Genetic Analysis of Organ Patterning Defects in Ciliopathies

纤毛病器官模式缺陷的遗传分析

基本信息

批准号：
10477030
负责人：
ZHAOXIA SUN
金额：
$ 38.34万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10477030
关键词：
Affect Alleles Animal Model Animals Biology Birth CRISPR/Cas technology Cardiovascular system Cells Characteristics Chromosome 4 Cilia Complex Coupled Cyst Defect Development Diagnosis Diagnostic Disease Etiology Event Future Gait Gene Expression Profiling Genes Genetic Genetic Techniques Genome Health High-Throughput Nucleotide Sequencing Histologic Human Human Development Image Incidence Individual Infertility Kidney Length Light Link Live Birth Longitudinal Studies Lung Male Sterility Maps Mediating Mental Retardation Modeling Modification Mus Mutant Strains Mice Mutation Neonatal Nonsense Mutation Obesity Organ Organelles Organogenesis Other Genetics Pathology Pathway Analysis Patients Pattern Perinatal mortality demographics Phenocopy Phenotype Play Polydactyly Proteins Proteomics Regulation Role Set protein Severities Shapes Signal Pathway Signal Transduction Situs Inversus Stains Sterility Syndrome System Targeted Resequencing Testing Therapeutic Tissues VATER (vertebral defects-anal atresia-tracheoesophageal fistula-esophageal atresia-radial and renal dysplasia) association or syndrome base cell motility cell type ciliopathy cilium biogenesis craniofacial defined contribution experimental study genetic analysis genetic variant genomic locus human disease insight microCT microscopic imaging mutant novel skeletal trafficking vertebrate embryos

项目摘要

Abstract: Ciliopathies are an expanding group of human disorders whose cellular basis can be traced to defects in the formation or function of small, cellular organelles called cilia. Cilia are found on almost every cell type in the vertebrate body and thus ciliopathies can encompass defects in multiple organs and tissues. Strikingly, ciliopathies span a spectrum, ranging from infertility to neonatal lethality but the genetics underlying these divergent phenotypes are poorly understood. Defining the genes and genetic interactions that regulate cilia function is essential to understanding the etiology of these diseases as well as for developing future therapies. We have developed mice carrying a spontaneous mutation (hop) in the Ift56 gene into a new model for defining the genetic control of ciliopathy severity. Ift56 is a highly conserved cilia-localized protein that is required for cilia function in several model organisms. We recently discovered that while the Ift56hop phenotype is viable and mild on the Balb mouse background, it is lethal on the B6 background. The phenotype of the hop-B6 mutant models VACTERL Association in humans and overlaps with the Ift27 mutant ciliopathy phenotype. Our objectives are to 1) utilize the hop-B6 mutant as a means to understand VACTERL disorders, 2) define the function of Ift56 in mammalian cilia, and 3) isolate genetic loci that interact with the Ift56 mutation in order to elucidate the genetic landscape underlying ciliopathies. Our First Aim focuses on analyzing the tissue defects in Ift56-B6 mutants. Histological approaches coupled with gene expression analysis and microCT imaging will provide key insight into signaling and developmental patterning events controlled by Ift56 and the VACTERL-associated defects in hop-B6 mutants Aim2 explores the ciliary roles of Ift56, and how these are altered in the B6 and Balb backgrounds. Using candidate and proteomic approaches, we will determine the set of proteins that require Ift56 function for their localization and trafficking within cilia. Finally, we test the requirement of Ift56 for IFTB complex integrity. In our Third Aim we map, isolate and validate modifiers of the hop phenotype in the B6 and Balb backgrounds. We will also examine genetic interactions between Ift27 and hop mutants as well as the effects of the Balb modifier background on the Ift27 phenotype. These experiments will identify key genetic modifiers of ciliopathy severity and provide new targets for resequencing in ciliopathy patients. Together, the proposed studies will uncover the conserved functions of Ift56, a key cilia protein, by defining how Ift56 regulates cilia and organogenesis. More broadly, these studies have the opportunity to uncover unique insights into how genetic variants across the genome modify ciliopathy severity. Results from this study will shed key and novel insights into cilia biology and ciliopathies, and lay the groundwork for future diagnostic and therapeutic strategies.

抽象的：纤毛病是一组不断扩大的人类疾病，其细胞基础可以追溯到纤毛缺陷。称为纤毛的小型细胞器的形成或功能。纤毛几乎存在于所有细胞类型中脊椎动物体以及因此纤毛病可以涵盖多个器官和组织的缺陷。引人注目的是，纤毛病的范围很广，从不孕到新生儿致命，但这些疾病背后的遗传学人们对不同的表型知之甚少。定义调节纤毛的基因和遗传相互作用功能对于了解这些疾病的病因以及开发未来的治疗方法至关重要。我们已经将 Ift56 基因携带自发突变（hop）的小鼠培育成一种新模型定义纤毛病严重程度的遗传控制。 Ift56 是一种高度保守的纤毛定位蛋白，几种模式生物中纤毛功能所必需的。我们最近发现，虽然 Ift56hop 表型在 Balb 小鼠背景下是可行且温和的，在 B6 背景下是致命的。表型 hop-B6 突变体模型 VACTERL 与人类的关联以及与 Ift27 突变体纤毛病的重叠表型。我们的目标是 1) 利用 hop-B6 突变体作为了解 VACTERL 疾病的手段， 2) 定义 Ift56 在哺乳动物纤毛中的功能，以及 3) 分离与 Ift56 突变相互作用的遗传位点为了阐明纤毛病的遗传景观。我们的首要目标是分析 Ift56-B6 突变体的组织缺陷。组织学方法耦合基因表达分析和 microCT 成像将为信号传导和发育提供重要见解由 Ift56 控制的模式事件和 hop-B6 突变体中的 VACTERL 相关缺陷 Aim2 探讨了 Ift56 的纤毛作用，以及这些作用在 B6 和 Balb 背景中如何改变。使用候选和蛋白质组学方法，我们将确定需要 Ift56 功能的蛋白质组纤毛内的定位和运输。最后，我们测试了 Ift56 对 IFTB 复杂完整性的要求。在我们的第三个目标中，我们绘制、分离和验证 B6 和 Balb 中啤酒花表型的修饰因子背景。我们还将检查 Ift27 和啤酒花突变体之间的遗传相互作用及其影响 Ift27 表型上的 Balb 修饰符背景。这些实验将确定关键的遗传修饰剂纤毛病的严重程度并为纤毛病患者的重新测序提供新的目标。总之，拟议的研究将通过定义来揭示 Ift56（一种关键的纤毛蛋白）的保守功能 Ift56 如何调节纤毛和器官发生。更广泛地说，这些研究有机会揭示关于整个基因组的遗传变异如何改变纤毛病严重程度的独特见解。这项研究的结果将为纤毛生物学和纤毛病提供关键和新颖的见解，并为未来的诊断奠定基础和治疗策略。