Disease Gene Discovery in Structural Brain Disorders

脑结构疾病中的疾病基因发现

基本信息

批准号：
8674761
负责人：
Kaya Bilguvar
金额：
$ 80.41万
依托单位：
YALE UNIVERSITY
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：
2014-04-05 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8674761
关键词：
Affect Biocompatible Materials Bioinformatics Biological Biology Biopsy Brain Brain Diseases Candidate Disease Gene Cell Culture Techniques Cell Line Cerebrum Code Collaborations Complex Consanguinity Cost Savings DNA Sequence Data Development Disease Drosophila genus Drosophila melanogaster Electroporation Embryo Event Family Fibroblasts Funding Future Genes Genetic Genetic Heterogeneity Genetic screening method Genomics Grant Health Hereditary Disease Heterogeneity Human In Situ Hybridization In Vitro Individual Lead Meleagris gallopavo Modeling Molecular Molecular Profiling Mus Mutation Nature Neurodevelopmental Disorder Pathway interactions Patients Pattern Phenotype Principal Investigator Proteins Public Health Publishing RNA Interference Research Sampling Savings Skin Small Interfering RNA Staging Stimulus System Technology Time Transfection United States National Institutes of Health Variant base brain malformation cohort cost effective exome sequencing experience fetal gene discovery genetic pedigree genetic technology in utero in vivo indexing induced pluripotent stem cell insight kindred member mouse model mutant neurodevelopment next generation next generation sequencing novel pleiotropism programs protein function research study sample collection screening segregation success

Affect Biocompatible Materials Bioinformatics Biological Biology Biopsy Brain Brain Diseases Candidate Disease Gene Cell Culture Techniques Cell Line Cerebrum Code Collaborations Complex Consanguinity Cost Savings DNA Sequence Data Development Disease Drosophila genus Drosophila melanogaster Electroporation Embryo Event Family Fibroblasts Funding Future Genes Genetic Genetic Heterogeneity Genetic screening method Genomics Grant Health Hereditary Disease Heterogeneity Human In Situ Hybridization In Vitro Individual Lead Meleagris gallopavo Modeling Molecular Molecular Profiling Mus Mutation Nature Neurodevelopmental Disorder Pathway interactions Patients Pattern Phenotype Principal Investigator Proteins Public Health Publishing RNA Interference Research Sampling Savings Skin Small Interfering RNA Staging Stimulus System Technology Time Transfection United States National Institutes of Health Variant base brain malformation cohort cost effective exome sequencing experience fetal gene discovery genetic pedigree genetic technology in utero in vivo indexing induced pluripotent stem cell insight kindred member mouse model mutant neurodevelopment next generation next generation sequencing novel pleiotropism programs protein function research study sample collection screening segregation success

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项目摘要

DESCRIPTION (provided by applicant): The introduction of next-generation DNA sequencing platforms led to unprecedented opportunities for gene discovery in structural brain disorders (SBDs), resulting in the identification of several novel genes fundamental in human cerebral and cerebellar development. Despite their success, these studies also revealed significant challenges associated with disease gene discovery in these disorders confirming, once again, their locus and phenotypic heterogeneity. Based on our experience that has been successful in identifying several SBD genes over the past several years, we now propose to ascertain additional cases and biological samples, followed by mutational screening for genes previously associated with SBDs. We will perform exome sequencing of the SBD 'mutation negative' cohort to discover novel genes followed by comprehensive studies to investigate these newly identified genes, using bioinformatics analyses to determine their temporal and spatial expression patterns during embryonic brain development, experimentally validating with in situ hybridization and immunostaining and studying the effects of the variants at the protein level, both by using available patient derived material, such as skin fibroblast cultures, as well as over-expression and inhibition experiments. For a limited number of genes, we aim to establish induced pluripotent stem cell lines and use Drosophila melanogaster RNAi lines and in utero electroporation in mouse to further examine the biological consequences of the mutations during neural development. These studies will continue to unravel the molecular pathways that underlie the complex events that result in formation of the human brain, setting the stage for more detailed future biological studies.

描述（由申请人提供）：下一代DNA测序平台的引入导致结构性脑疾病（SBD）的基因发现的前所未有的机会，从而鉴定了人类大脑和小脑发育中的几种新基因。尽管它们取得了成功，但这些研究还揭示了与这些疾病中的疾病基因发现有关的重大挑战，再次证实了它们的基因座和表型异质性。基于我们在过去几年中成功识别几个SBD基因的经验，我们现在建议确定其他病例和生物样品，然后对先前与SBD相关的基因进行突变筛选。 We will perform exome sequencing of the SBD 'mutation negative' cohort to discover novel genes followed by comprehensive studies to investigate these newly identified genes, using bioinformatics analyses to determine their temporal and spatial expression patterns during embryonic brain development, experimentally validating with in situ hybridization and immunostaining and studying the effects of the variants at the protein level, both by using available patient derived material, such as skin成纤维细胞培养物以及过表达和抑制实验。对于有限数量的基因，我们旨在建立诱导的多能干细胞系，并使用果蝇Melanogaster RNAi系和小鼠的子宫内穿孔，以进一步检查神经发育过程中突变的生物学后果。这些研究将继续揭开导致人脑形成的复杂事件的基础的分子途径，为更详细的未来生物学研究奠定了基础。