Molecular Analysis of primary cilia proteins in human development

人类发育中初级纤毛蛋白的分子分析

• 批准号: 10594444
• 负责人: Rolf W Stottmann
• 金额: $ 38.5万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10594444
• 关键词: Address; Award; Biochemistry; Cell physiology; Cellular biology; Cilia; Congenital Abnormality; Data; Development; Disease; Dysmorphology; Embryology; Ethylnitrosourea; Genes; Genetic; Genetic study; Goals; Health; Human; Human Development; Human Genetics; Intellectual functioning disability; Intervention; Knowledge; Laboratories; Literature; Microcephaly; Mission; Molecular; Molecular Analysis; Mus; Mutagenesis; Mutant Strains Mice; Neurons; Obesity; Organ; Pathology; Phenotype; Physiology; Proteins; Public Health; Quantitative Trait Loci; Research; Role; Severities; Signal Transduction; Tissues; United States National Institutes of Health; Work; burden of illness; ciliopathy; craniofacial; experience; gene interaction; genetic analysis; human disease; human genomics; insight; malformation; mouse genetics; novel; patient population; trafficking; Address; Award; Biochemistry; Cell physiology; Cellular biology; Cilia; Congenital Abnormality; Data; Development; Disease; Dysmorphology; Embryology; Ethylnitrosourea; Genes; Genetic; Genetic study; Goals; Health; Human; Human Development; Human Genetics; Intellectual functioning disability; Intervention; Knowledge; Laboratories; Literature; Microcephaly; Mission; Molecular; Molecular Analysis; Mus; Mutagenesis; Mutant Strains Mice; Neurons; Obesity; Organ; Pathology; Phenotype; Physiology; Proteins; Public Health; Quantitative Trait Loci; Research; Role; Severities; Signal Transduction; Tissues; United States National Institutes of Health; Work; burden of illness; ciliopathy; craniofacial; experience; gene interaction; genetic analysis; human disease; human genomics; insight; malformation; mouse genetics; novel; patient population; trafficking

The goal of the research in my laboratory is to study the genetic basis of human craniofacial and CNS malformations. Our efforts in both human and mouse genetics over the past several years have continually directed us towards the primary cilium as a critical hub in signaling for human health and disease. Ciliopathies are diseases associated with both severe congenital malformations as well as nonlethal craniofacial dysmorphology, intellectual disability and obesity (among other conditions). It is clear from the literature that modifying loci are crucial components in understanding much of human disease, but is especially true of the ciliopathies. The focus of this proposal is largely on the primary cilia gene tetratricopeptide repeat domain 21B (Ttc21b). Ttc21b homozygous mouse mutants have several striking features on their own but our preliminary data and the work of others clearly show that TTC21B is a hub in a human ciliopathy network. We have taken four of these candidate interactions from human genetics and recreated them in mouse. All four genes interact with Ttc21b but the cellular and mechanisms of the resulting phenotypes are not yet elucidated. We have also identified multiple novel interacting loci with a combination of ENU mutagenesis and a QTL analysis of the genetic background effects on the severity of the microcephaly phenotype. Thus, we have significant experience in the field and have identified four crucial gaps in knowledge we will address with the support of this MIRA award: 1) How does Ttc21b have such tissue specific effects on organ physiology and developmental signaling, 2) What are the genetic interactors and modifiers of Ttc21b which alter these ciliopathy phenotypes, 3) what is the cellular function of Ttc21b inside the primary cilium, 4) what is the role of Ttc21b outside the cilium. We will use a combination of genetics, molecular embryology, cell biology and biochemistry to address these topics. Many of the ciliary genes are identified to have roles within the primary cilium, but any function outside the cilium has not been elucidated. Identification of such roles would have very a significant effect on the field. Our favorite hypothesis based on preliminary data is that Ttc21b has significant roles in neuronal trafficking. It is clear that a better understanding of human disease will require knowledge of modifying loci and the underlying mechanism(s). Ttc21b is ripe for exploration as a crucial component in a ciliopathy genetic network. A combination of mouse embryology and cell biology inspired by human genomics is an ideal entry point. Our work is likely to not only contribute to knowledge about ciliopathies but point the way forward as a general experimental paradigm for a number of different pathophysiological contexts.

我的实验室研究的目的是研究人类颅面和 中枢神经系统畸形。在过去的几年中，我们在人类和老鼠遗传学方面的努力 一直将我们引向主要纤毛，作为人类信号的关键枢纽 健康与疾病。纤毛病是与严重先天性相关的疾病 畸形以及非致死性颅面畸形，智力残疾和肥胖症 （除其他条件外）。从文献来看，修改基因座至关重要 了解大部分人类疾病的组成部分，但尤其是纤毛病。 该提案的重点主要放在主要的纤毛基因四肽重复域 21B（TTC21B）。 TTC21B纯合小鼠突变体独自具有几个引人注目的功能 我们的初步数据和其他人的工作清楚地表明，TTC21B是人类的枢纽 纤毛病网络。我们从人类遗传学和 用鼠标重新创建它们。所有四个基因都与TTC21B相互作用，但细胞和机制 所得的表型尚未阐明。我们还确定了多种小说 与ENU诱变和遗传分析的QTL分析相互作用的基因座 背景对小头畸形表型严重程度的影响。因此，我们有很大的 在该领域的经验，并确定了我们将解决的四个关键差距 该MIRA奖的支持：1）TTC21B如何对器官产生这种特异性影响 生理和发育信号，2） TTC21b改变了这些纤毛病表型，3）TTC21B的细胞功能是什么 原发性纤毛，4）纤毛以外的TTC21B的作用是什么？我们将结合 遗传学，分子胚胎学，细胞生物学和生物化学来解决这些主题。许多 睫状基因被确定为在主要纤毛内具有作用，但是外部的任何功能 纤毛尚未阐明。识别此类角色将具有非常重要的 对场的影响。我们最喜欢的基于初步数据的假设是TTC21B具有 在神经元贩运中的重要作用。显然，对人类疾病有更好的了解 将需要了解修改基因座和基本机制的知识。 TTC21B已成熟 在纤毛病遗传网络中探索是至关重要的成分。小鼠的组合 受人基因组学启发的胚胎学和细胞生物学是理想的入口点。我们的工作可能是 不仅有助于有关纤毛病的知识，而且还要指出前进的道路 实验范式，用于许多不同的病理生理环境。