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: 10582540
• 负责人: Redwan Bhuiyan
• 金额: $ 4.32万
• 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/10582540
• 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; 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; Regulatory Element; Reporter; Research; Risk; Role; Scientist; Signal Transduction; Single Nucleotide Polymorphism; Stains; Stress; Techniques; Testing; Thapsigargin; The Jackson Laboratory; Time; Tissues; Training; Transcriptional Regulation; Tunicamycin; Untranslated RNA; Variant; Work; biological adaptation to stress; blood glucose regulation; career; cell type; cellular resilience; collaborative environment; doctoral student; endoplasmic reticulum stress; environmental stressor; epigenome editing; experience; functional genomics; genetic variant; genome editing; genome wide association study; genome-wide; in vivo; innovation; insulin secretion; islet; physiologic stressor; precision genomic medicine; programs; promoter; 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.

项目摘要 2型糖尿病（T2 D）是一种由遗传和环境因素引起的复杂疾病。全基因 关联研究（GWAS）已经在243个基因座（T2 D变体）上确定了403个关联信号， 遗传风险功能（表位）基因组分析强烈表明，非编码T2 D变体改变转录 T2 D基因在胰岛中的表达和靶基因的调节，但只有约20%的T2 D变异体引起胰岛中的变化。 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 因为内质网（ER）应激与胰岛功能障碍有关。然而，迄今为止的研究表明， 未评估T2 D变体是否或如何调节胰岛对ER应激的反应。我假设这些T2 D 变异体改变胰岛ER应激反应性CRE的使用或活性以及靶基因表达， 胰岛β细胞功能障碍或死亡。在我的初步数据分析中，我已经确定了ER应激反应 在胰岛中与407个T2 D变体重叠的克雷斯，并将这些克雷斯连接到22个推定的靶基因启动子 使用胰岛启动子捕获Hi-C图谱。在目标1中，我将全面评估T2 D变体对 使用染色质可及性数量性状基因座（caQTL）的ER应激响应性克雷斯的用途或活性 和大规模平行报告基因测定（MPRA）。在目标2中，我将确定假定的综合招聘考试- 人胰岛细胞中的靶向基因是其ER应激反应和存活所必需的， 使用CRISPR/Cas9表观基因组编辑平台进行表达。成功实现这些目标将产生 一组经功能表征的T2 D变体和一组经验证的下游“T2 D靶基因”， 调节胰岛功能和存活以响应中枢（病理性）生理应激。更广泛地说，我 我预计，我在这个机械的变体到功能项目中使用的原则和框架可能是 应用于表征T2 D变体在其他环境应激源和代谢的背景下的影响， 组织中重要的是，这个项目的完成将帮助我掌握当前的概念和国家的最先进的 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能： 广泛的机会来展示和发表我的研究。最后，我作为康州大学的一名博士生 健康和杰克逊基因组医学实验室不仅能让我在一个世界里接受指导- 类，高度协作的环境，但它将为我提供机会，继续磨练我的临床 在我的研究阶段期间和之后获得专业技能和专业经验。完成我的培训和发展 作为一名研究遗传和（表观）基因组的医生兼科学家，这项计划将是我未来职业生涯中至关重要的一步。 疾病相关变异的机制。