Mechanisms of modifier effects on aganglionosis in the Sox10Dom model of HSCR

HSCR Sox10Dom 模型中修饰因子对神经节缺失的影响机制

• 批准号: 7874522
• 负责人: Lauren Walters
• 金额: $ 1.69万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7874522
• 关键词: Biological; Biological Assay; Biological Process; Candidate Disease Gene; Colon; Communities; Complex; Congenital Disorders; Congenital Megacolon; Development; Disease; Disease model; Distal; Embryo; Enteral; Enteric Nervous System; Etiology; Family member; Ganglia; Gastroenterology; Gastrointestinal tract structure; Gene-Modified; Generations; Genes; Genetic; Hereditary Disease; Immunohistochemistry; Individual; Knowledge; Modeling; Modification; Mus; Mutation; Myxoid cyst; Names; National Research Service Awards; Neural Crest; Pathogenesis; Patients; Penetrance; Phenotype; Play; Research; Role; Severities; Stem cells; Techniques; Transgenic Organisms; Variant; base; gene interaction; insight; migration; mouse model; mutant; nervous system development

DESCRIPTION (provided by applicant): Hirschsprung disease (HSCR) is a congenital disorder of the gastrointestinal tract that presents as a lack of ganglia in a variable portion of the distal colon due to deficits in the neural crest stem cells (NCSC) that give rise to the enteric nervous system. HSCR is a prime example of a complex genetic disease. Often HSCR cases are attributed to mutations in RET but mutations in other genes, including SOX10, can also cause aganglionosis. Approximately 50% of HSCR cases are of unknown etiology. Extent of aganglionosis varies widely among individuals and can even differ between family members. Studies in mouse models of HSCR demonstrate that modifier genes in the genetic background play a large role in determining both the penetrance and severity of aganglionosis. Understanding modifiers and gene-gene interactions that contribute to variation of HSCR will increase understanding of HSCR complexity and pathogenesis. Previous studies have established the utility of Sox10Dom mice in the study of HSCR. These mice recapitulate the aganglionosis seen in patients and have been used to identify modifiers of aganglionosis. The current study aims to demonstrate the interaction of a highly significant candidate gene with Sox10 to impact the aganglionic phenotype of the Sox10Dom HSCR model and will definite the biological effect of this interaction on enteric NCSC. Aim 1 will use BAG modification techniques and the generation of transgenic lines of mice to demonstrate that altered levels of the identified candidate gene modify the penetrance and severity of Sox10Dom aganglionosis. Aim 2 will use both immunohistochemistry and culture of purified NCSC to determine the effect of the modifying interaction on migration, proliferation, and developmental potential on NCSC. The results of this study will provide the gastroenterology community with a greater understanding of the causes of HCSR as well as the biological processes involved in the development of the enteric nervous system. Finally, characterization of the Sox10-modifier interaction will provide insight into how modifiers influence other complex diseases. This research will increase our understanding of the genetic basis of HSCR, help identify causes of aganglionosis in patients with non-classifiable disease, and increase knowledge of enteric nervous system development.

描述（由申请人提供）：巨结肠病（HSCR）是一种胃肠道先天性疾病，表现为远端结肠可变部分神经节缺失，原因是神经嵴干细胞（NCSC）缺陷导致肠神经系统发育。HSCR是一种复杂遗传疾病的典型例子。HSCR病例通常归因于RET突变，但包括SOX10在内的其他基因突变也可引起神经节病。大约50%的HSCR病例病因不明。神经节病的程度在个体之间差别很大，甚至在家庭成员之间也有差异。对小鼠HSCR模型的研究表明，遗传背景中的修饰基因在决定神经节病的外显率和严重程度方面起着重要作用。了解导致HSCR变异的修饰因子和基因-基因相互作用将增加对HSCR复杂性和发病机制的理解。先前的研究已经确立了Sox10Dom小鼠在HSCR研究中的实用性。这些小鼠再现了在患者中看到的神经节病，并已用于鉴定神经节病的修饰因子。目前的研究旨在证明一个高度显著的候选基因与Sox10的相互作用会影响Sox10Dom HSCR模型的神经节表型，并将确定这种相互作用对肠道NCSC的生物学效应。目的1将使用BAG修饰技术和转基因小鼠系的产生来证明所鉴定的候选基因水平的改变可以改变Sox10Dom神经节病的外显率和严重程度。目的2将使用免疫组织化学和纯化的NCSC培养来确定修饰相互作用对NCSC迁移、增殖和发育潜力的影响。这项研究的结果将为胃肠病学社区提供对HCSR的原因以及参与肠神经系统发育的生物学过程的更深入的了解。最后，sox10 -修饰子相互作用的表征将为修饰子如何影响其他复杂疾病提供见解。这项研究将增加我们对HSCR遗传基础的理解，有助于确定非分类疾病患者的神经节病的原因，并增加对肠神经系统发育的认识。