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: 10406710
• 负责人: Arpita Upadhyaya
• 金额: $ 23.18万
• 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/10406710
• 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; Five-Year Plans; Funding; Funding Mechanisms; Gene Expression; Gene Expression Regulation; Generations; Genetic Transcription; Genomics; Goals; 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; Risk Factors; Signal Transduction; T cell regulation; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Time; Tissues; Transcriptional Regulation; Visualization; adaptive immune response; cancer cell; cell killing; cytokine; density; design; diagnostic tool; flexibility; genome-wide; high resolution imaging; 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 细胞机械转导的研究——或者细胞如何感知其微环境的机械特性以及 启动适当的功能反应，由粘附和信号受体介导， 具有高度动态性，并受到细胞骨架运动的影响，从而在细胞骨架上施加大量机械力 细胞如何结合机械信号和生化信号来执行特定功能尚不清楚。 我们的实验室在两个方面解决了这个问题——T 细胞的免疫反应和 T 细胞的调节。 基因表达——结合使用高分辨率（和超）分辨率成像、力测量、 作为 NIGMS 资助研究的一部分，我们进行了定量图像分析、基因组学和数学建模。 最近证明 T 细胞激活需要肌动蛋白和微管的密切协调 细胞骨架，以便在 T 细胞受体上产生力，该力被转导为生化信号传导 我们已经证明细胞因子刺激会导致细胞骨架的调节。 我们还开发了细胞毒性 T 细胞的动力学和力生成，促进细胞溶解反应。 分析单分子追踪数据的新方法，我们已将其应用于研究动力学和 转录因子的结合动力学并将其与接下来的五年内的全基因组测量联系起来。 多年来，我们计划继续解决 T 中介导肌动蛋白/微管串扰的分子机制 细胞控制 RhoA 介导的力以及这些细胞骨架力如何调节机械力 细胞毒性 T 淋巴细胞激活的协调及其杀死癌细胞的功效。 鉴于 R35 资助机制的灵活性，我们将在我们的基础上建立一条新的研究路线 研究机械信号如何传递到细胞核以调节基因的技术能力 以功能适当的方式表达以及机械线索如何与组织特异性线索相互作用。 我们将使用先进的工具对核激素受体和靶基因进行实时可视化 转录动力学询问 1) 底物硬度如何调节染色质可及性和 调节转录因子和共激活因子的流动性，特别关注核激素 受体和 2) 生物物理机制如何将机械环境的变化转化为 我们的研究计划将 1) 阐明机械刺激如何改变 生化信号耦合以协调适应性免疫反应，并且 2) 实现基本的 了解微环境的机械特性如何调节基因表达， 设计干预免疫治疗和乳腺癌新靶点的意义。