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

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、反应和适应各种微环境压力。细胞适应机制对于维持健康组织的内稳态至关重要，因为它们的失败会破坏组织的适应性，并导致慢性炎症和癌症等与年龄相关的疾病。人类的肠道布满了上皮细胞，它们在我们的身体和外界之间形成了一道物理屏障。对于这些细胞来说，一个关键的挑战是如何在应对机械压力时保持屏障的完整性。机械压力是指食物被推入肠道时产生的拉伸、压缩和压力等生物物理信号。近年来，机械力已成为细胞行为的关键调节剂，通过下游激活转录共调节剂YAP/TAZ。然而，在这种情况下，YAP/ taz激活上游的主要机械应力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途径是通过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），它在微环境应激下与肠上皮细胞中的YAP/TAZ激活偶联。然而，这种感受器的感觉仍然是未知的。令人兴奋的是，新获得的磷酸化蛋白质组学数据表明，该受体信号涉及细胞-细胞连接，细胞外基质粘附和Rho GTPase活性的蛋白质。由于已知这些途径在上皮屏障功能和机械生物学中密切相关且重要，我们假设该受体是关键的机械传感器，在生物物理应激反应中控制屏障完整性。在一个多学科的研究项目中，使用尖端技术，如活细胞成像，3D类器官培养和肠上皮的2D机械传感模型，您将研究受体介导的信号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。遗传损失功能模型将使用CRISPR-Cas9生成，该模型将与整合组学（RNAseq和蛋白质组学）相结合，用于表征受体介导的基因特征。培训将提供组学和生物信息学以及先进的细胞生物学技术，包括类器官培养，诱导细胞成像，共聚焦显微镜，RNAi和CRISPR-Cas9。您将在先进的显微镜和蛋白质组学设备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，将为确定药物靶点铺平道路，这些靶点可以防止健康组织稳态的破坏和/或促进包括炎症和癌症在内的许多疾病背景下的组织再生。

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.