Effects of DEAF1 on Neuronal Activity and Target Gene Expression.

DEAF1 对神经元活动和靶基因表达的影响。

• 批准号: 8871064
• 负责人: Philip J. Jensik
• 金额: $ 18.44万
• 依托单位: SOUTHERN ILLINOIS UNIVERSITY CARBONDALE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8871064
• 关键词: Adaptive Behaviors; Affect; Amino Acid Substitution; Area; Biological Assay; Brain; C57BL/6 Mouse; DNA; DNA Binding; DNA Binding Domain; Defect; Dominant-Negative Mutation; EMSA; Eukaryotic Initiation Factor-4G; Gene Expression; Gene Mutation; Gene Targeting; Genes; Genetic Predisposition to Disease; Genetic Transcription; Hippocampus (Brain); Human; Immunofluorescence Immunologic; In Vitro; Individual; Intellectual functioning disability; Knock-out; Learning; Link; Long-Term Depression; Long-Term Potentiation; Measurable; Memory; Missense Mutation; Molecular; Mus; Mutate; Mutation; Neuraxis; Neurons; Outcome; Pathogenesis; Point Mutation; Population; Proteins; RNA; Reporting; Research; Role; Sampling; Serotonin Receptor 5-HT1A; Slice; Testing; Variant; anxiety-like behavior; autism spectrum disorder; base; chromatin immunoprecipitation; cognitive ability; cognitive function; conditioned fear; differential expression; exome sequencing; in vivo; insight; mutant; neuron development; novel; particle; promoter; protein function; protein protein interaction; public health relevance; research study; transcription factor; transcriptome sequencing

 DESCRIPTION (provided by applicant): Intellectual disability is characterized by below-average cognitive ability leading to significant deficits in functioning and adaptive behaviors. Th majority of the unexplained intellectual disability cases are caused by de novo gene mutations. Four individuals with intellectual disabilities were found to have heterozygous de novo point mutations within the DNA binding domain of the transcription factor Deformed Epidermal Autoregulatory Factor-1 (DEAF1), and the resultant amino acid substitutions were shown to negatively affect DEAF1 protein function. Four additional DEAF1 variants have recently been identified from a de-identified group of samples from individuals with intellectual disabilities an autism spectrum disorders. These mutations also occur within the DNA binding domain, but the effects of these mutations on DEAF1 function are currently uncharacterized. Since DEAF1 haploin sufficiency has not been reported to result in intellectual disabilities, this suggests tha the mutant DEAF1 proteins likely exert dominant negative influences on wildtype DEAF1 function. The underlying mechanisms by which mutant DEAF1 proteins contribute to the pathogenesis producing the intellectual disabilities remain unclear. The hypothesis for this proposal is that mutations in the DEAF1 gene that diminish DEAF1 protein function cause altered neuronal activity and measurable changes in target gene expression. Specific Aim 1 will determine the effects of newly identified DEAF1 mutations on DEAF1 protein function and also assess dominant negative activity of the DEAF1 mutant proteins using both in vitro and ex vivo approaches. Specific Aim 2 will evaluate the effects of diminished DEAF1 function on neuronal activity/plasticity and target gene expression. Mice with conditional knockout of Deaf1 in the brain show anxiety-like behavior and impaired contextual memory in fear-conditioning tests and will be used to establish a role of DEAF1 in regulating neuronal activity. DEAF1 gene targets related to neuronal and cognitive function that are differentially expressed in the hippocampus of these mice will be assessed. The proposed studies will describe the functional effects of identified human de novo DEAF1 mutations and the consequences of reduced DEAF1 activity in the central nervous system. We expect that the results of these studies will also provide evidence into the causal relationship between dysregulated DEAF1 function and intellectual disability.

 描述（由申请人提供）：智力残疾的特点是低于平均水平的认知能力，导致功能和适应行为的显着缺陷。大多数原因不明的智力残疾病例是由新生基因突变引起的。发现四名智力障碍患者在转录因子变形表皮自动调节因子-1（DEAF 1）的DNA结合结构域内存在杂合从头点突变，由此产生的氨基酸取代对DEAF 1蛋白功能产生负面影响。最近，从一组来自智力残疾和自闭症谱系障碍个体的去识别样本中确定了另外四种DEAF 1变体。这些突变也发生在DNA结合结构域内，但这些突变对DEAF 1功能的影响目前尚不清楚。由于没有报道DEAF 1单倍蛋白充足导致智力残疾，这表明突变的DEAF 1蛋白可能对野生型DEAF 1功能产生显性负面影响。突变型DEAF 1蛋白参与智力残疾发病机制的机制尚不清楚。该提议的假设是，减少DEAF 1蛋白功能的DEAF 1基因突变导致神经元活性改变和靶基因表达的可测量变化。具体目标1将确定新鉴定的DEAF 1突变对DEAF 1蛋白功能的影响，并使用体外和离体方法评估DEAF 1突变蛋白的显性负活性。具体目标2将评价DEAF 1功能减弱对神经元活性/可塑性和靶基因表达的影响。大脑中有条件敲除Deaf 1的小鼠在恐惧条件反射测试中显示出焦虑样行为和受损的背景记忆，并将用于确定Deaf 1在调节神经元活动中的作用。将评估在这些小鼠海马中差异表达的与神经元和认知功能相关的DEAF 1基因靶标。拟议的研究将描述已确定的人类新发DEAF 1突变的功能效应以及中枢神经系统中DEAF 1活性降低的后果。我们期望这些研究的结果也将为DEAF 1功能失调和智力残疾之间的因果关系提供证据。