Human genetics and molecular mechanisms of congenital hydrocephalus

先天性脑积水的人类遗传学和分子机制

• 批准号: 10533644
• 负责人: ENGIN DENIZ
• 金额: $ 50.46万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10533644
• 关键词: Accounting; Affect; Biological; Biological Models; Biology; Birth; Brain; Brain Diseases; CRISPR/Cas technology; Candidate Disease Gene; Case Study; Cerebral Ventricles; Cerebrospinal Fluid; Cerebrospinal fluid shunts procedure; Cilia; Clinical; Clinical Management; Congenital Hydrocephalus; Copy Number Polymorphism; Coupling; DNA sequencing; Data; Development; Diagnostic; Differentiation and Growth; Disease; Embryo; Etiology; Evaluation; Family; Fluid Balance; Frequencies; Future; Gene Expression; Gene Mutation; Genes; Genetic; Genetic Counseling; Genetic Predisposition to Disease; Genomic approach; Heterogeneity; Homologous Gene; Human; Human Genetics; Hydrocephalus; Immunohistochemistry; Impairment; In Situ Hybridization; International; Knowledge; Lead; Live Birth; Measures; Mediating; Methods; Modeling; Molecular; Morbidity - disease rate; Mus; Mutant Strains Mice; Mutation; Nature; Neural Cell Adhesion Molecule L1; Neural Tube Development; Neurodevelopmental Disorder; Neurons; Nucleotides; Ontology; Operative Surgical Procedures; Optical Coherence Tomography; Parents; Pathogenesis; Pathway interactions; Patients; Pattern; Phenotype; Preventive; Rana; Research; SMARCC1 gene; Social support; Stem Cell Development; Testing; Therapeutic; Time; Transgenic Organisms; Validation; Variant; Work; Xenopus; autism spectrum disorder; base; bioinformatics pipeline; case control; cerebrospinal fluid flow; clinical decision-making; cohort; cost effective; de novo mutation; exome sequencing; experience; functional genomics; gene discovery; genetic approach; genetic architecture; genetic pedigree; human fetal brain; improved; in silico; in vivo; insight; knock-down; mortality; mouse model; mutant; nerve stem cell; neurogenesis; next generation; novel; novel diagnostics; novel therapeutics; overexpression; particle; proband; prognostication; recruit; screening; segregation; stem cell fate; stem cell growth; targeted sequencing; tool; ventricular system

PROJECT SUMMARY Congenital hydrocephalus (CH), the primary enlargement of the cerebrospinal (CSF)-filled brain ventricles, affects 1/1000 births and is a major cause of morbidity and mortality. Although ~60% of all CH cases are predicted to have a genetic etiology, known genes account for <5% of CH cases. Significant gaps in our understanding of the molecular pathogenesis of CH impede the development of preventive, diagnostic, and therapeutic measures. Fundamental obstacles to CH gene discovery using traditional genetic approaches include locus heterogeneity, phenotypic complexity, and the sporadic nature of the majority of CH cases. Whole exome sequencing (WES) has the potential to overcome these obstacles and has led to unprecedented opportunities for gene discovery in autism and structural brain disorders. We recently used WES to identify four novel CH genes, accounting for ~10% of studied CH cases (Furey et al., Neuron, 2018). All four genes are required for neural tube development and regulate neural stem cell (NSC) fate. These results implicate impaired neurogenesis, rather than CSF over-accumulation, in the pathogenesis of a significant subset of CH patients, with potentially paradigm-changing diagnostic and therapeutic implications. As many causal CH genes remain undiscovered, our objective here is to utilize a functional genomics approach to discover, validate, and gain mechanistic insight into newly identified CH-causing mutations. Our hypothesis is that WES will identify multiple novel CH genes, many of which will converge on pathways that regulate the NSC development. Based on our experience that has been successful in identifying several CH and structural brain disorder genes over the past several years, we now propose to ascertain additional sporadic CH case-parent trios and Turkish consanguineous familial CH forms and perform WES on our large, well-phenotyped CH cohort to discover novel de novo and transmitted CH gene mutations. This will be followed by analyses to determine the expression patterns of newly identified prioritized genes during mammalian brain development. We will then rapidly and inexpensively functionally screen prioritized CH candidate gene mutations for their ability to recapitulate hydrocephalus using our novel, validated platform that utilizes live Xenopus embryos, CRISPR/Cas9 gene editing, quantitative optical coherence tomography, and real-time CSF particle tracking (Date et al., Sci Rep, 2019, Accepted). For select validated genes, we will establish Xenopus lines to elucidate the biological consequences of human CH mutations on cilia-regulated CSF dynamics and NSC growth/differentiation and patterning. This functional genomics approach will elucidate the genetic architecture of CH, and set the stage for more detailed future biological studies in mouse models beyond the scope of this proposal. Ultimately, such knowledge has the potential to improve clinical management, prognostication, surveillance, and genetic advice; stimulate research into new non-surgical therapies; and improve the quality of our support for CH patients and their families.

项目概要 先天性脑积水 (CH)，即充满脑脊液 (CSF) 的脑室的原发性增大， 影响 1/1000 的新生儿，是发病和死亡的主要原因。尽管约 60% 的 CH 病例是 预计有遗传病因，已知基因占慢性肝炎病例的比例<5%。我们的重大差距 对 CH 分子发病机制的了解阻碍了预防、诊断和治疗的发展 治疗措施。使用传统遗传学方法发现 CH 基因的根本障碍 包括位点异质性、表型复杂性以及大多数 CH 病例的散发性。 全外显子组测序 (WES) 有潜力克服这些障碍，并带来前所未有的成果 自闭症和结构性脑疾病基因发现的机会。我们最近使用 WES 识别了四个 新的 CH 基因，约占研究 CH 病例的 10%（Furey 等人，Neuron，2018）。所有四个基因都是 神经管发育和调节神经干细胞（NSC）命运所需的。这些结果暗示 在 CH 的一个重要亚型的发病机制中，神经发生受损，而不是脑脊液过度积累 患者，具有潜在的改变范式的诊断和治疗意义。与许多因果 CH 基因仍未被发现，我们的目标是利用功能基因组学方法来发现， 验证并获得对新发现的导致 CH 的突变的机制见解。我们的假设是 WES 将识别多个新的 CH 基因，其中许多将集中在调节 NSC 的途径上 发展。根据我们已成功识别出几种CH和结构脑的经验 过去几年的疾病基因，我们现在建议确定其他散发性 CH 病例父母 三人组和土耳其近亲家族 CH 形成并在我们大型、表型良好的 CH 上进行 WES 队列发现新的从头和传播的 CH 基因突变。接下来将进行分析 确定哺乳动物大脑发育过程中新发现的优先基因的表达模式。 然后，我们将快速且廉价地对优先 CH 候选基因突变进行功能筛选 能够使用我们利用活爪蟾胚胎的新型、经过验证的平台来重现脑积水， CRISPR/Cas9 基因编辑、定量光学相干断层扫描和实时脑脊液粒子跟踪 （Date 等人，Sci Rep，2019 年，已接受）。对于选定的经过验证的基因，我们将建立非洲爪蟾品系来阐明 人类 CH 突变对纤毛调节的脑脊液动力学和 NSC 的生物学影响 生长/分化和模式。这种功能基因组学方法将阐明遗传结构 CH，并为未来在小鼠模型中进行更详细的生物学研究奠定了基础，超出了本研究的范围 提议。最终，这些知识有可能改善临床管理、预测、 监测和遗传建议；刺激新的非手术疗法的研究；并提高质量 我们对慢性肝炎患者及其家人的支持。