Genetic and Functional Studies of Human Ciliary Syndromes

人类睫状体综合征的遗传和功能研究

基本信息

批准号：
8117848
负责人：
NICHOLAS KATSANIS
金额：
$ 9.64万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-29 至 2011-09-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8117848
关键词：
Acids Attention Bardet-Biedl Syndrome Biogenesis Biological Biological Assay Biology C-terminal Caenorhabditis elegans Carrier Proteins Cell Line Cell model Cell surface Cells Chlamydomonas Chlamydomonas reinhardtii Chromosome Mapping Cilia Classification Collection Coupled Data Data Set Defect Devices Disease Distant Down-Regulation Embryo Epitopes Esthesia Evolution Exhibits Family Flagella Functional disorder Gene Proteins Genes Genetic Genetic Transcription Genome Genomics Golgi Apparatus Heart Hereditary Disease Homo sapiens Human Human Genetics Human Pathology Hydrocephalus In Vitro Kidney Kidney Diseases Lead Left Life Cycle Stages Liquid substance Liver Location Mammalian Cell Maps Messenger RNA Microtubules Molecular Monitor Morphology Movement Mus Mutation Nature Nephronophthisis Nervous system structure Neurons Nodal Oligonucleotides Organ Organelles Organism Outcome Pancreas Patients Pericentriolar Regions Phenotype Photoreceptors Physiological Physiology Process Proteins Proteome RNA Interference Regulation Relative (related person)Reproduction Resources Retina Retinal Degeneration Reverse Transcriptase Polymerase Chain Reaction Role Sensory Sensory Process Signal Transduction Specialist Structure Swelling Syndrome Testing Time Tissues Trachea Transcriptional Regulation Transport Vesicles Transportation Tubulin Ursidae Family base cell motility cell type cohort comparative comparative genomics computer studies computerized tools density disease phenotype early onset expression vector genetic pedigree human disease insight kinetosome loss of function loss of function mutation mammalian genome migration novel patient population protein function sperm cell tool

项目摘要

DESCRIPTION (provided by applicant): Cilia and flagella are ancient, evolutionarily conserved organelles that project from cell surfaces to perform diverse biological roles, including whole-cell locomotion, movement of fluid, chemo-, mechano- and photosensation, and sexual reproduction. Consistent with their stringent evolutionary conservation defects in cilia or flagella are associated with a wide range of human diseases. Loss or dysfunction of nodal cilia perturbs left-right axis determination, whereas sensory cilia defects lead to polycystic liver and kidney disease. Likewise, dysfunction of ependymal cilia cause hydrocephalus, and defective transportation of proteins along the photoreceptor connecting cilium leads to retinal degeneration. Despite their profound importance, cilia and their roles in human physiology have only recently gained broader attention and the fact that most mammalian cells have the capacity to ciliate has been under-appreciated. Here we propose to perform a comprehensive analysis of the mammalian genome and proteome to identify and validate the fraction of human proteins involved in ciliary structure and function, and to determine their contribution to human genetic disease. First, we will use selective evolutionary conservation coupled with microarray analyses to differentiate between proteins necessary for the function of sensory cilia, motile cilia, or both. Second, focusing on proteins expressed in sensory cilia, we will interrogate their direct involvement in ciliary function by determining their subcellular localization in ciliated cells. Third, we will perform high-density mapping of consanguineous families with established ciliation disorders such as Bardet-Biedl syndrome (BBS) and nephronophthisis (NPH) and determine for each pedigree all regions in the genome that display homozygosity by descent. We will then intersect our computational and experimental ciliary proteome data with our genetic mapping information and identify novel BBS and NPH genes. Finally, to investigate the mechanism of dysfunction, we will suppress the mRNA message of novel disease genes in mammalian ciliated cells and model the consequences of loss of function mutations in ciliary morphology, intraflagellar transport and transcriptional regulation during the ciliary life cycle. These studies will further significantly our understanding of the function of the cilium, one of the least-studied cellular organelles, yet of major physiological importance, and provide novel tools to dissect the molecular basis of human genetic disease.

描述（由申请人提供）：纤毛和鞭毛是古老的、进化上保守的细胞器，它们从细胞表面突出来执行多种生物学作用，包括全细胞运动、液体运动、化学、机械和光感以及有性生殖。与严格的进化保护一致，纤毛或鞭毛的缺陷与多种人类疾病相关。淋巴结纤毛的缺失或功能障碍会扰乱左右轴的确定，而感觉纤毛缺陷会导致多囊肝和肾病。同样，室管膜纤毛功能障碍会导致脑积水，而沿着连接纤毛的光感受器的蛋白质运输缺陷会导致视网膜变性。尽管纤毛极其重要，但它们在人类生理学中的作用直到最近才获得更广泛的关注，而且大多数哺乳动物细胞具有纤毛能力的事实尚未得到充分认识。在这里，我们建议对哺乳动物基因组和蛋白质组进行全面分析，以识别和验证参与纤毛结构和功能的人类蛋白质部分，并确定它们对人类遗传疾病的贡献。首先，我们将使用选择性进化保守与微阵列分析相结合来区分感觉纤毛、运动纤毛或两者功能所需的蛋白质。其次，重点关注感觉纤毛中表达的蛋白质，我们将通过确定它们在纤毛细胞中的亚细胞定位来探究它们对纤毛功能的直接参与。第三，我们将对患有纤毛疾病（例如 Bardet-Biedl 综合征 (BBS) 和肾结核 (NPH)）的近亲家族进行高密度作图，并确定每个谱系的基因组中显示血统纯合性的所有区域。然后，我们将计算和实验的纤毛蛋白质组数据与我们的遗传图谱信息相交叉，并识别新的 BBS 和 NPH 基因。最后，为了研究功能障碍的机制，我们将抑制哺乳动物纤毛细胞中新疾病基因的 mRNA 信息，并模拟纤毛生命周期中纤毛形态、鞭毛内运输和转录调控功能突变丧失的后果。这些研究将进一步加深我们对纤毛功能的理解，纤毛是研究最少的细胞器之一，但具有重要的生理重要性，并提供新的工具来剖析人类遗传疾病的分子基础。