Functionalizing a T1D/T2D-associated locus within an intron of GLIS3.

功能化 GLIS3 内含子内的 T1D/T2D 相关位点。

• 批准号: 10750082
• 负责人: Lauren Clark
• 金额: $ 3.51万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-12-01 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750082
• 关键词: Affect; Automobile Driving; Back; Binding; Binding Sites; Cells; ChIP-seq; Chromatin; Chromosome 19; Chronic Disease; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; DNA; Data; Data Set; Diabetes Mellitus; Disease; Documentation; Electrophoretic Mobility Shift Assay; Enhancers; Exhibits; Fatty acid glycerol esters; Functional disorder; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Risk; Genetic Transcription; Genome; Globulins; Glucose; Goals; Healthcare; Healthcare Systems; Human; Hyperglycemia; Incubated; Individual; Institution; Introns; Knowledge; Label; Lead; Luciferases; Maps; Measures; Methods; Mus; NR5A2 gene; Non-Insulin-Dependent Diabetes Mellitus; Nuclear Extract; Oligonucleotides; POU2F1 gene; Patients; Phenotype; Positioning Attribute; Predisposition; Regulator Genes; Research; Role; Single Nucleotide Polymorphism; Sucrose; Testing; Tissues; Training; United States; Untranslated RNA; Variant; Wild Type Mouse; cost; diabetes risk; differential expression; experimental study; genetic approach; genome wide association study; genomic locus; improved; in vitro activity; in vivo; insulin secretion; interest; islet; male; mouse model; novel; novel strategies; pancreatic islet function; promoter; skills; tenure track; transcription factor; transcriptome sequencing; type I and type II diabetes; vector; western diet

PROJECT SUMMARY Diabetes mellitus has been characterized since antiquity, with the first documentation of this group of disorders dating back to 1500 BC1. While many advances have been made in the field in terms of treatment, there remains no cure for diabetes. This has become increasingly problematic for both the individuals and the entire healthcare system as rates of diabetes continue to rise. It is estimated that healthcare for individuals with diabetes costs over $200 billion per year in the United States alone2. Diminishing these costs and improving treatment options for patients are just a couple of the many reasons why diabetes-related research is so critical. I will use a genetic approach to more fully understand the initiation and progression of diabetes. In this application, I highlight a method that integrates human and mouse datasets to examine the role of intragenic and intronic regions of the genome in diabetes susceptibility. One such locus of interest resides in the first intron of the GLIS3 gene and has highly significant SNPs associated with both type 1 diabetes (T1D) and type 2 diabetes (T2D). To study this Glis3T1D/T2D locus, we determined the syntenic region in mouse and deleted 1729bp to create a novel mouse model (Glis31729 mouse). I found that these mice exhibit hyperglycemia, despite increased insulin secretion and glucose-evoked Ca2+ oscillations. My overall objective for this proposal is to functionalize this locus in an effort to elucidate its role in driving diabetes. My central hypothesis for the function of this locus is that it acts as an enhancer for Glis3 in islets, and that T1D/T2D-related SNPs within the locus alter its activity. I will test this hypothesis, and thus accomplish the goals of this proposal, by pursuing the following aims: 1) evaluate the enhancer activity of the Glis3T1D/T2D locus, 2) identify the influence of T1D/T2D-associated SNPs on transcription factor binding at the Glis3T1D/T2D locus, and 3) evaluate differentially expressed (DE) genes in the Glis3 1729 mice to reveal the role of the Glis3T1D/T2D locus on altering T1D and T2D risk. With the completion of these aims, I anticipate a more complete understanding of this non-coding locus, including how it regulates gene expression and affects both type 1 and type 2 diabetes susceptibility. Additionally, I believe this proposal will provide the necessary training for me to progress as an islet biologist and prepare me for a tenure-track position at a research institution.

项目摘要 糖尿病自古以来就有其特征，第一次记录这组糖尿病患者。 可以追溯到公元前1500年虽然在治疗方面已经取得了许多进展， 糖尿病仍然没有治愈的方法。这对个人和社区都变得越来越成问题。 随着糖尿病发病率的持续上升，整个医疗保健系统都受到了影响。据估计，个人的医疗保健 仅在美国，糖尿病每年花费超过2000亿美元2。降低这些成本， 糖尿病相关研究如此重要的原因有很多，其中之一就是患者的治疗选择。 我将使用遗传学方法来更全面地了解糖尿病的发生和发展。在这 应用程序，我强调了一种方法，整合人类和小鼠数据集，以检查基因内的作用， 糖尿病易感性基因组的内含子区域。一个这样的感兴趣的基因座位于第一内含子中， GLIS 3基因，具有与1型糖尿病（T1 D）和2型糖尿病相关的高度显著的SNP （T2D）。为了研究Glis 3 T1 D/T2 D基因座，我们在小鼠中测定了该基因的同线区，并删除了1729 bp， 一种新的小鼠模型（Glis 3 + 1729小鼠）。我发现这些老鼠表现出高血糖，尽管胰岛素增加 分泌和葡萄糖诱发的Ca 2+振荡。我这个提议的总体目标是使这个位点功能化 试图阐明它在糖尿病中的作用。我对这个基因座功能的主要假设是， 在胰岛中作为Glis 3的增强子，并且该基因座内的T1 D/T2 D相关SNP改变其活性。我将测试 这一假设，从而实现这一建议的目标，通过追求以下目标：1）评估 Glis 3 T1 D/T2 D基因座的增强子活性，2）鉴定T1 D/T2 D相关SNP对转录的影响 在Glis 3 T1 D/T2 D基因座的因子结合，和3）评估Glis 31729小鼠中的差异表达（DE）基因 揭示Glis 3 T1 D/T2 D基因座在改变T1 D和T2 D风险中的作用。随着这些目标的实现，我 预计对这个非编码基因座有更全面的了解，包括它如何调节基因表达 并影响1型和2型糖尿病的易感性。此外，我相信这项建议将提供 必要的培训，我进步作为一个胰岛生物学家，并准备在一个研究终身职位我 机构。