Antisense RNA Mediated Epigenetic Regulation of Brain Derived Neurotrophic Factor

描述（由申请人提供）：低丰度非编码反义rna是从许多基因组位点的相反链转录而来。这些内源性调控元件被证明调节相应的mRNA表达，在某些情况下，通过招募表观遗传酶诱导位点特异性染色质修饰。抑制或去除这一反义转录子亚群可导致感觉mRNA和功能蛋白的位点特异性上调。我们发现脑源性神经营养因子（BDNF）受反义rna的调控，反义rna通过转录抑制感觉mRNA和蛋白质的表达。本提案的主要目的是研究非编码rna作为BDNF表达的表观遗传调节剂的作用。首先，我们将在体外研究反义rna介导的BDNF调控机制，在人和小鼠神经上皮细胞的原代培养中。然后，我们将开发工具来定量测量BDNF反义RNA （BDNF- AS）与表观遗传酶EZH2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，我们将利用Rett综合征小鼠模型（Mecp2缺失小鼠）研究Bdnf- as敲低对Bdnf位点的体内影响。根据该提案计划的研究将阐明非编码反义rna，特别是BDNF- as，调节BDNF位点表达的分子机制。我们独特的方法将导致鉴定非编码RNA-蛋白质相互作用的分子基础。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。

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.