Cellular mechanotransduction - from the immune response to transcriptional regulation

我实验室研究的总体目标是确定分子机制和功能后果 或细胞如何感知其微环境的机械特性和 启动适当的功能性反应。由黏附和信号受体介导的细胞间相互作用， 是高度动态的，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的功能还不是很好 明白了。我们的实验室在两个背景下解决了这个问题-T细胞的免疫反应和调节 基因表达-使用高(和超)分辨率成像、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 最近发现T细胞的激活需要肌动蛋白和微管的密切协调 为了在T细胞受体上产生作用力，细胞骨架被转导到生化信号 导致T细胞活化。我们已经证明，细胞因子刺激导致细胞骨架的调节 细胞毒性T细胞的动力学和作用力的产生，促进细胞溶解反应。我们还开发了 分析单分子跟踪数据的新方法，我们已经将其应用于研究动力学和 转录因子的结合动力学，并将它们与全基因组测量联系起来。在接下来的五年里 几年来，我们计划继续研究在T细胞中介导肌动蛋白/微管串扰的分子机制 用于控制RhoA介导力的细胞以及这些细胞骨架力如何调节机械 细胞毒T淋巴细胞活化的协调性及其对癌细胞的杀伤作用利用 R35资助机制的灵活性，我们将建立一个新的研究路线，建立在我们的 检查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特定组织的线索相互作用。 我们将使用先进的工具实时显示核激素受体和靶基因 转录动力学以询问1)底物硬度如何调节染色质的可及性和 调节转录因子和共激活子的流动性，特别是核激素 受体和2)生物物理机制如何将机械环境的变化转化为 基因表达动态的变化。我们的研究计划将1)阐明机械刺激和 生化信号被耦合来协调适应性免疫反应和2)使基本 了解微环境的机械特性如何调节基因表达， 为免疫治疗和乳腺癌干预设计新靶点的意义。

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.