Genetic analysis of the ciliopathies ARPKD and Meckel syndrome

ARPKD 和梅克尔综合征纤毛病的遗传分析

• 批准号: 8605533
• 负责人: Peter C. Harris
• 金额: $ 30.81万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-15 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8605533
• 关键词: Alleles; Animal Model; Animals; Autosomal Dominant Polycystic Kidney; Autosomal Recessive Polycystic Kidney; Biliary; Caenorhabditis elegans; Categories; Cell model; Cessation of life; Cilia; Complex; Cyst; DNA; Data; Defect; Development; Diagnostic; Digit structure; Disease; Etiology; Exons; Eye; Family; Family Study; Family member; Fibrosis; Genes; Genetic; Genetic Epistasis; Genetic Heterogeneity; Genetic Load; Goals; Grant; Hepatic Cyst; Hereditary Disease; Heterogeneity; Inborn Genetic Diseases; Individual; Inherited; Kidney; Kidney Failure; Knock-in Mouse; Link; Liver; Liver Fibrosis; Methods; Morbidity - disease rate; Mutation; Neonatal; Nephronophthisis; Neuraxis; Notoencephalocele; Oligogenic Traits; Organ; PKD1 gene; Pathogenicity; Patients; Phenotype; Play; Polycystic Kidney Diseases; Polydactyly; Population; Population Study; Protein Family; Proteins; Publications; Publishing; Role; Surface; Syndrome; System; Testing; Textbooks; Transcript; Variant; Zebrafish; ciliopathy; digital; disease phenotype; disease-causing mutation; exome; genetic analysis; in utero; kinetosome; mouse model; next generation sequencing; paralogous gene; prognostic; public health relevance; tool

DESCRIPTION (provided by applicant): In the last few years it has become clear that polycystic kidney diseases (PKDs) are ciliopathies, due to mutations in cilia related genes. In this proposal we will study two apparently recessively inherited PKDs, autosomal recessive PKD (ARPKD) and Meckel syndrome (MKS). In ARPKD, the disease is largely restricted to the kidney and liver, while MKS is a lethal, syndromic PKD in which central nervous system and digital defects are typically also found. ARPKD is considered a genetically homogenous disorder due to PKHD1 mutation, although studies of large populations with an ARPKD-like phenotype have found PKHD1 mutations in only about one half of individuals. MKS is genetically more complex with 11 genes identified and alleles at other loci also thought to play a role; possible oligogenic inheritance. Although on the surface these diseases appear very different, recent publications and our preliminary data suggest genetic overlap. Hypomorphic mutations in the autosomal dominant PKD (ADPKD) gene, PKD1, have been shown to cause an ARPKD-like disease and exon enrichment and NGS has shown PKHD1, and the related PKHDL1, alleles in MKS patients. The purpose of this study is to employ cutting-edge genetic methods to understand the full genetic load in these two disorders, while animal models will be utilized to determine the significance of detected variants, plus the strength of epistatic effects between ciliopathy genes. In the first aim a total of ~100 MKS and ~200 ARPKD-like families, that are genetically unresolved, will be analyzed by exon enrichment and next-generation sequencing of ciliopathy and other ciliogenes, plus, when appropriate, whole exome analysis. Variants will be assessed bioinformatically and in Aim 2 cellular systems and the model organisms C. elegans and zebrafish employed to assess the significance of the most promising variants. The concept that the phenotype found in MKS and some ARPKD-like families is due to alleles at more than one gene, that epistasis plays a central role, will be tested in Aim 3 employing C. elegans, zebrafish and interbreeding of mouse models of ciliopathies. The final aim will explore the role of the PKHD1 paralog and suspected ciliogene, PKHDL1, by developing knock- in/out mouse models to characterize expression and the phenotype associated with disruption of all transcripts. Together these studies will provide a much clearer view of the etiology of ARPKD-like disease and MKS and clarify the true complexity of these "simple" genetic diseases. PUBLIC HEALTH RELEVANCE: Polycystic kidney diseases (PKD) are a group of inherited disorders that result in cyst development in the kidney resulting in renal failure and a range of morbidities beyond the kidney. In this grant we will study two severe forms of PKD, ARPKD and Meckel syndrome (MKS) that result in utero and neonatal disease, and often result in death. These have been considered recessively inherited disorders but with new genetic tools we are uncovering more genetic complexity. Here we will determine the genetic cause of these disorders, not only in terms of the disease causing mutations but also identifying other genes that may modify the way the disease presents and progresses. Various cellular and model animal systems will be employed to determine the importance of DNA variants identified and genetic interaction between genes. These studies will be of diagnostic and prognostic importance in these disorders and help to understand the true complexity of simple genetic diseases.

描述（由申请人提供）：在过去的几年中，由于纤毛相关基因的突变，多囊性肾脏疾病（PKD）是纤毛病。在此提案中，我们将研究两个显然遗传遗传的PKD，即常染色体隐性PKD（ARPKD）和Meckel综合征（MKS）。在ARPKD中，该疾病在很大程度上仅限于肾脏和肝脏，而MK是一种致命的，综合症PKD，通常还发现中枢神经系统和数字缺陷。尽管PKHD1突变引起的ARPKD被认为是遗传均质疾病，尽管对具有ARPKD样表型的大种群的研究发现了仅一半个个体的PKHD1突变。 MK在遗传上更为复杂，有11个基因，而在其他基因座的等位基因也被认为起作用。可能的寡原性继承。尽管从表面上看，这些疾病的表现非常不同，但最近的出版物和我们的初步数据表明遗传重叠。已证明常染色体显性PKD（ADPKD）基因PKD1中的低形态突变会引起类似ARPKD的疾病和外显子富集，而NGS显示了PKHD1，而相关的PKHDL1则显示了MKS患者的等位基因。这项研究的目的是采用尖端的遗传学方法来了解这两种疾病中的全部遗传负荷，而动物模型将被用来确定检测到的变体的重要性，以及纤毛病基因之间上皮作用的强度。在第一个目的中，总共有算在遗传上没有解决的〜100 mks和〜200个类似ARPKD的家族，将通过外显子富集和下一代纤毛病和其他纤毛基因分析，并在适当的情况下进行整体外质分析。将通过生物信息和AIM 2细胞系统评估变体，以及用于评估最有希望变体的重要性的秀丽隐杆线虫和斑马鱼的模型。 MK和某些类似ARPKD的家族中发现的表型的概念是由于等位基因以上的基因而引起的，Epitisasis扮演着核心角色，将在AIM 3中测试，使用秀丽隐杆线虫，斑马鱼和纤毛病小鼠模型的杂交。最终目的将通过开发敲入/输出小鼠模型来表征表达和与所有转录本的破坏相关的表征表征来探讨PKHD1旁系同源物和可疑的纤毛pKHDL1的作用。这些研究共同将对ARPKD样疾病和MK的病因提供更清晰的看法，并阐明这些“简单”遗传疾病的真正复杂性。 公共卫生相关性：多囊性肾脏疾病（PKD）是一组遗传性疾病，导致肾脏的囊肿发育，导致肾衰竭和肾脏以外的一系列病态。在这笔赠款中，我们将研究两种严重的PKD，ARPKD和Meckel综合征（MKS），它们导致子宫和新生儿疾病，并经常导致死亡。这些被认为是隐性遗传性疾病，但是使用新的遗传工具，我们正在发现更加遗传复杂性。在这里，我们将确定这些疾病的遗传原因，不仅在引起突变的疾病方面，而且还鉴定出可能改变疾病表现方式的其他基因。将采用各种细胞和模型动物系统来确定基因之间鉴定出的DNA变异和遗传相互作用的重要性。这些研究将在这些疾病中具有诊断和预后的重要性，并有助于了解简单遗传疾病的真正复杂性。