TAFopathies Result from Derangements in Transcriptional Control of Metabolism

TAF 病是由代谢转录控制紊乱引起的

• 批准号: 10579824
• 负责人: Jamison Leid
• 金额: $ 3.27万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10579824
• 关键词: 3-Dimensional; Adaptive Behaviors; Affect; Anatomy; Basal metabolic rate; Binding Proteins; Biological Assay; Bromodomain; CRISPR/Cas technology; Cardiac; Cardiac development; Cell Differentiation process; Cell Respiration; ChIP-seq; Clinical; Complex; Craniofacial Abnormalities; Data; Data Analyses; Defect; Development; Disease; Embryo; Embryonic Development; Enhancers; Exhibits; Experimental Designs; Experimental Models; Fertilization; Gene Expression; General Population; Genes; Genetic; Genetic Enhancer Element; Genetic Screening; Genetic Transcription; Goals; Heart; Heart Abnormalities; Heart failure; Hi-C; Histology; Hour; Human; In Situ Hybridization; Individual; Intellectual functioning disability; Interruption; Lead; Learning; Link; Mammals; Mediating; Metabolic; Metabolic Activation; Metabolic dysfunction; Metabolism; Mitochondria; Modeling; Mutation; Neurodevelopmental Disorder; Neurologic; Nonsense Mutation; Pathogenicity; Patients; Pattern; Phenotype; Proteins; RNA; RNA Polymerase II; Regulation; Respiration; Role; Signal Transduction; Site; Structural defect; Syndrome; TAF1 gene; TAF2 gene; TAF5 gene; TAF6 gene; TATA-Binding Protein Associated Factors; TATA-Box Binding Protein; Techniques; Testing; Time; Transcriptional Activation; Transcriptional Regulation; United States; Variant; Ventricular; X ray microscopy; Zebrafish; chromatin immunoprecipitation; cognitive function; computational pipelines; congenital heart disorder; craniofacial; craniofacial development; experimental study; innovation; loss of function mutation; mutant; neurodevelopment; novel; patient subsets; programs; promoter; recruit; skills; spatiotemporal; transcription factor; transcriptome sequencing; treatment strategy

Project Summary Intellectual disability is a disabling neurodevelopmental disorder that affects 2-3% of the general population. Often times, these intellectual disabilities are accompanied by additional developmental abnormalities. These syndromic forms of intellectual disability frequently have a genetic basis. Recent studies have identified a subset of intellectual disability patients with pathogenic variants in key components of the TATA-binding protein associated factors (TAFs). These patients also have craniofacial defects and congenital heart disease. This syndrome has been termed as a TAFopathy and includes mutations in TATA binding protein (TBP), TAF1, TAF2, and TAF6. Through a forward genetic screen, we have recovered a lethal nonsense mutation in TAF5. Mutant embryos have craniofacial hypoplasia, ventricular hypoplasia, and heart failure at 96 hours post fertilization. CRISPR/Cas9 mediated gene editing revealed that this phenotype was recapitulated in TAF1 KO and TAF5 KO embryos. TAF5 KOs show significant metabolic dysfunction that may precede the anatomical defects observed at 96 hours post fertilization. Together, these findings support a regulatory role for TAFs in coordinating transcriptional activation of metabolic programming during embryogenesis. The TAFs form the general transcription factor TFIID, which recruits RNA Polymerase II to form the preinitation complex at sites of transcription. Studies have shown that individual TAFs are not necessary for general transcription. Other studies have identified functional domains of TAF1 that activate transcription through interactions with TBP and enhancer elements that are sufficient to recruit RNA Polymerase II to sites of transcription. In conjunction with our data showing metabolic dysfunction, we hypothesize that TAFs regulate metabolic programs by linking enhancers and promoters of metabolic genes through TFIID assembly. Our current efforts are focused towards completing our phenotypic characterization of TAF1 KOs and TAF5 KOs with respect to craniofacial- and neuro-development. We plan to use innovative and cutting-edge techniques, including x-ray microscopy, RNA- Scope in situ hybridization, mitochondrial respiration assays, ChIP-seq, and Hi-C to achieve this goal. The data obtained will help us elucidate basic disease mechanisms of TAFopathy. These findings would also indicate a secondary function for general transcription factors to regulate metabolic programming during embryogenesis. .

项目摘要 智力障碍是一种致残性神经发育障碍，影响2-3%的普通人群。 通常情况下，这些智力残疾伴随着额外的发育异常。这些 综合症形式的智力残疾往往具有遗传基础。最近的研究发现， TATA结合蛋白关键组分中存在致病性变异的智力残疾患者亚组 相关因素（TAFs）。这些患者也有颅面缺陷和先天性心脏病。这 综合征被称为TAF病，包括TATA结合蛋白（TBP），TAF 1， TAF 2和TAF 6。通过正向遗传筛选，我们在TAF 5中发现了一个致命的无义突变。 突变胚胎在96小时后出现颅面发育不全、心室发育不全和心力衰竭。 受精CRISPR/Cas9介导的基因编辑揭示了这种表型在TAF 1 KO中重现。 和TAF 5 KO胚胎。TAF 5科斯表现出显著的代谢功能障碍，其可能先于解剖结构的改变。 在受精后96小时观察到缺陷。总之，这些发现支持TAF在以下方面的调节作用： 协调胚胎发生期间代谢编程的转录激活。TAF形成了 一般转录因子TFIID，其募集RNA聚合酶II以在以下位点形成预起始复合物： 转录。研究表明，单个TAF对于一般转录是不必要的。其他 研究已经鉴定了TAF 1的功能结构域，其通过与TBP的相互作用激活转录， 增强子元件足以将RNA聚合酶II募集到转录位点。结合 我们的数据显示代谢功能障碍，我们假设TAFs通过连接调节代谢程序， 通过TFIID组装的代谢基因的增强子和启动子。我们目前的工作重点是 完成我们对TAF 1科斯和TAF 5科斯在颅面-和 神经发育我们计划使用创新和尖端技术，包括X射线显微镜，RNA- 范围包括原位杂交、线粒体呼吸测定、ChIP-seq和Hi-C以实现这一目标。数据 这些结果将有助于我们阐明TAF病的基本发病机制。这些发现还表明， 一般转录因子在胚胎发生过程中调节代谢程序的次要功能。 .