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

项目简介：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的健康，并导致与年龄相关的疾病，如慢性炎症和癌症。人类肠道内衬上皮细胞，形成我们的身体和外部世界之间的物理屏障。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-生物物理线索，如拉伸，压缩和压力，发生在食物被推过我们的肠道。近年来，机械力通过下游转录辅助调节因子雅普/TAZ的激活成为细胞行为的关键调节因子。然而，在这种情况下，雅普/TAZ激活上游的机械应力的主要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种重要方式是通过G蛋白偶联受体（GPCR）。我们最近发现了一个孤儿受体（配体目前未知），耦合到雅普/TAZ激活肠上皮细胞在微环境应激。令人兴奋的是，新获得的磷酸化蛋白质组学数据表明，这种受体的信号蛋白参与细胞连接，细胞外基质粘附，Rho GT3活性。由于这些途径被认为是密切相关的，在上皮屏障功能和机械生物学的重要性，我们假设，这种受体是关键的机械传感器，控制屏障的完整性，响应生物物理应力。在一个多学科的研究计划中，使用尖端技术，如活细胞成像，3D类器官培养和肠上皮的2D mechanosensing模型，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，以应对机械应力。将使用CRISPR-Cas9生成遗传功能缺失模型，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，IncuCyte成像，共聚焦显微镜，RNAi和CRISPR-Cas9。您将在现代化的实验室中进行您的研究，这些实验室由切割边缘显微镜和蛋白质组学设施提供支持。了解这种受体在机械感知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.