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Smad6 deficient zebrafish as a model for multifactorial craniosynostosis

Smad6缺陷斑马鱼作为多因素颅缝早闭的模型

基本信息

批准号：
10218617
负责人：
Shannon Fisher
金额：
$ 20.63万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10218617
关键词：
Adult Affect Alleles Animal Model Appearance BMP2 gene BMP7 gene Biological Biological Assay Biological Models Biological Process Cardiovascular system Categories Complex Conflict (Psychology)Congenital Abnormality Craniofacial Abnormalities Craniosynostosis Data Defect Development Disease Embryo Environmental Risk Factor Face Fibroblast Growth Factor Receptors Fishes Functional disorder Future Gene Dosage Gene Mutation Genes Genetic Genetic Engineering Genetic Models Genetic Risk Genetic study Head Heart Human Human Genetics Imaging Device In Situ Hybridization Incidence Individual Infant Injections Joint structure of suture of skull Limb structure Link MADH6 gene Missense Mutation Modeling Molecular Morphology Mus Mutant Strains Mice Mutation Pathogenesis Pathway interactions Patients Pattern Phenotype RNA Reporter Resources Risk Risk Factors Role Signal Transduction Skeletal system Surgical sutures System Testing Variant Zebrafish bone confocal imaging craniofacial cranium exome sequencing experimental study feasibility testing gastrulation gene function genetic risk factor genome wide association study histological stains human disease in vivo in vivo imaging inhibitor/antagonist insight mouse model mutant overexpression premature prenatal risk variant skeletal abnormality

项目摘要

Craniosynostosis (CS) is one of the most common craniofacial birth defects, affecting nearly 1/2000 infants. About 20% of cases are caused by mutations in single genes; most common are activating mutations in the genes encoding FGF receptors, with additional mutations identified in other genes, both transmitted and de novo. Existing mouse models for single gene CS have yielded significant insights into the molecular and developmental basis of CS and serves as a powerful illustration of the value of accurate animal models. However, the majority of CS cases are nonsyndromic, and the underlying genetic risk is more complex and probably also involves interactions with environmental factors. Recent human genetics studies have implicated several specific genes in some of these complex cases. Genome wide association studies (GWASs) of CS cases identified risk loci near the BMP2 and BMP7 genes. Subsequent whole exome sequencing (WES) of individual CS patients found a high incidence of mutations inactivating one copy of the gene encoding Smad6, a downstream inhibitor of BMP signaling. Individuals carrying the risk locus near BMP2 and one SMAD6 mutant allele reportedly had a greatly increased incidence of CS, suggesting a straightforward hypothesis that the combination of increased levels of BMP2 and reduced inhibitory Smad6 causes CS. Although this hypothesis has been challenged by more recent studies, there is also evidence that mutations in other genes can interact with SMAD6 to increase CS risk. To directly test these hypotheses, we propose to develop a zebrafish model to study the complex genetic risk factors underlying CS. We have generated inactivating mutations in the two zebrafish smad6 genes, and in Aim 1 we will characterize the phenotypes of fish lacking one or both copies of smad6a and smad6b. In Aim 2, we will take a two– pronged approach to assess genetic CS risk in zebrafish. First, we will cross the smad6 mutants with existing mutants for inhibitors of the BMP pathway, and with mutants for other genes implicated in human patients. We will assay the resulting fish for CS and other craniofacial and skeletal abnormalities, using histological staining and live confocal imaging. Second, we will use overexpression of human SMAD6 variants in zebrafish embryos as an efficient assay, to test the functional consequences of sequence variants identified in human patients. Through successful completion of our aims, we will establish an accurate model system for the complex genetics underlying the majority of CS cases. Zebrafish are amenable to in vivo imaging and direct manipulations during all stages of skull and suture formation, providing insight into the pathophysiology of CS. Also importantly, the model can be used to test other genes that may interact with smad6 mutations to increase CS risk, and will provide a sensitized genetic background to assess potentially contributing environmental factors.

颅缝闭塞（CS）是最常见的颅面出生缺陷之一，影响近1/2000