Antisense RNA Mediated Epigenetic Regulation of Brain Derived Neurotrophic Factor

反义RNA介导的脑源性神经营养因子的表观遗传调控

• 批准号: 8792878
• 负责人: Mohammad Ali Faghihi
• 金额: $ 33.47万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8792878
• 关键词: Antisense RNA; Behavioral; Beryllium; Binding; Biological Assay; Biotechnology; Brain; Brain-Derived Neurotrophic Factor; Cells; Chromatin; Chromatin Structure; Chromosomes; Complementary DNA; Complex; Data; Dependovirus; Disease; Enhancers; Enzymes; Epigenetic Process; Excision; Future; Gene Expression; Gene Expression Regulation; Genetic Transcription; Genomics; Goals; Health; Human; Immunoprecipitation; In Vitro; Individual; Knockout Mice; Lead; Length; Light; Long-Term Effects; Measures; Mediating; Messenger RNA; Molecular; Molecular Conformation; Mus; Nature; Neonatal; Neuroepithelial Cells; Neurons; Oligonucleotides; Pattern; Proteins; Publishing; RNA; RNA Splicing; RNA purification; RNA-Protein Interaction; Regulation; Regulatory Element; Research Proposals; Rett Syndrome; Reverse Transcriptase Polymerase Chain Reaction; Role; Specificity; Structure; Technology; Testing; Time; Tissues; Transcript; Transcription Initiation Site; Untranslated RNA; Up-Regulation; Variant; behavior test; cell type; chromatin immunoprecipitation; chromatin modification; chromatin remodeling; epigenetic regulation; histone methyltransferase; in vivo; insight; loss of function; mRNA Expression; mouse model; neuropsychological; neurotrophic factor; novel; osmotic minipump; research study; small hairpin RNA; therapeutic target; tool; treatment duration

DESCRIPTION (provided by applicant): Low abundance noncoding antisense RNAs are transcribed from the opposite strand of many genomic loci. These endogenous regulatory elements are shown to regulate corresponding mRNA expression, in some cases through the recruitment of epigenetic enzymes that induce locus-specific chromatin modifications. Inhibition or removal of this subset of antisense transcripts leads to locus-specific up-regulation of sense mRNA and functional protein. We found that brain-derived neurotrophic factor (BDNF) is under the regulatory control of antisense RNAs that transcriptionally suppress the expression of sense mRNA and protein. The main goal of this proposal is to study the role of noncoding RNAs as epigenetic modulators of BDNF expression. Initially, we will study the mechanism of antisense RNA-mediated regulation of BDNF, in vitro, in primary cultures of human and mouse neuroepithelial cells. We will then develop tools to quantitatively measure the interaction between BDNF antisense RNA (BDNF- AS) and the epigenetic enzyme EZH2 to investigate the structural and/or sequence requirements of this interaction. Lastly, we will examine the in vivo effects of Bdnf-AS knockdown on the Bdnf locus, utilizing a mouse model of Rett syndrome (Mecp2 null mice). Studies planned under this proposal will shed light on the molecular mechanisms by which noncoding antisense RNAs, in particular BDNF-AS, regulate expression at the BDNF locus. Our unique approach will lead to identification of the molecular underpinning of noncoding RNA- protein interactions. Finally, we will study the in vivo regulatory role of endogenous BDNF-AS in a mouse model of Rett syndrome. Overall these projects will pave the road for future epigenetic studies on other genomic loci, serving to establish novel noncoding RNA therapeutic targets for several neuropsychological disorders.

描述（由申请人提供）：低丰度非编码反义RNA从许多基因组基因座的相对链转录。这些内源性调控元件显示出调节相应的mRNA表达，在某些情况下通过募集诱导基因座特异性染色质修饰的表观遗传酶。抑制或去除该反义转录物子集导致有义mRNA和功能蛋白的基因座特异性上调。我们发现，脑源性神经营养因子（BDNF）是在调控下的反义RNA，转录抑制正义mRNA和蛋白质的表达。本研究的主要目的是研究非编码RNA作为BDNF表达的表观遗传调节因子的作用。首先，我们将在体外研究反义RNA介导的BDNF调节机制，在人类和小鼠神经上皮细胞的原代培养。然后，我们将开发工具来定量测量BDNF反义RNA（BDNF- AS）和表观遗传酶EZH 2之间的相互作用，以研究这种相互作用的结构和/或序列要求。最后，我们将研究在体内的影响，Bdnf-AS敲低的Bdnf基因座，利用小鼠模型的Rett综合征（Mecp 2空小鼠）。根据这一建议计划的研究将阐明非编码反义RNA，特别是BDNF-AS，调节BDNF基因座表达的分子机制。我们独特的方法将导致非编码RNA-蛋白质相互作用的分子基础的鉴定。最后，我们将研究内源性BDNF-AS在Rett综合征小鼠模型中的体内调节作用。总的来说，这些项目将为未来对其他基因组位点的表观遗传学研究铺平道路，为几种神经心理疾病建立新的非编码RNA治疗靶点。