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

项目描述：生物学中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。细胞适应机制对维持健康的组织动态平衡至关重要，因为它们的失败破坏了组织的适应性，并导致了与年龄相关的疾病，如慢性炎症和癌症。人类的肠道里排列着上皮细胞，这些细胞在我们的身体和外部世界之间形成了一道物理屏障。这些细胞面临的一个关键挑战是如何保持这种屏障的完整性，以应对机械应力--当食物通过我们的肠道时，发生的生物物理信号，如拉伸、压缩和压力。近年来，机械力通过转录调节因子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激活上游的主要机械应力传感器仍然没有得到很好的表征。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(GPCRs)。我们最近发现了一种孤儿受体(目前未知的配体)，它偶联到微环境应激期间肠上皮细胞中YAP/TAZ的激活。然而，该受体的意义尚不清楚。令人兴奋的是，新获得的磷酸蛋白质组学数据表明，该受体向参与细胞-细胞连接、细胞外基质黏附和Rho GTP酶活性的蛋白质发出信号。由于这些通路被认为是紧密相连的，在上皮屏障功能和机械生物学中很重要，我们假设该受体是关键的机械传感器，控制着屏障对生物物理应激的完整性。在一个使用尖端技术的多学科研究项目中，如活细胞成像、3D有机物培养和2D肠道上皮机械传感模型，您将研究受体介导的信号转导如何在机械应激下塑造正常的肠道动态平衡和上皮屏障功能。将使用CRISPR-CAS9生成遗传功能丧失模型，该模型将与整合组学(RNAseq和蛋白质组学)相结合，以表征受体中介基因签名。将提供组学和生物信息学以及先进细胞生物学技术方面的培训，包括有机物培养、IncuCyte成像、共聚焦显微镜、RNAi和CRISPR-Cas9。你将在现代化的实验室中进行研究，并配备先进的边缘显微镜和蛋白质组学设施。了解这种受体在机械感觉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/或在包括炎症和癌症在内的许多疾病背景下促进组织再生。

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.