Dissecting the roles of type 2 diabetes-associated variants and effector genes in islet endoplasmic reticulum stress response

剖析 2 型糖尿病相关变异和效应基因在胰岛内质网应激反应中的作用

• 批准号: 10464542
• 负责人: Redwan Bhuiyan
• 金额: $ 4.23万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-15 至 2026-03-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10464542
• 关键词: 3-Dimensional; ANXA5 gene; ATAC-seq; Affect; Beta Cell; Binding; Biochemical; Biological Assay; Biological Process; Blood Glucose; CISH gene; CRISPR/Cas technology; Cell Death; Cell Nucleus; Cell Survival; Cell physiology; Cells; Cellular Stress; Cessation of life; Chemicals; Chromatin; Clinical Skills; Color; Communication; Complex; Coupled; DNA Sequence; Data; Data Analyses; Development Plans; Disease; Electrophoretic Mobility Shift Assay; Elements; Environmental Risk Factor; Etiology; European; Exhibits; Exposure to; Failure; Fellowship; Fluorescein-5-isothiocyanate; Fluorescence-Activated Cell Sorting; Functional disorder; Future; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Diseases; Genetic Risk; Genetic Transcription; Genetic study; Genome; Genomic medicine; Genomics; Goals; Health; Hi-C; Human; Insulin; Insulin Resistance; Islet Cell; Islets of Langerhans; Knowledge; Link; Maps; Measures; Mediating; Mentors; Meta-Analysis; Metabolic; Molecular; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Pancreas; Pathway interactions; Patients; Phase; Phenotype; Physicians; Physiology; Positioning Attribute; Process; Proteins; Publishing; Quantitative Trait Loci; Regulator Genes; Regulatory Element; Reporter; Research; Risk; Role; Scientist; Signal Transduction; Single Nucleotide Polymorphism; Stains; Stress; Techniques; Testing; Thapsigargin; The Jackson Laboratory; Three-dimensional analysis; Time; Tissues; Training; Transcriptional Regulation; Tunicamycin; Untranslated RNA; Variant; Work; base; biological adaptation to stress; blood glucose regulation; career; cell type; collaborative environment; doctoral student; endoplasmic reticulum stress; environmental stressor; epigenomics; experience; functional genomics; genetic variant; genome editing; genome wide association study; genome-wide; in vivo; innovation; islet; physiologic stressor; precision genomic medicine; programs; promoter; resilience; response; risk variant; sensor; skills; stress resilience; transcription factor; transcriptome sequencing

PROJECT SUMMARY Type 2 Diabetes (T2D) is a complex disease caused by both genetic and environmental factors. Genome-wide association studies (GWAS) have identified 403 association signals at 243 loci (T2D variants) that increase T2D genetic risk. Functional (epi)genomic analyses strongly suggest that non-coding T2D variants alter transcriptional regulation and target gene expression in pancreatic islets, but only ~20% of T2D variants elicit changes in islet cis-regulatory element (CRE) use or gene expression under steady state conditions. Environmental factors such as endoplasmic reticulum (ER) stress have been implicated in islet dysfunction. However, studies to date have not assessed if or how T2D variants modulate the response of islets to ER stress. I hypothesize that these T2D variants alter islet ER stress-responsive CRE use or activity and target gene expression to contribute to islet β cell dysfunction or death in T2D. In my preliminary data analysis, I have identified ER stress-responsive CREs that overlap 407 T2D variants in islets and connected these CREs to 22 putative target gene promoters using islet promoter capture Hi-C maps. In Aim 1, I will comprehensively assess the effects of T2D variants on the use or activity of ER stress-responsive CREs using chromatin accessibility quantitative trait locus (caQTL) and massively parallel reporter assays (MPRA), respectively. In Aim 2, I will determine if the putative CRE- targeted genes are required in human islet cells for their ER stress response and survival by altering their expression using CRISPR/Cas9 epigenomic editing platforms. Successful completion of these Aims will yield a functionally characterized set of T2D variants and a validated set of downstream 'T2D target genes’ that modulate islet function and survival in response to a central (patho)physiologic stressor. More broadly, I anticipate that the principles and framework I employ in this mechanistic variant-to-function project could be applied to characterize the effects of T2D variants in the context of other environmental stressors and metabolic tissues. Importantly, completion of this project will help me master current concepts and state-of-the-art techniques in genetics and functional genomics and increase my scientific communication skills through extensive opportunities to present and publish my studies. Finally, my position as an MD/PhD student at UConn Health and The Jackson Laboratory for Genomic Medicine will not only allow me to be mentored in a world- class, highly collaborative environment, but it will provide me with opportunities to continue honing my clinical skills and gain specialized experience during and after my research phase. Fulfilling my training and development plan will be a crucial step toward my future career as a physician-scientist studying the genetic and (epi)genomic mechanisms of disease-associated variants in patients.

项目概要 2 型糖尿病 (T2D) 是一种由遗传和环境因素引起的复杂疾病。 关联研究 (GWAS) 已在 243 个位点（T2D 变异）确定了 403 个关联信号，这些信号会增加 T2D 功能（表观）基因组分析强烈表明非编码 T2D 变异会改变转录。 胰岛中的调控和靶基因表达，但只有约 20% 的 T2D 变异会引起胰岛的变化 顺式调节元件（CRE）的使用或稳态条件下的基因表达等环境因素。 然而，迄今为止的研究表明，内质网（ER）应激与胰岛功能障碍有关。 没有评估 T2D 变异是否或如何调节胰岛对 ER 应激的反应，我敢于承认这些 T2D 变异。 改变变体胰岛 ER 应激反应性 CRE 的使用或活性以及靶基因表达，以促进 在我的初步数据分析中，我已经确定了 2 型糖尿病中的胰岛 β 细胞功能障碍或死亡。 CRE 与胰岛中的 407 个 T2D 变异重叠，并将这些 CRE 连接到 22 个假定的靶基因启动子 使用胰岛启动子捕获 Hi-C 图谱 在目标 1 中，我将全面评估 T2D 变异对的影响。 使用染色质可及性数量性状基因座 (caQTL) 的 ER 应激反应 CRE 的使用或活性 和大规模并行报告分析 (MPRA)，分别在目标 2 中，我将确定假定的 CRE-。 人类胰岛细胞需要靶向基因来通过改变其内质网应激反应和生存 使用 CRISPR/Cas9 表观基因组编辑平台的表达成功完成这些目标将产生一个 具有功能特征的一组 T2D 变异体和一组经过验证的下游“T2D 靶基因” 调节胰岛功能和存活以响应中枢（病理）生理应激源更广泛地说，我。 预计我在这个机械变体到功能项目中采用的原则和框架可以是 用于表征 T2D 变异在其他环境压力源和代谢背景下的影响 重要的是，完成这个项目将帮助我掌握当前的概念和最先进的技术。 遗传学和功能基因组学技术，并通过以下方式提高我的科学沟通技巧 最后，我作为康涅狄格大学医学博士/博士生的职位。 健康和杰克逊基因组医学实验室不仅能让我在一个世界中得到指导—— 班级，高度协作的环境，但它将为我提供继续磨练我的临床能力的机会 在我的研究阶段和研究阶段之后获得技能和专业经验。 计划将是我作为一名研究遗传和（表观）基因组的医师科学家未来职业生涯的关键一步 患者疾病相关变异的机制。