A genetic approach to defining the Ttc21b interactome in mammalian ciliopathies

定义哺乳动物纤毛病中 Ttc21b 相互作用组的遗传学方法

基本信息

批准号：
9415039
负责人：
Rolf W Stottmann
金额：
$ 30.03万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-02-01 至 2020-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9415039
关键词：
Address Adult Affect Alleles Area Bioinformatics Biology Candidate Disease Gene Cells Cellular Membrane Cilia Cilium Microtubule Congenic Strain Congenital Abnormality Craniofacial Abnormalities Data Defect Developmental Biology Disease Embryo Ethylnitrosourea Face Felis catus Functional disorder Gene Proteins Gene-Modified Genes Genetic Genetic Techniques Genetic study Goals Human Human Genetics In Vitro Inbred Strains Mice Incidence Interphase Cell Intervention Kidney Knockout Mice Knowledge Lead Lung Meiotic Recombination Microcephaly Microtubules Mission Modeling Molecular Mouse Strains Mus Mutagenesis Mutate Mutation Nervous system structure Organ Organelles Organogenesis Outcome Pathology Pathway interactions Patients Perinatal mortality demographics Phenotype Population Prosencephalon Protein Binding Domain Proteins Public Health Publishing Quantitative Trait Loci Research Scaffolding Protein Severities Signal Transduction Skeleton Solid Testing Therapeutic Intervention Tissues United States National Institutes of Health Work base burden of illness ciliopathy cohort craniofacial craniofacial tissue gene product genetic approach in vivo innovation insight interest mouse model mutant next generation sequencing novel patient population positional cloning public health relevance scaffold trafficking

项目摘要

DESCRIPTION (provided by applicant): Ciliopathies are a spectrum of diseases resulting from defects in primary cilia function affecting 1:800 people. Primary cilia are microtubule based organelles found on almost all cells and crucial for proper signal transduction of a number of molecular pathways. Ciliopathies affect a wide range of tissues including the nervous system, craniofacial tissues, skeleton, kidneys, lungs and digestive organs. These manifest as both congenital and adult-onset defects. Genetic studies of ciliopathy patients show TTC21B (tetratricopeptide repeat domain-containing protein1B) is the most commonly mutated cilia gene identified to date. In addition to defects associated with loss of just TTC21B, mutations in trans with a number of other ciliary genes lead to ciliopathies. We have recently shown loss of Ttc21b in the mouse leads to perinatal lethality and organogenesis defects. We also note some of these phenotypes are dependent on the specific inbred mouse strain background. The TTC21B protein is large with many protein-protein interaction domains and important for intraflagellar transport and regulating signal transduction in the cilium. All of these data together lead us to the central hypothesis that TTC21B serves as a network hub for scaffolding and trafficking activities essential for proper cilia form and function. The goal of this application is identify genes and proteins interacting with Ttc21b: the Ttc21b "interactome," and begin to understand how these interact in the cell. The rationale for the project is that a more complete understanding of how TTC21B acts is likely to give insight to a range of ciliopathies. We will address this hypothesis and achieve these goals with the following three specific aims: 1) identify chromosomal regions containing genes modifying the Ttc21bnull/null phenotype in the B6 and FVB mouse strains, 2) identify novel genetic interactions with Ttc21b using a forward genetic approach, and 3) study functional mechanisms of genes interacting with Ttc21b. The first aim will utilize a QTL analysis to identify loci regulating the strain specific phenotypes we see in Ttc21bnull/null embryos. The second aim will take a forward genetic, ENU mutagenesis approach to identify novel interactions with TTC21B in an unbiased manner. The third aim will recapitulate interactions identified in humans or previously identified in mouse. After verifying these interactions yield ciliopathy phenotype(s), we will perform further analyses in vitro and in vivo to study the molecular mechanisms of ciliary dysfunction. These studies will focus on ciliary trafficking and Shh signal transduction. The significance of this project is that these studies wil together dramatically increase our understanding of how TTC21B acts within the primary cilium and why perturbation of function leads to ciliopathic disease. These studies will fill an important gap in our knowledge and identify possible areas for therapeutic intervention. These advances are not specific to TTC21B but are likely going to be largely applicable to multiple areas of primary cilia biology inter- est. The innovation of this project lies in the application of unbiase genetic techniques to identify the Ttc21b interactome in close concert with solid molecular studies to determine the underlying mechanism(s).

描述（由申请人提供）：纤毛病是由初级纤毛功能缺陷引起的一系列疾病，影响 1:800 人。初级纤毛是几乎所有细胞中都存在的基于微管的细胞器，对于许多分子途径的正确信号转导至关重要。纤毛病影响广泛的组织，包括神经系统、颅面组织、骨骼、肾脏、肺和消化器官。这些表现为先天性和成人发病的缺陷。对纤毛病患者的遗传学研究表明，TTC21B（含四肽重复结构域的蛋白 1B）是迄今为止发现的最常见的纤毛突变基因。除了与 TTC21B 缺失相关的缺陷之外，许多其他纤毛基因的反式突变也会导致纤毛病。我们最近发现小鼠中 Ttc21b 的缺失会导致围产期死亡和器官发生缺陷。我们还注意到其中一些表型取决于特定的近交小鼠品系背景。 TTC21B 蛋白很大，具有许多蛋白质-蛋白质相互作用结构域，对于鞭毛内运输和调节纤毛中的信号转导很重要。所有这些数据共同引导我们得出一个中心假设：TTC21B 作为支架和贩运活动的网络枢纽，对于正确的纤毛形式和功能至关重要。此应用的目标是识别与 Ttc21b 相互作用的基因和蛋白质：Ttc21b“相互作用组”，并开始了解它们如何在细胞中相互作用。该项目的基本原理是，更全面地了解 TTC21B 的作用可能有助于深入了解一系列纤毛病。我们将通过以下三个具体目标来解决这一假设并实现这些目标：1）在 B6 和 FVB 小鼠品系中鉴定含有修饰 Ttc21bnull/null 表型的基因的染色体区域，2）使用正向遗传方法鉴定与 Ttc21b 的新遗传相互作用，3) 研究与 Ttc21b 相互作用的基因的功能机制。第一个目标是利用 QTL 分析来识别调节菌株特定表型的基因座。参见 Ttc21bnull/null 胚胎。第二个目标将采用正向遗传、ENU 诱变方法，以公正的方式识别与 TTC21B 的新相互作用。第三个目标将概括在人类中发现的或先前在小鼠中发现的相互作用。在验证这些相互作用产生纤毛病表型后，我们将在体外和体内进行进一步的分析，以研究纤毛功能障碍的分子机制。这些研究将重点关注纤毛运输和嘘信号转导。该项目的意义在于，这些研究将共同极大地增进我们对 TTC21B 如何在初级纤毛中发挥作用以及功能扰动为何会导致纤毛病的理解。这些研究将填补重要的空白我们的知识差距并确定治疗干预的可能领域。这些进展并不特定于TTC21B，但很可能在很大程度上适用于初级纤毛生物学兴趣的多个领域。该项目的创新在于应用无偏差遗传技术来识别与固体分子密切相关的Ttc21b相互作用组。研究以确定潜在机制。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Gpr63 is a modifier of microcephaly in Ttc21b mouse mutants.

Gpr63 是 Ttc21b 小鼠突变体小头畸形的修饰因子。

DOI：
发表时间：
2019
期刊：
影响因子：
4.5
作者：
Snedeker, John;Gibbons Jr, William J;Paulding, David F;Abdelhamed, Zakia;Prows, Daniel R;Stottmann, Rolf W
通讯作者：
Stottmann, Rolf W