Understanding receptor-mediated mechanosensing and signalling in cell barrier function during tissue homeostasis and stress responses

了解组织稳态和应激反应期间细胞屏障功能中受体介导的机械传感和信号传导

• 批准号: 2888176
• 金额: --
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2888176
• 关键词: Understanding; receptor; mediated; mechanosensing; signalling

Project description:A fundamental challenge in biology is understanding how our cells sense, respond, and adapt to a variety ofmicroenvironmental stresses. Mechanisms of cellular adaptation are crucial for maintaining healthy tissue homeostasis,as their failure undermines tissue fitness and contributes to age-related diseases such as chronic inflammation andcancer. The human gut is lined with epithelial cells that form a physical barrier between our bodies and the outside world. A keychallenge for these cells is how to maintain the integrity of this barrier in response to mechanical stress - thebiophysical cues such as stretch, compression and pressure that occur as food is pushed through our gut. In recentyears, mechanical forces have emerged as key regulators of cell behaviour through downstream activation of thetranscriptional co-regulators YAP/TAZ. However, the primary sensors of mechanical stresses upstream of YAP/TAZactivation in this context remain poorly characterised. An important way that cells sense and respond to changes in their environment is through G protein-coupled receptors(GPCRs). We recently identified an orphan receptor (ligands currently unknown) that couples to YAP/TAZ activation inintestinal epithelial cells during microenvironmental stress. However, what this receptor senses remains unknown.Excitingly, newly acquired phosphoproteomics data suggest this receptor signals to proteins involved in cell-celljunctions, extracellular matrix adhesion, and Rho GTPase activity. Since these pathways are known to be closelyinterlinked and important in epithelial barrier function and mechanobiology, we hypothesise that this receptor is acritical mechanosensor that controls barrier integrity in response to biophysical stress. In a multidisciplinary research programme using cutting-edge techniques such as live-cell imaging, 3D organoid cultureand 2D mechanosensing models of the intestinal epithelium, you will investigate how receptor-mediated signallingshapes normal intestinal homeostasis and epithelial barrier function in response to mechanical stress. Genetic loss offunction models will be generated using CRISPR-Cas9, which will be combined with integrative omics (RNAseq andproteomics) for characterisation of receptor-mediate gene signatures. Training will be provided in omics andbioinformatics as well as advanced cell biology techniques including organoid culture, IncuCyte imaging, confocalmicroscopy, RNAi and CRISPR-Cas9. You will carry out your research in modern laboratories supported by cutting edgemicroscopy and proteomics facilities. Understanding the role of this receptor in mechanosensing and barrier functionwill pave the way for the identification of drug targets that could prevent the breakdown of healthy tissue homeostasisand/or promote tissue regeneration in a number of disease contexts including inflammation and cancer.

项目描述：生物学的一个基本挑战是了解我们的细胞如何感知、响应和适应各种微环境压力。细胞适应机制对于维持健康的组织稳态至关重要，因为它们的失败会破坏组织健康并导致慢性炎症和癌症等与年龄相关的疾病。人类肠道内衬有上皮细胞，形成我们的身体和外界之间的物理屏障。这些细胞面临的一个关键挑战是如何保持这一屏障的完整性，以应对机械应力——食物被推入肠道时产生的生物物理线索，如拉伸、压缩和压力。近年来，机械力通过转录共调节因子 YAP/TAZ 的下游激活而成为细胞行为的关键调节因子。然而，在这种情况下，YAP/TAZ 激活上游机械应力的主要传感器的特征仍然很差。细胞感知和响应环境变化的一个重要方式是通过 G 蛋白偶联受体 (GPCR)。我们最近发现了一种孤儿受体（目前未知的配体），它在微环境应激期间与肠上皮细胞中的 YAP/TAZ 激活偶联。然而，这种受体的感觉仍然未知。令人兴奋的是，新获得的磷酸化蛋白质组学数据表明，这种受体向参与细胞-细胞连接、细胞外基质粘附和 Rho GTP 酶活性的蛋白质发出信号。由于已知这些途径在上皮屏障功能和机械生物学中紧密相连且重要，因此我们假设该受体是控制屏障完整性以响应生物物理应激的关键机械传感器。在使用活细胞成像、3D 类器官培养和肠上皮 2D 机械传感模型等尖端技术的多学科研究项目中，您将研究受体介导的信号传导如何塑造正常的肠道稳态和上皮屏障功能以响应机械应力。将使用 CRISPR-Cas9 生成遗传功能丧失模型，该模型将与综合组学（RNAseq 和蛋白质组学）相结合，用于表征受体介导的基因特征。将提供组学和生物信息学以及先进细胞生物学技术的培训，包括类器官培养、IncuCyte 成像、共焦显微镜、RNAi 和 CRISPR-Cas9。您将在由尖端显微镜和蛋白质组学设施支持的现代实验室中进行研究。了解这种受体在机械传感和屏障功能中的作用将为识别药物靶点铺平道路，这些药物靶点可以防止健康组织稳态的破坏和/或促进炎症和癌症等多种疾病背景下的组织再生。