Enabling technologies to study how mechanics influence T cell function at the molecular and cellular levels

研究力学如何在分子和细胞水平上影响 T 细胞功能的技术

基本信息

批准号：
9438293
负责人：
Pui-Yan Victor Ma
金额：
$ 4.55万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-20 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9438293
关键词：
Adverse effects Affect Antigens Architecture Autoimmunity Binding Biochemical Biological Process Biotinylation Cancer Biology Cancer Control Cell Proliferation Cell Survival Cell physiology Cells Characteristics Chemicals Clinical Coculture Techniques Complex Coupled Cues Cytotoxic T-Lymphocytes DNA Data Doctor of Philosophy Enzyme-Linked Immunosorbent Assay Event Extracellular Matrix Flow Cytometry Genetic Transcription Goals Growth Heterogeneity Hydrogels Immune Immune Evasion Immune checkpoint inhibitor Immunosuppression Immunosuppressive Agents Immunotherapy Integrins Intercellular Junctions Label Ligation Malignant Neoplasms Maps Measures Mechanics Mediating Modeling Molecular Monitor Names Neoplasm Metastasis Outcomes Research PDCD1LG1 gene Patients Peptide/MHC Complex Phase Phosphorylation Play Postdoctoral Fellow Production Proteins Proteomics Receptor Activation Receptor Signaling Research Research Personnel Research Project Grants Role Series Signal Transduction Spectrum Analysis Stimulus Surface T cell response T-Cell Activation T-Cell Receptor T-Lymphocyte Technology Translating Up-Regulation Work Wound Healing antitumor effect base cancer cell cancer immunotherapy career cell behavior cell killing cytokine design immunological synapse innovation interdisciplinary approach killings mechanical force mechanical properties mimetics molecular scale nanosensors neoplastic cell oncology overexpression patient subsets physical property physical science protein E receptor function response scaffold single molecule success tissue regeneration training opportunity tumor tumor immunology tumor microenvironment tumor progression

项目摘要

Recent evidence has showed that the tumor microenvironment (TME) may form a sanctuary for immune suppression and evasion. Although Immunotherapies based on checkpoint inhibitors and chemical modulation of the TME have garnered success, many challenges remain, including the heterogeneity of patient response and the serious side effects resulting from systemic autoimmunity. These observations suggested that biochemical stimuli are not the only factor that suppresses T cell functions within the TME. One emerging concept in the field is that the physical properties of the TME such as extracellular matrix stiffness, composition, and architecture also contributes to cancer cell proliferation and survival. However, whether the mechanical properties of the TME specifically modulates T cell activity, and thus contributing to immune evasion, remains unclear. There are two overarching goals for this proposal. First, to better understand the role of mechanical forces in T cell receptor activation, and T cell functional responses. Second, to understand how the physical aspects of TME affect T cell/tumor interactions and T cell function. My PhD work is focused on developing enabling technologies to study mechanobiology at the molecular scale, with a particular focus on the roles of mechanical forces in T cell activation. My work has shown that i) the T cell receptor transmits pN forces to its antigen during initial recognition, and in immunological synapse and ii) T cells use mechanical energy to discriminate antigens during the earliest step of T cell activation. For my remaining F99 phase, I will focus on investigating whether mechanical forces are important for T cell function. To achieve this goal, I will use recently developed proximity labeling technologies to identify mechanosensitive proteins that mediate T cell signaling. This strategy will allow for using proteomic analysis to determine the mechanical interactome (mechanome) in T cells. Then, I will determine whether mechanical forces affect long term T cell biological functions using ELISA and flow cytometry coupled with RNA-SEQ. Overall, the results from this integrative -omics study will offer better understanding of how T cells use mechanical energy to potentiate their biological functions. For my postdoc studies (K00 phase), I aim to understand how the physical aspects of TME affect cytotoxic T cell function (cancer killing). The goal is to quantify how ECM mechanics alter T cell function. This data will support the hypothesis that TME contributes to metastasis by enhancing immune evasion through physical mechanisms. The significance of this work pertains to developing new strategies for promoting T cell anti-tumor response. I propose to work in a lab that employs 3D matrices that mimic the TME and to use these scaffolds for co-culture of T cells and tumor grafts. The work will better define the physical parameters of the matrix (e.g. the extent of hydrogel stiffness, physical cues composition, and architecture) that affect T cell/tumor interaction and T cell function. Collectively, the work from both F99 and K00 phases will provide new fundamental understanding on the physical basis of T cell functions at molecular level and cellular levels. The outcome of this research may offer new design principles for targeted cancer immunotherapies.

最近的证据表明，肿瘤微环境（TME）可能形成免疫抑制的庇护所和逃避。尽管基于检查点抑制剂的免疫疗法和TME的化学调制具有获得成功，仍然存在许多挑战，包括患者反应的异质性和严重的一面全身自身免疫引起的影响。这些观察结果表明，生化刺激不是只有抑制TME内T细胞功能的因素。该领域的一个新兴概念是物理 TME的特性，例如细胞外基质刚度，组成和结构也有助于癌症细胞增殖和生存。但是，TME的机械性能是否特异性调节T细胞活性，从而导致免疫逃避，尚不清楚。该提案有两个总体目标。首先，更好地了解机械力在T细胞受体激活和T细胞功能反应中的作用。其次，了解TME的物理方面如何影响T细胞/肿瘤相互作用和T细胞功能。我的博士学位工作的重点是开发能够在分子上研究机械生物学的能力尺度，特别关注机械力在T细胞激活中的作用。我的工作表明我）细胞受体在初始识别过程中将PN力传递到其抗原，在免疫突触中，II）T细胞在T细胞激活的最早步骤中，使用机械能区分抗原。对于我剩下的F99 阶段，我将专注于研究机械力是否对T细胞功能很重要。为了实现这一目标，我将使用最近开发的接近标记技术来识别介导T的机械敏感蛋白细胞信号传导。该策略将允许使用蛋白质组学分析来确定机械相互作用组（机制）在T细胞中。然后，我将确定机械力是否影响长期T细胞生物学功能使用ELISA和流式细胞仪与RNA-Seq结合。总体而言，这项综合词组研究的结果将可以更好地了解T细胞如何使用机械能来增强其生物学功能。对于我的博士后研究（K00阶段），我旨在了解TME的物理方面如何影响细胞毒性 T细胞功能（癌症）。目标是量化ECM力学如何改变T细胞功能。这些数据将支持 TME通过通过物理机制增强免疫逃避的假设有助于转移。这项工作的意义与制定促进T细胞抗肿瘤反应的新策略有关。我建议在使用模仿TME的3D矩阵的实验室工作，并将这些脚手架用于共同文化 T细胞和肿瘤移植物。该工作将更好地定义矩阵的物理参数（例如水凝胶的程度影响T细胞/肿瘤相互作用和T细胞功能的刚度，物理提示组成和结构）。总体而言，F99和K00阶段的工作将为身体提供新的基本了解 T细胞功能在分子水平和细胞水平上的基础。这项研究的结果可能会提供新的设计靶向癌症免疫疗法原理。