Role of NRF2 in oxidative stress pathophysiology of diabetes kidney disease

NRF2 在糖尿病肾病氧化应激病理生理学中的作用

• 批准号: 2118207
• 金额: --
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2118207
• 关键词: Role; NRF2; oxidative; stress; pathophysiology

The prevalences of type 1 (T1D) and type 2 diabetes (T2D) are still increasing, but their aetiologies were until recently thought to be entirely distinct: islet beta-cell failure to produce sufficient insulin to compensate for insulin resistance in T2D versus autoimmune destruction of pancreatic islet beta cells in T1D. However, the discovery that a polymorphism of the GLIS3 gene predisposes to both T1D and T2D, through altering beta-cell sensitivity to stress, has highlighted beta-cell health as a common denominator.This project combines the unique strengths of the DIL (Todd) - in studying the genetic and molecular causes of T1D and its genetic overlaps with T2D - and the NNRCO (Johnson) -in studying the cell biology of pancreatic beta-cell survival and in implementing high-throughput screens and the latest imaging technology - to investigate the genetic and mechanistic bases underlying beta-cell health and diabetes.Our ongoing analyses show that >150 genes in over 60 T1D risk regions are expressed in beta cells and that several T1D-candidate variants colocalise to regulatory motifs active specifically in human islets and not in immune cells. However, stress conditions, similar to those encountered in the pre-diabetic pancreas, are likely to alter chromatin states and gene regulation dramatically.The proposed project has two parallel aims:-Investigate how genes in several newly identified T1D regions (some of which overlap with T2D risk loci) cause diabetes through beta-cell fragility. 2-Discover new genes/pathways required for beta-cell health by investigating the effects of stress and diabetes status on chromatin states and gene regulation in human donor islets.Aim 1 will use gene knockdown approaches to screen through protein-coding genes proximal to regulatory regions active specifically in islets. We will compare effects in beta cells (EndoC- betaH1 cells assembled into spheroids/pseudoislets and donor islets) subjected to chemical (thapsigargin), immune (cytokine treatment, co-culture with cytotoxic T cells) and genetic (e.g. GLIS3 loss-of-function) stresses versus non-stressed conditions.In aim 2 we will define the changes in chromatin states between stressed and non-stressed donor pancreatic islets to identify genes and regulatory regions that may otherwise appear to be quiescent in control islets, using ATAC-seq and RNA-seq. The recent miniaturisation of these methods will allow us to split the islets from one donor into several experimental groups. Integration of transcriptomics and proteomics data will enable the in silico assembly of the data into functional pathways. Depending on tissue availability, we aim to compare findings with those from diabetic versus non-diabetic donor islets.

1型糖尿病（T1 D）和2型糖尿病（T2 D）的患病率仍在增加，但它们的病因直到最近才被认为是完全不同的：胰岛β细胞无法产生足够的胰岛素来补偿T2 D中的胰岛素抵抗，而T1 D中胰岛β细胞的自身免疫性破坏。然而，发现GLIS 3基因的多态性通过改变β细胞对压力的敏感性而易患T1 D和T2 D，该项目结合了DIL（托德）和NNRCO（约翰逊）的独特优势-研究T1 D的遗传和分子原因及其与T2 D的遗传重叠-在研究胰腺β细胞存活的细胞生物学和实施高通量筛选和最新的成像技术，以调查β细胞健康和糖尿病的遗传和机制基础。我们正在进行的分析表明，超过60个T1 D风险区域中的>150个基因在β细胞中表达，并且一些T1 D-候选变体共定位于在人胰岛中而不是在免疫细胞中特异性活性的调节基序。然而，与糖尿病前期胰腺中遇到的情况类似，应激条件可能会显著改变染色质状态和基因调控。拟议的项目有两个平行的目标：-研究几个新发现的T1 D区域（其中一些与T2 D风险位点重叠）中的基因如何通过β细胞脆性导致糖尿病。2-通过研究压力和糖尿病状态对人类供体胰岛染色质状态和基因调控的影响，发现β细胞健康所需的新基因/途径。Aim 1将使用基因敲除方法筛选胰岛特异性活性调控区附近的蛋白质编码基因。我们将比较β细胞中（EndoC-β H1细胞组装成球状体/假胰岛和供体胰岛）进行化学处理，（毒胡萝卜素），免疫（细胞因子治疗，与细胞毒性T细胞共培养）和遗传在目的2中，我们将定义染色质状态在应激和非应激条件之间的变化。使用ATAC-seq和RNA-seq对应激供体胰岛进行鉴定，以鉴定在对照胰岛中可能以其他方式表现为静止的基因和调控区。这些方法的最新发展将使我们能够将来自一个供体的胰岛分成几个实验组。转录组学和蛋白质组学数据的整合将使数据的计算机组装成为功能途径。根据组织的可用性，我们的目标是比较结果与糖尿病与非糖尿病供体胰岛。