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Cardiac phenotyping of CHD candidate genes in Xenopus

非洲爪蟾 CHD 候选基因的心脏表型分析

基本信息

批准号：
8820278
负责人：
MARTINA BRUECKNER
金额：
$ 22.51万
依托单位：
YALE UNIVERSITY
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：
2014-03-10 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8820278
关键词：
Affect Alleles Anatomy Angiography Animal Model Animals Area Biological Assay Biological Models Biomechanics Candidate Disease Gene Cardiac Cardiac development Cardiology Cardiovascular system Childhood Clinical Color Congenital Abnormality Contrast Media Data Development Disease EFRAC Echocardiography Embryo Etiology Europe Future Genes Genetic Genetic screening method Genome Genomics Goals Grant Health Heart Hemoglobin Human Image Imaging Device Incidence Infant Infant Mortality Live Birth Methods Microscopic Modeling Monitor Newborn Infant Optics Patients Phase Phenotype Protocols documentation Rana Research Resolution Role Specificity Speed Stratification Stroke Volume Structure Tadpoles Technology Testing Validation Variant Xenopus abstracting base burden of illness cardiovascular visualization congenital heart disorder gene function high throughput screening human disease imaging modality imaging system knock-down novel optical imaging programs protein expression research study screening tomography tool

项目摘要

DESCRIPTION (provided by applicant): Congenital heart disease (CHD) occurs in approximately 8 out of 1000 live births and effects 1.3 million newborns per year worldwide. While there is evidence to indicate that CHD does have a genetic basis, most of CHD burden remains unexplained genetically. New genomics technologies can efficiently identify variations in the genomes of CHD patients, but only a small percentage have second unrelated alleles to validate them as disease causing. Therefore there is a pressing need to develop functional assays to evaluate these candidate genes for CHD. There are two main goals for these functional assays: 1) provide evidence supporting candidate genes as disease causing and 2) identify the mechanism for the candidate gene on normal development and the disease state. Here we develop Xenopus as a rapid model system for testing CHD genes and apply advanced optical imaging methods to detect cardiac phenotypes. Xenopus is as an important animal model of congenital heart disease: large numbers of embryos can be readily manipulated, protein expression can be knocked-down using antisense morpholino oligos, and the heart is easily visualized. To expand the CHD spectrum that can be modeling in Xenopus, we need better microscale cardiac imaging methods. During the R21 phase, we will test two technologies, optic coherence tomography (OCT) and our novel hemoglobin contrast subtraction angiography (HCSA) to demonstrate that microscale imaging of Xenopus can be used to screen CHD genomic hits. Optic coherence tomography is an optical imaging system that can capture microscopic structures at high acquisition speeds allowing high-resolution phenotyping of dynamic heart structures. Hemoglobin contrast subtraction angiography (HCSA) is a noninvasive, nondestructive, quantitative microangiographic method that exploits the hemoglobin as an endogenous flow contrast agent during color imaging enabling us to delineate abnormal structures as well as quantify biomechanical phenotypes. In the R33 phase, our overall goal is to apply these methods to facilitate detailed high-resolution structural phenotypin of tadpole hearts that can be used to quickly test CHD candidate genes for cardiac phenotypes. This will allow us to identify cardiac phenotypes in CHD candidate genes that have no previous role in cardiac development and serve as a springboard for future mechanistic studies.

描述（由申请人提供）：先天性心脏病（CHD）发生在大约8/1000活产婴儿和影响130万新生儿每年在世界各地。虽然有证据表明CHD确实有遗传基础，但大多数CHD负担仍然无法从遗传学上解释。新的基因组学技术可以有效地识别CHD患者基因组中的变异，但只有一小部分人具有第二个不相关的等位基因，以验证它们是否致病。因此，迫切需要开发功能检测来评估这些冠心病候选基因。这些功能测定有两个主要目标：1）提供支持候选基因作为致病基因的证据和2）鉴定候选基因对正常发育和疾病状态的机制。在这里，我们开发非洲爪蟾作为一个快速的模型系统，用于测试CHD基因，并应用先进的光学成像方法来检测心脏表型。非洲爪蟾是一种重要的先天性心脏病动物模型：大量的胚胎可以很容易地操作，蛋白质表达可以敲低使用反义吗啉代寡核苷酸，心脏很容易可视化。为了扩展可以在非洲爪蟾中建模的CHD谱，我们需要更好的微尺度心脏成像方法。在R21阶段，我们将测试两种技术，光学相干断层扫描（OCT）和我们的新型血红蛋白对比减影血管造影术（HCSA），以证明非洲爪蟾的微尺度成像可用于筛选CHD基因组命中。光学相干断层扫描是一种光学成像系统，可以以高采集速度捕获微观结构，从而允许对动态心脏结构进行高分辨率表型分析。血红蛋白对比减影血管造影术（HCSA）是一种非侵入性、非破坏性、定量微血管造影方法，其利用血红蛋白作为彩色成像期间的内源性血流造影剂，使我们能够描绘异常结构以及量化生物力学表型。在R33阶段，我们的总体目标是应用这些方法来促进蝌蚪心脏的详细高分辨率结构表型，可用于快速测试心脏表型的CHD候选基因。这将使我们能够识别CHD候选基因中的心脏表型，这些基因以前在心脏发育中没有作用，并为未来的机制研究提供跳板。