Models to study heritable and de novo DAND mutations

研究遗传性和从头 DAND 突变的模型

• 批准号: 10348921
• 负责人: Philip J. Jensik
• 金额: $ 7.38万
• 依托单位: SOUTHERN ILLINOIS UNIVERSITY CARBONDALE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-17 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10348921
• 关键词: Adult; Affect; Alleles; Angelman Syndrome; Behavior; Biological Models; Brain; CRISPR/Cas technology; Cognitive deficits; Complex; DNA; Data; Development; Disease; Dominant-Negative Mutation; Engineering; Fright; Gene Expression; Generations; Genes; Genetic Transcription; Genomics; Genotype; Goals; Heritability; Hippocampus (Brain); Human; Individual; Intellectual functioning disability; Knock-in; Knock-in Mouse; Knock-out; Knockout Mice; Learning; Mediating; Memory; Memory impairment; Messenger RNA; Modeling; Molecular; Mus; Mutation; Neurodevelopmental Disorder; Pathogenesis; Patients; Phenotype; Point Mutation; Proteins; Research; Rett Syndrome; Synaptic plasticity; Testing; Transcriptional Activation; Variant; autism spectrum disorder; base; clinical phenotype; cognitive function; conditional knockout; de novo mutation; developmental disease; differential expression; experimental study; functional outcomes; gain of function; gene repression; homologous recombination; innovation; insight; loss of function; morris water maze; mouse model; mutant; nestin protein; neurobehavioral; neurobehavioral test; postnatal; severe intellectual disability; therapeutic development; transcription factor; transcriptome sequencing; transcriptomics

Project Summary DEAF1-associated neurodevelopmental disorder (DAND) is a phenotypic spectrum of features that include intellectual disability and autism spectrum disorder (ASD). Individuals with DAND have clinical phenotypes that overlap with other neurodevelopmental disorders including Angelman and Rett syndromes. DAND is caused by single allelic de novo and bi-allelic heritable mutations in the DEAF1 gene. Our functional studies indicate that identified de novo mutations disrupt DEAF1 transcriptional activity and suggest that the mutations are dominant negative. The effects of heritable variants on DEAF1 function remain unclear. This proposal focuses on two identified DEAF1 variants, p.Q264P (de novo) and p.R226W (heritable). The resultant spectrum disorders of individuals with heritable and de novo DEAF1 mutations is remarkably quite similar. We hypothesize there is a common set of altered DEAF1 target genes, caused by the heritable and de novo DEAF1 mutations, that underlie the pathogenesis of DAND. The innovation of this proposal is the development of new model systems that more closely represent DAND. The current best model for assessing the effects of decreased DEAF1 activity on cognitive function is a conditional knockout model. Although the knockout model demonstrates the importance of DEAF1 expression in learning and memory, it does not mirror the allelic profile of DAND or, more importantly, account for dominant negative or possible gain-of-function actions of the mutant DEAF1 proteins. We will use CRISPR/Cas9-mediated genomic engineering to develop p.R227W and p.Q265P (analogous to p.R226W and p.Q264P in humans) point mutation knock-in mouse models of DAND. Neurobehavioral testing will assess the effects of mutant DEAF1 proteins on learning and memory. We will also identify transcriptomic changes in postnatal developing and adult hippocampus. A common set of differentially expressed genes, likely causal to the cognitive deficits, will be determined by comparing these two models. These models will provide important insights into DAND, as well as other developmental disorders with overlapping clinical phenotypes.

项目概要 DEAF1 相关神经发育障碍 (DAND) 是一系列表型特征，包括 智力障碍和自闭症谱系障碍（ASD）。 DAND 患者的临床表型 与其他神经发育障碍重叠，包括天使综合征和雷特综合征。 DAND 是由 DEAF1 基因中的单等位基因从头突变和双等位基因可遗传突变。我们的功能研究表明 鉴定出的从头突变会破坏 DEAF1 转录活性，表明这些突变是显性突变 消极的。可遗传变异对 DEAF1 功能的影响仍不清楚。该提案主要关注两点 鉴定出 DEAF1 变体 p.Q264P（从头）和 p.R226W（可遗传）。由此产生的谱系障碍 具有遗传性 DEAF1 突变和新发 DEAF1 突变的个体非常相似。我们假设有一个 由可遗传和 de novo DEAF1 突变引起的常见一组改变的 DEAF1 靶基因 DAND 的发病机制。该提案的创新之处在于开发了更多新模型系统 密切代表DAND。目前评估 DEAF1 活性降低对健康的影响的最佳模型 认知功能是一个条件敲除模型。尽管淘汰赛模型显示了重要性 DEAF1 在学习和记忆中的表达，它并不反映 DAND 的等位基因特征，或更重要的是， 解释突变 DEAF1 蛋白的显性失活或可能的功能获得作用。我们将使用 CRISPR/Cas9介导的基因组工程开发p.R227W和p.Q265P（类似于p.R226W和 p.Q264P（人类）DAND 点突变敲入小鼠模型。神经行为测试将评估 突变 DEAF1 蛋白对学习和记忆的影响。我们还将鉴定转录组变化 产后发育和成年海马体。一组常见的差异表达基因，可能导致 认知缺陷将通过比较这两个模型来确定。这些模型将提供重要的 对 DAND 以及具有重叠临床表型的其他发育障碍的见解。