SSA: Elucidation of epigenomic and transcriptional mechanisms controlling stress-induced Fkbp5 gene expression in the hippocampus

这个博士学位是一个令人兴奋的项目，从分子水平上深入了解压力是如何影响大脑的。压力是每个人生活的一部分。当压力环境给人带来极大的创伤时，当压力暴露成为慢性压力时，以及在存在潜在遗传脆弱性的情况下，它就会成为一个健康问题。最近，基因Fkbp5 （Klengel et al.）被描述为一种导致重度抑郁症和创伤后应激障碍的遗传易感性（所谓的单核苷酸多态性（SNP））。自然神经科学。2013)。该基因的蛋白产物Fkbp51在人和啮齿动物中分别降低了糖皮质激素受体结合应激激素皮质酮和糖皮质激素的敏感性。过度刺激糖皮质激素受体被认为对大脑有有害影响；因此，fkbp51在保持糖皮质激素受体活性在健康范围内发挥重要作用。因此，维持健康和幸福的Fkbp5基因的适当表达是非常重要的。然而，目前关于该基因是如何调控的信息有限。已知糖皮质激素受体刺激Fkbp5基因的表达，提高Fkbp51蛋白的细胞水平，从而导致受体对应激激素的敏感性降低。最近，我们发现体内应激后海马中Fkbp5基因中的糖皮质激素受体与所谓的糖皮质激素反应元件结合（Mifsud & Reul, Proc. Natl.Acad）。科学。美国2016年)。本项目的目的是阐明Fkbp5基因的活性如何受到糖皮质激素受体和表观遗传因素（如组蛋白修饰和DNA甲基化状态）的调节。您将应用一系列最先进的分子技术，包括染色质免疫沉淀（ChIP），定量PCR， hnRNA和mRNA分析和DNA甲基化分析，生物信息学分析，放射免疫分析以及包括手术在内的动物实验。为了更深入地了解体外Fkbp5基因特定片段的功能特性，您将使用前沿基因编辑Crispr/Cas9技术。该项目将由布里斯托尔大学的Johannes Reul教授和Oscar Cordero Llana博士以及埃克塞特大学的Jonathan Mill教授监督。

This PhD studentship is an exciting project to obtain insight into how stress impacts on the brain atthe molecular level. Stress is part of everybody's life. It can become a health problem when a stressfulsituation is extremely traumatic, when exposure to stress becomes chronic, and in cases where thereis an underlying genetic vulnerability. Recently, a genetic vulnerability (a so-called single nucleotidepolymorphism (SNP)) for developing major depression and posttraumatic stress disorder wasdescribed for the gene Fkbp5 (Klengel et al. Nature Neurosci. 2013). The protein product of this gene,Fkbp51, reduces the sensitivity of the glucocorticoid receptor for binding the stress hormones cortisolor corticosterone, glucocorticoid hormones in man and rodents, respectively. Excessive stimulation ofthe glucocorticoid receptor is thought to have detrimental effects on the brain; therefore, Fkbp51plays an important role in keeping glucocorticoid receptor activity within healthy limits. Accordingly,to maintain health and wellbeing appropriate expression of the Fkbp5 gene is of major importance.Presently, however there is only limited information available about how the gene is regulated.Glucocorticoid receptors are known to stimulate expression of the Fkbp5 gene and raise cellular levelsof Fkbp51 protein, thus resulting in reduction of receptor sensitivity to respond to stress hormone.Recently, we discovered that glucocorticoid receptors bind to so-called glucocorticoid-responseelements within the Fkbp5 gene in the hippocampus after stress in vivo (Mifsud & Reul, Proc. Natl.Acad. Sci. USA 2016).The aim of this project is to elucidate how the activity of the Fkbp5 gene is regulated by glucocorticoidreceptors and epigenetic factors like histone modifications and DNA methylation status in vivo. Youwill apply a range of state-of-the-art molecular techniques including chromatin immuno-precipitation(ChIP), quantitative PCR, hnRNA and mRNA analysis and DNA methylation analysis, bioinformaticsanalysis, radioimmunoassays as well as animal experimentation including surgeries. To obtain deeperinsight into the functional properties of particular segments of the Fkbp5 gene in vitro you will use thefront-line gene editing Crispr/Cas9 technology.This project will be supervised by Professor Johannes Reul and Dr Oscar Cordero Llana at the Universityof Bristol and Professor Jonathan Mill at the University of Exeter.