The Diversity Outbred Diabetes Project

多样性远交糖尿病项目

基本信息

批准号：
9763205
负责人：
Alan D Attie
金额：
$ 60.5万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9763205
关键词：
ATAC-seq Affect Animal Model B-Cell Development Beta Cell Bioinformatics Candidate Disease Gene Cell physiology Cellular biology Collaborations Communities Cyclic AMP-Dependent Protein Kinases Data Data Set Development Diabetes Mellitus Diet Embryo Epidemic Gene Deletion Generations Genes Genetic Genetic Screening Genetic Transcription Genetic Variation Genetic study Glucagon Human Human Genetics Inbred Strains Mice Insulin Insulin Resistance Islets of Langerhans Knock-out Knockout Mice Laboratories Lead Light Link Meiotic Recombination Molecular Mouse Strains Mus Non-Insulin-Dependent Diabetes Mellitus Nutrient Obesity Obesity Epidemic Pathway interactions Phenotype Physiological Play Population Population Genetics Predisposition Production Quantitative Trait Loci Regulation Research Research Design Resolution Risk Role Signal Transduction Single Nucleotide Polymorphism Stimulus Structure Structure of beta Cell of islet Testing The Jackson Laboratory Time Variant Vesicle Viral Work cell type detection of nutrient diabetes risk gene discovery gene function genetic resource genome wide association study human embryonic stem cell insight insulin secretion islet molecular phenotype novel novel therapeutic intervention patient stratification response risk variant trafficking transcription factor transcriptomics web site

项目摘要

PROJECT SUMMARY/ABSTRACT Type 2 diabetes (T2D) is the result of an increased demand for insulin along with an inability of pancreatic β- cells to meet this demand by secreting sufficient amounts of insulin. The majority of gene loci identified by human genetic studies of T2D affect β-cell development, mass, and/or function. Yet, there is a great deal that is unknown about how β-cells secrete insulin in response to nutrient stimuli. We carried out a genetic screen of insulin secretion in 233,447 pancreatic islets ex vivo that were isolated from 483 Diversity Outbred (DO) mice. The DO mice were derived from 8 inbred mouse strains representing ~80% of the genetic diversity of all mouse strains. They have as much genetic diversity as the entire human population; ~40 million single nucleotide polymorphisms. The DO mice have been maintained for >30 generations, accumulating enough meiotic recombinations to enable high-resolution mapping of pathophysiological phenotypes. Our genetic screen identified 30 gene loci that affect insulin secretion or content of insulin or glucagon. Using our pipeline for gene identification, we chose two genes for further study, Hunk and Zfp148. We derived a whole-body knockout of Hunk and a β-cell-specific knockout of Zfp148. Islets from both of these knockout mice secreted more insulin in response to nutrient stimulation than control mice. Hunk is a protein kinase, thus we will identify the substrates of Hunk to discover how it regulates insulin secretion. Zfp148 is a transcription factor. Thus, we will identify the direct transcriptional targets of Zfp148. We will apply our bioinformatic pipeline to identify additional genes from the 30 gene loci identified in our screen and provide them to the research community. Functional studies will be performed in genetically edited mice, genetically edited human ES-derived β-cells, and human islets. These studies will discover novel genes and pathways involved in β-cell function and T2D susceptibility.

项目概要/摘要 2 型糖尿病 (T2D) 是胰岛素需求增加以及胰腺 β- 功能障碍的结果细胞通过分泌足够量的胰岛素来满足这种需求。人类鉴定的大多数基因位点。 T2D 的遗传学研究会影响 β 细胞的发育、质量和/或功能，但仍有很多未知之处。关于β细胞如何分泌胰岛素以响应营养刺激我们进行了胰岛素的基因筛选。离体 233,447 个胰岛的分泌，这些胰岛是从 483 只多样性远交 (DO) 小鼠中分离出来的。小鼠源自 8 个近交系小鼠品系，代表了所有小鼠品系遗传多样性的约 80%。他们拥有与整个人类一样多的遗传多样性；约 4000 万个单核苷酸 DO小鼠已维持超过30代，积累了足够的减数分裂。重组以实现病理生理表型的高分辨率图谱。使用我们的基因管道确定了 30 个影响胰岛素分泌或胰岛素或胰高血糖素含量的基因位点。鉴定后，我们选择了两个基因进行进一步研究，Hunk 和 Zfp148，我们得到了 Hunk 和 Zfp148 的全身敲除。 Hunk 和 Zfp148 的 β 细胞特异性敲除小鼠的胰岛细胞分泌更多胰岛素。 Hunk 是一种蛋白激酶，因此我们将鉴定其底物。 Hunk 的研究发现 Zfp148 是一种转录因子，它如何调节胰岛素的分泌。我们将应用我们的生物信息管道来识别 Zfp148 的直接转录靶标。我们筛选中确定的 30 个基因位点并将其提供给研究界。在基因编辑小鼠、基因编辑人类 ES 衍生 β 细胞和人类胰岛中进行。研究将发现与 β 细胞功能和 T2D 易感性相关的新基因和途径。