Cellular mechanotransduction - from the immune response to transcriptional regulation

细胞机械转导 - 从免疫反应到转录调节

• 批准号: 10693137
• 负责人: Arpita Upadhyaya
• 金额: $ 38.63万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10693137
• 关键词: Actins; Address; Adhesions; Binding; Biochemical; Biophysical Process; Cell Communication; Cell Nucleus; Cell physiology; Cells; Chromatin; Coupled; Cues; Cytoskeleton; Cytotoxic T-Lymphocytes; Data; Development; Disease; Disease Marker; Environment; Funding; Funding Mechanisms; Gene Expression; Gene Expression Regulation; Generations; Genetic Transcription; Genomics; Goals; Image; Image Analysis; Immune; Immune response; Immunotherapy; Intervention; Kinetics; Lymphocyte Activation; Measurement; Mechanics; Mediating; Methods; Microtubules; Molecular; Movement; National Institute of General Medical Sciences; Nature; Nuclear Hormone Receptors; Process; Receptor Signaling; Research; Resolution; Risk Factors; Signal Transduction; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Technology; Time; Tissues; Transcription Coactivator; Transcriptional Regulation; Visualization; adaptive immune response; cancer cell; cell killing; cytokine; density; design; diagnostic tool; flexibility; genome-wide; malignant breast neoplasm; mathematical model; mechanical force; mechanical properties; mechanical signal; mechanical stimulus; mechanotransduction; neoplastic cell; novel therapeutics; programs; quantitative imaging; response; single molecule; tool; transcription factor; tumor progression; wound healing

The overall goal of the research in my lab is to define the molecular mechanisms and functional consequences of cellular mechanotransduction – or how cells sense the mechanical properties of their microenvironment and launch appropriate functional responses. Cell-cell interactions, mediated by adhesion and signaling receptors, are highly dynamic and subject to cytoskeletal movements that impart substantial mechanical force at the interface. How cells combine mechanical and biochemical signals to carry out specific functions is not well understood. Our lab tackles this question in two contexts – the immune response in T cells and regulation of gene expression - using a combination of high (and super)-resolution imaging, force measurements, quantitative image analysis, genomics and mathematical modeling. As part of our NIGMS-funded research, we have recently demonstrated that T cell activation requires a close coordination of the actin and microtubule cytoskeletons in order to generate forces at the T cell receptor, which are transduced to biochemical signaling leading to T cell activation. We have shown that cytokine stimulation leads to modulation of cytoskeletal dynamics and force generation in cytotoxic T cells, facilitating the cytolytic response. We have also developed new methods for analysis of single molecule tracking data, which we have applied to study the dynamics and binding kinetics of transcription factors and relate them to genome-wide measurements. Over the next five years, we plan to continue to address the molecular mechanisms that mediate actin/microtubule crosstalk in T cells for the control of RhoA-mediated forces and how these cytoskeletal forces tune the mechanical coordination of cytotoxic T lymphocyte activation and their efficacy in killing cancer cells. Taking advantage of the flexible nature of the R35 funding mechanism, we will establish a new line of research that builds on our technological capabilities to examine how mechanical cues are relayed to the nucleus to regulate gene expression in a functionally appropriate manner and how mechanical cues interact with tissue-specific cues. We will use advanced tools for real-time visualization of nuclear hormone receptor and target gene transcription dynamics to interrogate 1) how substrate stiffness regulates chromatin accessibility and modulates the mobility of transcription factors and co-activators, with a particular focus on nuclear hormone receptors and 2) how biophysical mechanisms transduce changes in the mechanical environment into alterations in gene expression dynamics. Our research program will 1) elucidate how mechanical stimuli and biochemical signaling are coupled to orchestrate the adaptive immune response and 2) enable fundamental understanding of how mechanical properties of the microenvironment modulate gene expression, with implications for designing new targets for intervention in immune therapy and breast cancer.

我实验室研究的总体目标是定义分子机制和功能后果 细胞机理转导的转导 - 或细胞如何感知其微环境的机械性能和 启动适当的功能响应。通过粘合剂和信号接收器介导的细胞 - 细胞相互作用， 具有高度动态性并受到细胞骨架运动的影响，这些运动在 界面。细胞如何结合机械和生化信号来执行特定功能 理解齿。我们的实验室在两种情况下解决了这个问题 - T细胞中的免疫响应和调节 基因表达 - 使用高（和超级）分辨率成像的组合，力测量， 定量图像分析，基因组学和数学建模。作为我们NIGMS资助的研究的一部分，我们 最近证明，T细胞激活需要肌动蛋白和微管的密切配位 细胞骨架以在T细胞受体上产生力，然后将其翻译成生化信号传导 导致T细胞激活。我们已经表明，细胞因子刺激导致细胞骨架的调节 细胞毒性T细胞中的动力学和力产生，支持细胞溶解反应。我们也发展了 分析单分子跟踪数据的新方法，我们已应用于研究动力学和 结合转录因子的动力学，并将其与全基因组测量结果相关。在接下来的五个 几年，我们计划继续解决介导肌动蛋白/微管串扰T的分子机制 用于控制RhoA介导的力以及这些细胞骨架力如何调节机械的细胞 细胞毒性T淋巴细胞激活的协调及其在杀死癌细胞中的有效性。利用 R35资金机制的灵活性，我们将建立一个新的研究线，以我们的 研究如何将机械提示传递到核调节基因的技术能力 以功能合适的方式表达表达，以及机械线索与组织特异性提示的相互作用。 我们将使用先进的工具来实时可视化核马受体和靶基因 转录动力学询问1）底物刚度如何调节染色质的可及性和 调节转录因子和共激活因子的迁移率，特别关注核骑马 受体和2）生物物理机制如何在机械环境中转变为 基因表达动力学的改变。我们的研究计划将1）阐明机械刺激和 生化信号耦合以协调适应性免疫响应，2）启用基本 了解微环境的机械性能如何调节基因表达，并以 设计新靶标在免疫治疗和乳腺癌中的新目标的影响。