T Cell Receptor Forces: From Molecular Mapping to Cancer Therapeutic Triggering

T 细胞受体力：从分子图谱到癌症治疗触发

基本信息

批准号：
10247093
负责人：
Joshua Mark Brockman
金额：
$ 8.96万
依托单位：
HARVARD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10247093
关键词：
3-Dimensional Address Amino Acids Antibodies Antigens Atomic Force Microscopy Autoantigens Automobile Driving Binding Biochemical Biochemical Markers Blocking Antibodies CD28 gene CD3 Antigens CD8-Positive T-Lymphocytes Calcium Signaling Cell membrane Cell surface Cells Complex Cytotoxic T-Lymphocytes DNA DNA Binding Data Doctor of Philosophy ERBB2 gene Event Exhibits Flow Cytometry Fluorescence Fluorescence Polarization Image Imaging Device Imaging Techniques Immunologic Surveillance Immunosuppression Immunotherapeutic agent Immunotherapy In Vitro Intercellular Junctions Intravenous Lateral Ligands Light Link Major Histocompatibility Complex Malignant Neoplasms Maps Measurement Measures Mechanics Mediating Methods Modeling Molecular Molecular Probes Monoclonal Antibody HuM291 Mus Nanotechnology Nature Oligonucleotides Optics Peptide Fragments Peptide/MHC Complex Phage Display Polarization Microscopy Production Property Receptor Activation Research Research Project Grants Resolution Role Rupture Scanning Signal Transduction Single-Stranded DNA Solid Neoplasm Source Stains Surface Surface Antigens T-Cell Activation T-Cell Antigen Receptor Specificity T-Cell Receptor T-Lymphocyte Techniques Technology Testing Therapeutic Tissues Touch sensation Transgenic Mice Validation Work ZAP-70 Gene anti-PD-L1 therapy aptamer base cancer cell cancer immunotherapy cancer prevention cell killing chimeric antigen receptor T cells combat cytotoxicity dynamic system fluorophore follow-up immunoengineering immunogenic immunoregulation in vivo insight mechanical force nanoscale neoplastic cell novel optical traps overexpression prevent programmed cell death ligand 1 programmed cell death protein 1 receptor receptor binding recruit response single molecule small molecule temporal measurement tool treatment group tumor weapons

3-Dimensional Address Amino Acids Antibodies Antigens Atomic Force Microscopy Autoantigens Automobile Driving Binding Biochemical Biochemical Markers Blocking Antibodies CD28 gene CD3 Antigens CD8-Positive T-Lymphocytes Calcium Signaling Cell membrane Cell surface Cells Complex Cytotoxic T-Lymphocytes DNA DNA Binding Data Doctor of Philosophy ERBB2 gene Event Exhibits Flow Cytometry Fluorescence Fluorescence Polarization Image Imaging Device Imaging Techniques Immunologic Surveillance Immunosuppression Immunotherapeutic agent Immunotherapy In Vitro Intercellular Junctions Intravenous Lateral Ligands Light Link Major Histocompatibility Complex Malignant Neoplasms Maps Measurement Measures Mechanics Mediating Methods Modeling Molecular Molecular Probes Monoclonal Antibody HuM291 Mus Nanotechnology Nature Oligonucleotides Optics Peptide Fragments Peptide/MHC Complex Phage Display Polarization Microscopy Production Property Receptor Activation Research Research Project Grants Resolution Role Rupture Scanning Signal Transduction Single-Stranded DNA Solid Neoplasm Source Stains Surface Surface Antigens T-Cell Activation T-Cell Antigen Receptor Specificity T-Cell Receptor T-Lymphocyte Techniques Technology Testing Therapeutic Tissues Touch sensation Transgenic Mice Validation Work ZAP-70 Gene anti-PD-L1 therapy aptamer base cancer cell cancer immunotherapy cancer prevention cell killing chimeric antigen receptor T cells combat cytotoxicity dynamic system fluorophore follow-up immunoengineering immunogenic immunoregulation in vivo insight mechanical force nanoscale neoplastic cell novel optical traps overexpression prevent programmed cell death ligand 1 programmed cell death protein 1 receptor receptor binding recruit response single molecule small molecule temporal measurement tool treatment group tumor weapons

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项目摘要

T cells migrate through tissues, scanning cell surfaces and selectively destroying infected or malignant cells. The T cell interacts with other cells through the T cell receptor (TCR), recognizing antigen peptide fragment bound to the major histocompatibility complex (pMHC), on target cells. The TCR-pMHC bond is extremely specific, recognizing a single target pMHC among a myriad of other antigens. Despite the extreme importance of the TCR-pMHC bond to cancer prevention and immune surveillance, the origin of TCR specificity remains unclear. There is indirect evidence that TCR recognition of antigen utilizes uses mechanical force, specifically piconewton forces parallel the T cell membrane, to differentiate foreign antigen from self-antigen. Additionally, only one or two TCR-pMHC molecular bonds may be sufficient to trigger T cell activation, but tools to map pN mechanical events and to measure the orientation of these forces of do not exist, hindering progress in understanding T cell mechanobiology. My PhD research focuses on developing tools for molecular mechanobiology. I have invented a technique, Molecular Force Microscopy, capable of mapping the 3D orientation of piconewton molecular forces. I have also invented tension-PAINT, a super-resolved imaging technique capable of mapping single molecule cellular forces with ~10 nm spatial resolution. My F99 research has two focuses. First, I will apply Molecular Force Microscopy in conjunction with biochemical markers of T cell activation (e.g. Zap70-EGFP and calcium signaling) to test the hypothesis that TCR prefers forces parallel to the cell membrane to activate in response to antigen. Second, I will apply tension-PAINT to T cell forces to test the hypothesis that molecular forces recruit co-receptors in a force-dependent manner during T cell activation. This research will provide vital mechanistic details about force-mediated T cell activation. For my postdoctoral (K00) work, I will transition to developing immunotherapuetics. Immunotherapies, including engineered immune cells and PD-L1 blocking antibodies, have been deployed as anti-tumor therapies. However, immunotherapy is ineffective in many cancers. I hypothesize that TCR forces at the T cell-tumor junction can be leveraged to create new, force-activated immunotherapeutics. I will create a DNA-based container which I have termed the DNA origami antigen (DOA). The DOA will bind to tumor cells via cancer-specific antibodies. T cell forces will open the container, revealing a highly immunogenic payload that will stimulate cytotoxic T cell killing of cancer therapeutic. This research will result in a novel class of molecular force activated immunotherapeutics to combat cancer.

T细胞通过组织迁移，扫描细胞表面并有选择地破坏感染或恶性细胞。 T细胞通过T细胞受体（TCR）与其他细胞相互作用，识别与主要组织相容性复合物（PMHC）结合的抗原肽片段，靶细胞。 TCR-PMHC键非常具体，识别单个目标PMHC 在无数其他抗原中。尽管TCR-PMHC键对预防癌症和免疫监测，TCR特异性的起源尚不清楚。有间接的证据表明，TCR识别抗原利用使用机械力，特异性的piconewton力平行于T细胞膜，以区分外抗原与自我抗原。此外，只有一个或两个TCR-PMHC分子键可能足以触发T细胞激活，但要绘制PN机械事件并测量方向的工具这些不存在的力，阻碍了理解T细胞机械生物学的进展。我的博士研究重点是开发用于分子机械生物学的工具。我有发明了一种技术，即分子力显微镜，能够映射3D方向 Piconewton分子力。我还发明了张力粉末，这是一种超级分辨的成像能够用〜10 nm空间分辨率映射单分子细胞力的技术。我的 F99研究有两个重点。首先，我将在 T细胞激活的生化标记（例如ZAP70-EGFP和钙信号传导）测试假设TCR更喜欢平行于细胞膜的力来激活抗原。其次，我将向T细胞力施加张力粉，以测试分子的假设在T细胞激活期间，迫使募集的共受体。这项研究将提供有关力介导的T细胞激活的重要机理细节。对于我的博士后（K00）工作，我将过渡到开发免疫疗法。免疫疗法，包括工程的免疫细胞和PD-L1阻断抗体，具有被部署为抗肿瘤疗法。但是，免疫疗法在许多人中无效癌症。我假设可以利用T细胞脉冲连接处的TCR力来创建新的，力激活的免疫治疗药。我将创建一个基于DNA的容器称为DNA折纸抗原（DOA）。 DOA将通过癌症特异性与肿瘤细胞结合抗体。 T细胞力将打开容器，揭示高度免疫原性有效载荷会刺激癌症治疗的细胞毒性T细胞杀死。这项研究将导致小说分子力类型激活免疫疗法以对抗癌症。