growing segment of cancer treatment with impressive success across a spectrum of malignancies including melanoma, colorectal, and non-small cell lung cancers. Notwithstanding these advances, a significant fraction of patients fails to respond to immunotherapy and suffers from serious adverse side effects. While peptide-loaded major histocompatibility complex (pMHC) tetrameric structures allow the detection of antigen-specific T cells, a few antigen specificities can be detected in parallel because of limitations on the number of available fluorescent or metal labels. On the other hand, thousands of cells are needed for the workflow. Improved prognostic methods to monitor the specificity and functional behavior of tumor antigen-specific cytotoxic T cells is greatly needed to enhance the overall effectiveness of a range of immunotherapies, especially adoptive cell therapy (ACT), and provide better outcomes for cancer patients. This project develops a first-of-its-kind biosensor platform that enables the rapid and parallel detection of antigen- specific T cells. The proposed lab-on-chip technology allows detection and characterization at a single-cell level without requiring the use of labeling, complicated operational controls, or expensive equipment. As a result, the technology can be implemented in point-of-care settings and rapidly provide medical professionals with critical information, such as the ideal timing of future injected doses and any off-target effects. The key innovations behind the proposed technology include its high-throughput biosensor architecture, the ability to scale-up manufacturing using existing silicon foundries, label-free cell detection, simple operation and product design, and the implementation of novel algorithms of robust, real-time data analysis. Moreover, the commercialization of the proposed technology is facilitated by a mature semiconductor industry to achieve this high level of multiplexing in a small form factor. The proposed project focuses on engineering and optimization of the proposed biosensor platform and iterative development using six well-characterized tumor-antigens that are frequently recognized in melanoma patients. Peripheral blood mononuclear cells (PBMCs) from melanoma patients and from healthy donors will be used for analysis and technology validation. Successful completion of the project will provide a laboratory proof-of- concept, allowing the technology to move forward to a clinical setting where it can be used to monitor patients’ ongoing responses to immunotherapy, in specific checkpoint blockade and/or adoptive cell therapy. The tumor profiling market is projected to grow to about $12B by 2024, with the largest sector being immunoassays. If successful, the proposed technology will be a groundbreaking development in the cancer immunology toolbox, especially for early ex vivo identification of resistant tumor cell subpopulations, and help advance the effectiveness of cancer treatment for millions of people around the world.

项目摘要 /摘要 免疫疗法已成为癌症治疗的迅速增长，在 恶性肿瘤的光谱，包括黑色素瘤，结直肠癌和非小细胞肺癌。虽然 这些进展，很大一部分患者无法对免疫疗法做出反应，并且患有严重的患者 不良副作用。虽然负载肽的主要组织相容性复合物（PMHC）四聚体结构允许 检测抗原特异性T细胞，可以并行检测一些抗原规格 可用荧光或金属标签数量的限制。另一方面，成千上万的细胞是 工作流程所需。改进的预后方法可以监测肿瘤的特异性和功能行为 抗原特异性细胞毒性T细胞非常需要增强一系列范围的整体效率 免疫疗法，尤其是适应性细胞疗法（ACT），并为癌症患者提供更好的结局。 该项目开发了一个初始的生物传感器平台，可快速和平行地检测抗原 特定的T细胞。拟议的片上实验室技术允许在单细胞水平上检测和表征 不需要使用标签，复杂的操作控制或昂贵的设备。结果， 技术可以在护理点环境中实施，并迅速为医疗专业人员提供关键 信息，例如未来注入剂量的理想时机和任何脱靶效应。关键创新 拟议技术的背后包括其高通量生物传感器架构，扩展的能力 使用现有硅铸造厂，无标签的单元检测，简单操作和产品设计的制造， 以及实时数据分析的新型算法的实施。而且，商业化 拟议的技术是由成熟的半导体行业准备的，以实现这一高水平 以较小的形式多路复用。 拟议的项目着重于拟议的生物传感器平台的工程和优化和迭代 使用六种特征良好的肿瘤抗原的发育，这些肿瘤抗原经常在黑色素瘤患者中被识别。 来自黑色素瘤患者和健康供体的外周血单核细胞（PBMC）将用于 分析和技术验证。该项目的成功完成将提供实验室证明 概念，允许该技术继续前进到可用于监测患者的临床环境 在特定检查点阻滞和/或适应性细胞疗法中，对免疫疗法的持续反应。 到2024年，肿瘤分析市场预计将增长到约12B美元，最大的行业是 免疫测定。如果成功，拟议的技术将是癌症的开创性发展 免疫学工具箱，尤其是用于耐药性肿瘤细胞亚群的早期鉴定，并有助于 提高世界各地数百万人的癌症治疗的有效性。