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对各种环境应力的感觉，反应和适应。细胞适应的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的适应性，并导致与年龄相关的疾病，例如慢性炎症和癌症。人类的肠道衬有上皮细胞，它们在我们的身体和外界之间形成物理障碍。这些细胞的钥匙扣是如何维持对机械应力的障碍的完整性 - 随着食物的推动，诸如拉伸，压缩和压力等生物物理提示。在循环中，机械力通过下游co偶联器yap/taz的下游激活作为细胞行为的关键调节剂。然而，在这种情况下，YAP/TAZISTIVITION上游的机械应力的主要传感器仍然很差。细胞感知和响应其环境变化的一个重要方法是通过G蛋白偶联受体（GPCR）。我们最近确定了一个孤儿受体（当前未知的配体），该受体在微环境应激期间与YAP/TAZ活化伴随着YAP/TAZ激活。然而，这种受体感官尚不清楚。兴奋地，新获得的磷酸蛋白质组学数据表明，该受体信号是对参与细胞 - 细胞结合，细胞外基质粘附和Rho GTPase活性的蛋白质的。由于已知这些途径在上皮屏障功能和机械生物学中是密切联系的，因此我们假设该受体是巧妙的机械传感器，它可以控制屏障的完整性，以响应生物物理应力。在使用诸如活细胞成像之类的尖端技术的多学科研究计划中，肠上皮上皮的3D Organoid Cultureand 2D机械感应模型，您将研究受体介导的标志性正常肠道稳态和上皮屏障如何响应机械压力的响应。将使用CRISPR-CAS9生成遗传丧失功能障碍模型，该模型将与综合法（RNASEQ和prototomics）结合使用，以表征受体 - 介导基因特征。培训将以OMICS和BIOFORMATICS以及先进的细胞生物学技术提供，包括器官培养物，Incucyte Imaging，Compocalmicroscopicy，RNAi和CRISPR-CAS9。您将通过削减前卫和蛋白质组学设施支持的现代实验室进行研究。了解该受体在机械传感和屏障功能中的作用将为鉴定药物靶标铺平道路，从而可以防止在许多疾病情况下，包括炎症和癌症在内的许多疾病环境中促进健康组织稳态的细分/或促进组织再生。