Forward Genetic Analysis of Congenital Defects in Cortical Circuits and Structure

皮质回路和结构先天性缺陷的正向遗传学分析

基本信息

批准号：
8760797
负责人：
Rolf W Stottmann
金额：
$ 40.3万
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：
2014
资助国家：
美国
起止时间：
2014-05-15 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8760797
关键词：
Adult Affect Alleles Animal Model Behavior Disorders Biological Assay Birth Brain Brain Diseases Cells Childhood Clinical Cloning Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Congenital Abnormality Cortical Malformation Defect Developed Countries Development Diagnostic Disease Ethylnitrosourea Etiology Future Gene Mutation Genes Genetic Genetic Counseling Genome Goals Hand Human Human Genetics Knowledge Lead Mental disorders Molecular Morbidity - disease rate Movement Disorders Mus Mutagenesis Mutate Mutation Neocortex Nervous system structure Neurologic Neurology Neurons Pathology Pathway interactions Patients Pediatric Hospitals Phenotype Population Positioning Attribute Prosencephalon Recurrence Reporter Research Resources Role Structural Congenital Anomalies Structure System Technology Testing Therapeutic Intervention Tissues Transgenic Animals Transgenic Organisms Work World Health Organization base brain malformation clinically relevant developmental neurobiology exome sequencing forward genetics gene correction genetic analysis genetic pedigree hippocampal pyramidal neuron insight interest mortality mouse model nervous system disorder neurodevelopment neuron development next generation sequencing novel patient population positional cloning public health relevance relating to nervous system research study success therapeutic target tool

项目摘要

DESCRIPTION (provided by applicant): The mammalian neocortex is an enormous network of cells, each making thousands of connections and an array of neurological conditions can result from inappropriate cortical structure or connectivity. Many of these congenital brain defects have a genetic origin but we still lack a full understanding of the genes and mechanisms involved. The overall objective of this application is to use forward genetic approaches in mouse and human to identify and validate novel alleles important for development of cortical circuitry and overall structure. Our central hypothesis is that a synergistic and unbiased forward genetic approach in mouse and human will lead to fundamental discoveries in the genetics of cortical circuit formation and structural development. The rationale of this proposed research is that by identifying novel genes through forward genetic approaches which are required for normal cortical development using both human and mouse genetics, we are then positioned to use this information and tools to study the etiological mechanisms of human cortical malformations in subsequent studies. We will test this central hypothesis and accomplish the goals of this application by pursuing the following three specific aims: 1) use forward genetics in the mouse to efficiently generate and capture genetic mutations in loci important for cortical circuit formation and structural development, 2) identify and validate causal mutations in novel mouse models of cortical circuit formation and structural brain defects, and 3) apply next-generation sequencing approaches to identify mutations leading to human movement disorders and structural brain defects. The aims are accomplished by an ENU mutagenesis approach in the mouse with the addition of a novel transgenic reporter which is expressed specifically in cortical layer V pyramidal neurons. The mutations are then cloned and validated through a number of functional studies. The human genetics studies are performed with the application of exome sequencing to carefully selected familial cases of movement disorders and structural brain malformations. These studies will identify several genes essential for mammalian forebrain structure and function. The significance of this work is found in the specific application to cortial circuitry and structure, and that an unbiased approach such as this has the capability to implicate entirely new pathways in neurological disease. A synergistic approach using both mouse and human genetics to specifically query these aspects of neural development allows fundamental insights into the genetics of development and disease. Such knowledge is not only critical to further understand the basic mechanisms of neurodevelopment, but also has immediate clinical relevance through identification of a number of potential therapeutic targets. Furthermore, these mouse models provide a reusable resource to directly characterize the role of the mutated gene in neurodevelopment, and potentially serve as a tool to test future therapeutic interventions. Taken together, these findings are therefore applicable to basic developmental neurobiology, pediatric and adult neurology, human genetics and genetic counseling.

描述（由申请人提供）：哺乳动物新皮层是一个巨大的细胞网络，每种都建立了数千个连接和一系列神经系统条件，可能是由不适当的皮质结构或连通性造成的。这些先天性大脑缺陷中有许多具有遗传来源，但我们仍然对所涉及的基因和机制有充分的了解。该应用的总体目的是使用小鼠和人类中的前瞻性遗传方法来识别和验证对皮质回路和整体结构发展重要的新颖等位基因。我们的中心假设是，小鼠和人类中的一种协同和公正的前向遗传方法将导致皮质回路形成和结构发育的遗传学中的基本发现。这项拟议的研究的基本原理是，通过使用人和小鼠遗传学的正常皮质发育所必需的远期遗传学方法来识别新的基因，然后我们可以使用这些信息和工具来研究人类皮质畸形的病因机制，以期在后续研究中。我们将通过追求以下三个特定目的来检验这一中心假设，并实现此应用的目标：1）使用鼠标中的前遗传学来有效地产生和捕获基因座中对皮质电路形成和结构发展重要的基因遗传突变，2）确定和验证在识别的小鼠脑模型中，并识别和验证人类介绍的新型脑部模型，并识别结构性的脑部模型，并识别结构性的脑部模型，并识别结构性的脑部模型，3）和结构性大脑缺陷。目的是通过在小鼠中的ENU诱变方法来实现的，并添加了新型的转基因报告基因，该报告特异性在皮质层V锥体神经元中表达。然后，通过许多功能研究对突变进行克隆和验证。人类遗传学研究是通过在精心选择的运动障碍和结构性脑畸形的家族性病例中应用的。这些研究将确定几种对哺乳动物前脑结构和功能必不可少的基因。这项工作的意义在皮层回路和结构的特定应用中发现，这种无偏见的方法具有在神经系统疾病中牵涉到全新途径的能力。一种使用小鼠和人类遗传学的协同方法专门查询神经发育的这些方面，可以对发展和疾病的遗传学进行基本见解。这种知识不仅对于进一步了解神经发育的基本机制至关重要，而且还通过鉴定许多潜在的治疗靶标具有即时临床相关性。此外，这些小鼠模型提供了可重复使用的资源，可以直接表征突变基因在神经发育中的作用，并有可能作为测试未来治疗干预措施的工具。综上所述，这些发现适用于基本的发育神经生物学，小儿和成人神经病学，人类遗传学和遗传咨询。