Molecular and Behavioral Neurobiology of Transcription Factor TCF4

转录因子 TCF4 的分子和行为神经生物学

• 批准号: 8883724
• 负责人: John David Sweatt
• 金额: $ 36.75万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8883724
• 关键词: Accounting; Acetylesterase; Acute; Address; Adult; Alleles; Anatomy; Area; Auditory; Autistic Disorder; BHLH Protein; Behavior; Behavioral; Binding; Bioinformatics; Boxing; Brain; Breeding; Categories; Chromatin Structure; Cognition; Cognitive; Complex; Coupled; DNA; DNA Methylation; Dendritic Spines; Development; Diagnosis; Epigenetic Process; Etiology; Exhibits; Exons; Family; Gene Targeting; Genes; Genetic; Genetic Engineering; Genetic Transcription; Genetically Engineered Mouse; Genomics; Health; Hippocampus (Brain); Histone Deacetylase; Histone Deacetylase Inhibitor; Histones; Homo; Human; Impaired cognition; Introns; Knock-out; Laboratories; Language; Language Development; Learning; Long-Term Potentiation; Mediating; Memory; Memory impairment; Messenger RNA; Modeling; Molecular; Molecular Biology; Motor; Mus; Mutation; Nervous System Physiology; Neuraxis; Neurobiology; Neurons; Patients; Pharmaceutical Preparations; Pharmacotherapy; Physiological; Predisposition; Prosencephalon; Proteins; Regulation; Regulatory Element; Resources; Role; Route; Schizophrenia; Small RNA; Social Behavior; Social Interaction; Susceptibility Gene; Synapses; Synaptic plasticity; Syndrome; TCF7L2 gene; Testing; Transcription factor genes; Transcriptional Activation; Transcriptional Regulation; Veins; Virus; attenuation; autistic behaviour; base; calmodulin-dependent protein kinase II; cognitive function; epigenomics; experience; gene function; genetic risk factor; genome wide association study; improved; in vivo; inhibitor/antagonist; innovation; learned behavior; loss of function; mouse model; next generation sequencing; novel; promoter; response; social communication; synaptic function; transcription factor; transcriptome sequencing; transcriptomics

DESCRIPTION (provided by applicant): The basic helix-loop-helix transcription factor TCF4 (aka E2-2) has been implicated broadly in human CNS function. TCF4 was confirmed in several large GWAS studies as one of the rare highly replicated schizophrenia susceptibility genes, and when haplo-insufficient TCF4 is the causative factor for Pitt-Hopkins Syndrome (PTHS). PTHS has compelling attributes in terms of human cognitive function, being associated with pronounced memory deficits, autistic behaviors, and importantly an almost complete lack of language development. Thus, understanding the roles and function of TCF4 in the CNS is highly significant with respect to human language and auditory cognition, memory function, autism spectrum behavior, and schizophrenia susceptibility. Despite the clear importance of TCF4 function in the human CNS, the basic neurobiology of TCF4 has been only sparsely studied. Thus, this Project will use genetically engineered mouse models to assess the role of TCF4 in memory, social interactions and communication, hippocampal synaptic plasticity, synaptic anatomy, and epigenomic and transcriptional regulation in the CNS. The central hypothesis of the proposed studies is that TCF4 regulates the brain's ability to trigger long-term synaptic plasticity and memory formation by actively regulating transcriptional activity in response to behavioral experience. The approach will be to use both germline constitutive knockout and acute knockdown or deletion of TCF4 in the adult mouse hippocampus, coupled with extensive behavioral, electrophysiological, and transcriptomic/epigenomic characterization, to determine if TCF4 function is necessary in an ongoing fashion for normal memory and synaptic plasticity in the mature CNS. The studies will capitalize not only upon the available germline knockout line but also upon an already-available floxed TCF4 allele mouse line, and combine these mice with both virus- driven cre expression in the adult hippocampus and post-developmental CaMKII promoter-driven forebrain neuron-selective cre expression to achieve inducible post-developmental attenuation of CNS TCF4 function in vivo. Using these novel mouse models we will undertake three Specific Aims. Aim 1 will test the hypothesis that TCF4-deficient mice exhibit cognitive memory and social interaction deficits, and also test the hypothesis that TCF4-deficient mice exhibit altered synaptic plasticity. Aim 2 will comprise a significant translational component of the studies and will test the hypothesis that transcription-promoting Histone DeAcetylase Inhibitors (HDACi) will restore learning, memory, and synaptic plasticity in TCF4-deficient mice, as proof-of- concept that behavioral deficits and plasticity deficits in PTHS model mice can be reversed pharmacologically with HDACi. Aim 3 will test the hypothesis that loss of TCF4 function triggers secondary alterations in chromatin structure, DNA methylation, and gene transcription in the mature CNS. Toward this end, we will use the unbiased approach of utilizing MBD-seq, mRNA-seq, and small RNA-seq approaches for a comprehensive analysis of potential transcriptomic and epigenomic alterations in TCF4-deficient mice.

描述（由申请人提供）：基本螺旋-环-螺旋转录因子TCF4（又名E2-2）已广泛涉及人类中枢神经系统功能。 TCF4在几项大型GWAS研究中被证实是罕见的高度复制的精神分裂症易感基因之一，并且单倍体不足的TCF4是皮特-霍普金斯综合征（PTHS）的致病因素。 PTHS 在人类认知功能方面具有引人注目的属性，与明显的记忆缺陷、自闭症行为以及重要的是几乎完全缺乏语言发展有关。因此，了解 TCF4 在 CNS 中的作用和功能对于人类语言和听觉认知、记忆功能、自闭症谱系行为和精神分裂症易感性具有非常重要的意义。尽管 TCF4 功能在人类 CNS 中的重要性显而易见，但 TCF4 的基本神经生物学研究却很少。因此，该项目将使用基因工程小鼠模型来评估TCF4在记忆、社交互动和沟通、海马突触可塑性、突触解剖学以及中枢神经系统表观基因组和转录调控中的作用。拟议研究的中心假设是，TCF4 通过主动调节转录活动来响应行为体验，从而调节大脑触发长期突触可塑性和记忆形成的能力。该方法将使用成年小鼠海马体中TCF4的种系组成型敲除和急性敲除或删除，再加上广泛的行为、电生理学和转录组/表观基因组特征，以确定TCF4功能对于成熟中枢神经系统中的正常记忆和突触可塑性是否是持续必需的。这些研究不仅利用现有的种系敲除系，而且利用已经可用的 floxed TCF4 等位基因小鼠系，并将这些小鼠与成年海马中病毒驱动的 cre 表达和发育后 CaMKII 启动子驱动的前脑神经元选择性 cre 表达相结合，以实现体内 CNS TCF4 功能的诱导性发育后衰减。使用这些新颖的小鼠模型，我们将实现三个具体目标。目标 1 将检验 TCF4 缺陷小鼠表现出认知记忆和社交互动缺陷的假设，并检验 TCF4 缺陷小鼠表现出突触可塑性改变的假设。目标 2 将包括重大转化 研究的组成部分，并将测试转录促进组蛋白去乙酰化酶抑制剂 (HDACi) 将恢复 TCF4 缺陷小鼠的学习、记忆和突触可塑性的假设，作为 PTHS 模型中行为缺陷和可塑性缺陷的概念证明 HDACi 可以在药理学上逆转小鼠。目标 3 将检验以下假设：TCF4 功能的丧失会引发成熟 CNS 中染色质结构、DNA 甲基化和基因转录的二次改变。为此，我们将使用 MBD-seq、mRNA-seq 和小 RNA-seq 方法来全面分析 TCF4 缺陷小鼠中潜在的转录组和表观基因组改变。