Modeling the complex genetics of congenital heart disease in mice

模拟小鼠先天性心脏病的复杂遗传学

基本信息

批准号：
9260066
负责人：
CECILIA W. LO
金额：
$ 76.85万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-15 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9260066
关键词：
Affect Anatomy Animal Model Autopsy Biological Assay CHD4 gene CHD7 gene CRISPR/Cas technology Cardiac Cardiovascular system Chemicals Cilia Clinical Clinical Research Clustered Regularly Interspaced Short Palindromic Repeats Complex Congenital Abnormality DNA Resequencing Defect DiGeorge Syndrome Echocardiography Ensure Ethylnitrosourea Exhibits Female Fetus Future Gene Targeting Genes Genetic Genetic Epistasis Genetic Predisposition to Disease Genetic Screening Genetic Variation Genome Genotype Heart Abnormalities Histopathology Human Human Genetics Hypoplastic Left Heart Syndrome Image Analysis Inbred Mouse Inbreeding Individual Induced Mutation Knock-in Live Birth MLL2 gene Mendelian disorder Modeling Mus Mutagenesis Mutant Strains Mice Mutation Mutation Analysis Nature Partner in relationship Pathogenesis Pathogenicity Patients Pediatric Cardiac Genomics Consortium Penetrance Phenotype Point Mutation Population Proteins Recovery Recurrence Risk Role Structural Congenital Anomalies Syndrome System Testing Transgenic Mice Ultrasonography Validation actionable mutation cardiovascular imaging ciliopathy congenital heart disorder design disease diagnosis disease phenotype disease-causing mutation exome sequencing experience fetal follow-up genetic analysis genetic approach genetic linkage analysis genetic pedigree genome editing imaging modality indexing male mouse genome mouse model mutant mutant mouse model novel offspring pediatric patients postnatal public health relevance reverse genetics

项目摘要

DESCRIPTION (provided by applicant) Congenital heart disease (CHD) is one of the most prevalent birth defects, affecting up to 1% of live births. While a genetic etiology is indicated by the finding of various syndromic forms of CHD, the genetic causes of CHD is still not well understood. As the genetic variability of the human population confounds human genetic analysis, we recently pursued a large scale ethylnitrosourea (ENU) mutagenesis screen in inbred C57BL6 mice to recover recessive mutations causing CHD. Phenotyping was conducted using noninvasive fetal echocardiography, an imaging modality used clinically for CHD diagnosis. From ultrasound scanning 100,000 fetuses, we recovered >250 CHD mutant mouse lines with a wide spectrum of CHD. We recovered 150 CHD causing mutations in 94 genes using whole exome sequencing analysis. Surprisingly, 50% of the CHD genes are cilia related, indicating a central role for cilia in CHD pathogenesis. Also unexpected was the recovery of many cilia related CHD genes encoding proteins known to physically interact, suggesting the hypothesis that CHD pathogenesis may occur via epistasis in the context of a CHD interactome network. This could contribute to the complex genetics associated with human CHD. To test this hypothesis, we plan to conduct a novel sensitized screen for the first systematic interrogation of the mouse genome for semi-dominant CHD mutations. The screen will be conducted using a driver mutation in Cep290, a cilia-CHD gene recovered in our recessive screen that is also clinically well described to exert genetic modifier effects, both in various human ciliopathies and in mutant mouse models. Our screen will entail ultrasound scanning G2 fetuses for CHD derived from females heterozygous for the Cep290 mutation mated to a G1 male heterozygous for ENU induced mutations. The resulting G2 offspring can only be double heterozygous for the Cep290 and ENU induced mutations (except when the ENU mutation is also in Cep290). Thus any CHD observed would reflect the effects of epistasis. Using this strategy, we expect to screen ~54,000 fetuses from 1800 G1 pedigrees, providing a systems approach to interrogate the role of cilia in the complex genetics of CHD not possible with a recessive screen. Mutation recovery will be carried out using whole mouse exome sequencing followed by genotyping analysis, and further linkage analysis conducted within and across multiple pedigrees. For validation of causal mutations identified in the screen, transgenic mouse models will be generated using CRISPR/Cas9 gene editing, followed by intercrosses with the Cep290 mutation to examine for evidence of epistasis. Overall, this novel sensitized screen will provide the first systematic interrogation of the mouse genome for semi-dominant mutations causing CHD. By using mutation in Cep290 as driver for this sensitized screen, we can further elucidate role of cilia in CHD pathogenesis. The mutations and animal models recovered in this sensitized screen will better model the complex genetics of human CHD and better inform future clinical studies interrogating the genetics etiology of CHD in the human population.

描述（由申请人提供）先天性心脏病（CHD）是Torth缺陷Y对各种综合症形式的CHD的发现之一，CHD的遗传原因仍未被很好地理解为人类遗传分析的遗传变异性，我们最近追求了大规模的乙基硝酸乙烯乙酸乙醇（ ENU在C57BL6小鼠中恢复了chd的突变在使用整个外显子组测序分析的94个基因中，有50％的CHD基因与CILIA相关，这表明CILIA在CHD发病机理中的中心作用可以可能能够使用与人类冠心病相关的一些复杂遗传学，以测试小鼠基因组的半主导chd突变的系统质量。在我们的屏幕上，源自cep290的男性杂合的男性杂合，源自cep290的男性杂合，源自cep290的男性杂合，也只能施加遗传的纤维病和突变的小鼠模型，它的基因筛选也很好地施加了遗传修饰。诱导的突变（除外，ENU突变也在CEP290中）。 CHD不可能使用凹进的筛选，然后是基因分析分析F在屏幕中的因果关系，将使用CRISPR/CAS9基因编辑，E CEP290突变产生转基因模型。小鼠基因组对半域突变的质疑是通过在CEP290中作为敏感筛查的驱动器的突变，我们可以进一步阐明纤毛在CHD发病机理中的突变和动物模型。人类冠心病的竞争更好的ORM未来临床研究询问了人口中冠心病的遗传学。