Genetic and Functional Dissection of Congenital Anomalies of the Brain

大脑先天性异常的遗传和功能解剖

• 批准号: 9752755
• 负责人: Erica Ellen Davis
• 金额: $ 25.04万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9752755
• 关键词: Ablation; Affect; Anatomy; Anencephaly; Animal Model; Antisense Oligonucleotides; Award; Bioinformatics; Biological Assay; Biological Models; Birth; Brain; CRISPR/Cas technology; Candidate Disease Gene; Cell physiology; Clinical; Clinical Research; Complement; Complex; Congenital Abnormality; Congenital cerebellar hypoplasia; Congenital cerebral hernia; Congenital neurologic anomalies; Counseling; Data; Defect; Development; Diagnosis; Diagnostic; Dideoxy Chain Termination DNA Sequencing; Disease; Disease model; Dissection; Embryo; Enrollment; Etiology; Family; Fetus; Funding; Future; Genes; Genetic; Genetic study; Genome; Genomics; Genotype; Healthcare Systems; Heterogeneity; Holoprosencephaly; Human; Hybrids; Image; Investigation; Karyotype; Knowledge; Larva; Lead; Lesion; Light; Literature; Magnetic Resonance Imaging; Messenger RNA; Microcephaly; Modeling; Molecular; Molecular Diagnosis; Mutate; Nervous system structure; Neuraxis; Neurological observations; Newborn Infant; Nucleotides; Optics; Organ; Orthologous Gene; Parents; Pathogenicity; Phenotype; Physicians; Physiological; Population Genetics; Positioning Attribute; Pregnancy; Prenatal Diagnosis; Preventive; Process; Proteins; Recurrence; Resolution; Risk; Schizencephaly; Scientist; Signal Pathway; Structure; Syndrome; System; Technology; Testing; Therapeutic; Translating; Translational Research; Ultrasonography; United States; Variant; Work; Zebrafish; base; body system; brain abnormalities; brain malformation; causal variant; clinical Diagnosis; clinical care; clinical phenotype; exome; exome sequencing; experience; fetal; gene discovery; genetic analysis; genetic disorder diagnosis; genetic pedigree; genetic variant; genome editing; genome-wide; genomic data; human disease; human model; improved; in vivo; in vivo Model; innovation; insertion/deletion mutation; knock-down; lissencephaly; multidisciplinary; nervous system development; novel; outcome forecast; perinatal period; phenotypic data; prenatal; prospective; rare variant; therapeutic target; tool; zebrafish genome

PROJECT SUMMARY Human brain development remains an incompletely understood process, yet congenital abnormalities of this complex structure affect approximately 3/1,000 pregnancies and more than 2000 newborns annually in the United States, posing a substantial burden on the health care system. Congenital brain abnormalities, hallmarked by vast phenotypic heterogeneity, include but are not limited to holoprosencephaly, schizencephaly, anencephaly, encephalocele, microcephaly, ventriculomegaly, cerebellar hypoplasia, and disorders of cortical development, such as lissencephaly. The paired approach of: (1) prenatal diagnosis using a combination of ultrasound and fetal MRI to characterize aberrant phenotypes; with (2) genetic analysis to determine causal lesions, has greatly improved the ability to accurately counsel families about diagnosis, prognosis, and recurrence risk. More recently, prenatal whole exome sequencing (WES) has been applied in cases of lethal or multiple fetal abnormalities to make a molecular diagnosis that otherwise could not be identified with traditional testing. Pilot data from our group and others using WES show a diagnostic rate of 16- 30% in cases of multiple fetal abnormalities, but only 1-2% in isolated brain abnormalities, indicating a critical need to improve diagnostic capabilities and identify novel genes critical to human brain development. We posit that the overabundance of unresolved fetal cases is in large part due to: (1) a knowledge gap in our understanding of the repertoire of genotypes underlying brain abnormalities with prenatal onset; and (2) limitations of population genetics to establish causality of rare variants in novel candidate genes. Here, two CTSA-funded teams who are at the forefronts of prenatal genetic diagnostics and in vivo zebrafish modeling of human disease, at UNC and Duke, respectively, will team up to overcome the current challenges of diagnosing brain abnormalities with a prenatal onset. We will intersect exome- and genome-wide variation data with experimentally tractable and relevant model systems, zebrafish (Danio rerio). We hypothesize that bioinformatics filters using prenatal WES data will reveal novel candidate genes, which can be applied to a zebrafish model to generate initial discoveries critical to human brain development and translate into improved clinical care. First, we will perform bioinformatic analysis of 10 clinically ascertained fetuses with CNS anomalies and their parents using a tiered filtering strategy; and we will apply this analysis paradigm iteratively to 32 prospectively enrolled fetuses and their families. Second, we will establish relevance of candidate genes to brain development and determine variant pathogenicity using state-of-the-art genome editing and phenotyping tools in zebrafish. Completion of our work will expand our understanding of the molecular processes governing prenatal brain development; establish a clinical-research hybrid platform readily applicable to other anatomical organ defects detectable by fetal imaging; and build a suite of animal models of aberrant CNS development with potential for future use in therapeutic target identification.

项目摘要 人类大脑的发育仍然是一个不完全理解的过程，但这种先天性异常， 复杂的结构每年影响大约3/1 000的怀孕和2 000多名新生儿， 这对美国的医疗保健系统造成了巨大的负担。先天性大脑异常， 以巨大表型异质性为特征，包括但不限于前脑无裂畸形， 无脑畸形、无脑畸形、脑膨出、小头畸形、脑室扩大、小脑发育不全和 皮质发育障碍，如无脑畸形。配对方法：（1）产前诊断， 超声和胎儿MRI的组合，以表征异常表型;（2）遗传分析， 确定因果病变，大大提高了准确咨询家庭诊断的能力， 预后和复发风险。最近，产前全外显子组测序（WES）已经应用于 致命的或多个胎儿异常的情况下，作出分子诊断，否则不能 与传统的测试。我们小组和其他使用WES的试验数据显示，诊断率为16- 30%的情况下，多个胎儿异常，但只有1-2%，在孤立的脑异常，表明一个关键的 需要提高诊断能力，并确定对人类大脑发育至关重要的新基因。我们断定 过多的未解决的胎儿病例在很大程度上是由于：（1）我们的知识差距 了解产前发病的脑异常的基因型库;和（2） 群体遗传学在确定新候选基因中罕见变异的因果关系方面的局限性。这里为两个 CTSA资助的团队在产前遗传诊断和体内斑马鱼建模的前沿， 人类疾病，分别在剑桥和杜克，将联手克服目前的挑战，诊断 产前出现的脑部异常我们将把外显子组和全基因组变异数据与 实验上易处理的和相关的模型系统，斑马鱼（Danio rerio）。我们假设 使用产前WES数据的生物信息学过滤器将揭示新的候选基因，这些基因可以应用于 斑马鱼模型，以产生对人类大脑发育至关重要的初步发现，并转化为改进的 临床护理首先，我们将对10例临床确诊的CNS胎儿进行生物信息学分析， 异常及其父母使用分层过滤策略;我们将反复应用此分析范式 32例前瞻性入组胎儿及其家属。其次，我们将建立候选基因的相关性 大脑发育和使用最先进的基因组编辑确定变异致病性， 斑马鱼的表型分析工具。我们工作的完成将扩大我们对分子的理解 控制产前大脑发育的过程;容易建立临床-研究混合平台 适用于胎儿成像可检测的其他解剖器官缺陷;并建立一套动物模型， 异常的CNS发育，未来有可能用于治疗靶点鉴定。