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

• 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
• 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

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细胞在组织中迁移，扫描细胞表面并选择性地破坏感染的细胞， 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体（pMHC）结合的抗原肽片段， 靶细胞TCR-pMHC键是非常特异的，识别单个靶pMHC 在无数其他抗原中。尽管TCR-pMHC结合对TCR-pMHC结合的极端重要性， 尽管TCR在癌症预防和免疫监视中发挥重要作用，但TCR特异性的起源仍不清楚。 有间接证据表明TCR识别抗原利用机械力， 特别是皮牛顿力平行T细胞膜，以区分外来抗原， 自体抗原另外，仅一个或两个TCR-pMHC分子键可能足以 触发T细胞激活，但工具映射pN机械事件和测量方向 这些力量的不存在，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分子机械生物学的工具。我有 发明了一种技术，分子力显微镜，能够映射的3D方向， 皮牛顿分子力我还发明了张力涂料，一种超分辨率成像技术， 能够以~10 nm的空间分辨率绘制单分子细胞力的技术。我 F99研究有两个重点。首先，我将应用分子力显微镜结合 T细胞活化的生物化学标志物（例如Zap 70-EGFP和钙信号传导）来测试T细胞活化。 假设TCR偏好平行于细胞膜的力来响应于 抗原的其次，我将应用张力涂料T细胞的力量，以测试假设，分子 力在T细胞活化期间以力依赖性方式募集共受体。本研究 将提供关于力介导的T细胞活化的重要机制细节。 对于我的博士后（K 00）工作，我将过渡到开发免疫疗法。 免疫疗法，包括工程免疫细胞和PD-L1阻断抗体， 被用于抗肿瘤治疗。然而，免疫疗法在许多情况下是无效的。 癌的我假设T细胞-肿瘤连接处的TCR力量可以被利用来创造 新的原力激活免疫疗法我将创建一个基于DNA的容器， 称为DNA折纸抗原（DOA）。DOA将通过癌症特异性结合至肿瘤细胞。 抗体的T细胞的力量会打开容器，露出一个高度免疫原性的有效载荷， 将刺激细胞毒性T细胞杀死的癌症治疗剂。这项研究将产生一部小说 分子力激活的免疫疗法来对抗癌症。