Stress Regulation of Non-Coding RNAs in Prefrontal Cortex

描述(申请人提供)：内侧前额叶皮质是压力和情绪的主要大脑媒介。在人类和动物模型中，内侧前额叶皮质功能障碍与情绪障碍、受损的恐惧消退和低效终止生理应激反应有关。内侧前额叶皮质功能障碍与许多精神疾病有关，最突出的是抑郁症和创伤后应激障碍，这些疾病由生活压力引发，并导致长期的不当应激反应。值得注意的是，前额叶皮质的基因表达对应激非常敏感，绝大多数受调控的mRNAs显示出明显的下调。最近的研究表明，非编码RNA，包括microRNAs和选择性表达的3‘-未翻译(3’-UTR)mRNA序列，在包括脑在内的许多组织中起着重要的mRNA下调作用。这一探索性方案旨在利用新开发的深度测序技术对前额叶皮质非编码RNA(MicroRNAs)和3‘UTRs(MRNAs)进行详细分析，对大鼠慢性不可预测应激对miRNA和3-UTRs的调控进行了前所未有的评估。这种不可预测的应激疗法可靠地模拟了抑郁症的生理和行为症状，允许将临床前的发现推断为人类皮质功能失调的可能机制。目的1将使用深度测序方法对C57BL6小鼠前额叶皮质中现有的以及新的慢性应激调节的miRNAs进行全面和定量的分析。目的2将深度测序方法应用于慢性应激调节3‘-非编码区序列的鉴定。在这两个目标中，后续研究将验证前额叶皮质中靶向miRNAs和3‘-UTR序列的特定调控，并使用解剖学方法将表达定位于不同的皮质亚区和细胞类型。在小鼠中发现新的应激调节的miRNAs和3‘-UTRs将有助于我们理解人类应激相关疾病的机制，并为疾病过程的干预提供可能的靶点。 公共卫生相关性：这项建议使用深度测序技术对小鼠前额叶皮质中应激调节的非编码RNA(即microRNAs)和3‘UTRs(MRNAs)的调控进行了探索性评估，以深入了解与皮质功能障碍(如抑郁症)相关的应激相关疾病的新机制。在这些研究的结论中，我们预计将开发一个迄今为止史无前例的应激调节非编码RNA调控数据库，该数据库可以被挖掘以识别在面对长期应激时危及皮质完整性的新过程。新的应激调节miRNAs和3‘-UTRs的发现将有助于我们理解人类应激相关疾病的潜在机制，并为皮质介导的精神疾病的干预开辟新的途径。

DESCRIPTION (provided by applicant): The medial prefrontal cortex is a primary brain mediator of stress and mood. In humans and in animal models, medial prefrontal cortical dysfunction is associated with emotional disturbances, impaired fear extinction and inefficient termination of physiological stress responses. Medial prefrontal cortex dysfunction is linked to numerous mental illnesses, the most prominent being depression and post-traumatic stress disorder, diseases that are triggered by life stress and result in long-term inappropriate stress responding. Notably, gene expression in the prefrontal cortex is exquisitely sensitive to stress exposure, with the vast majority of regulated mRNAs showing pronounced down-regulation. Recent studies have convincingly demonstrated that non-coding RNAs, including microRNAs and alternatively expressed 3'-unstranslated (3'-UTR) mRNA sequences, play a major role in mRNA down-regulation in numerous tissues, including brain. This Exploratory Proposal is designed to perform detailed analysis of prefrontal cortical non-coding RNAs (microRNAs) and 3' UTRs (mRNAs) using newly-developed deep sequencing technology, affording a heretofore unprecedented assessment of miRNA and 3-UTR regulation by chronic unpredictable stress in rat. The unpredictable stress regimen reliably models physiological and behavioral symptoms of depression, allowing for extrapolation of preclinical findings to putative mechanisms of functional dysregulation in human cortex. Aim 1 will use deep sequencing methods to provide a comprehensive and quantitative analysis of existing as well as novel chronic stress-regulated miRNAs in the prefrontal cortex of C57BL6 mice. Aim 2 will apply the deep sequencing methods to identification of chronic stress-regulated 3'-UTR sequences. In both Aims, follow-up studies will verify specific regulation of targeted miRNAs and 3'-UTR sequences in the prefrontal cortex, and use anatomical methods to localize expression to distinct cortical subregions and cell types. Identification of novel stress-regulated miRNAs and 3'-UTRs in mouse will inform our understanding of mechanisms underlying human stress-related disease, and provide possible future targets for intervention in disease processes. PUBLIC HEALTH RELEVANCE: This proposal provides an exploratory assessment of regulation of stress-regulated non-coding RNAs (i.e. microRNAs) and 3' UTRs (mRNAs) in the mouse prefrontal cortex using deep sequencing technologies, to provide insight into novel mechanisms of stress-related diseases associated with cortical dysfunction (e.g., depression). At the conclusion of these studies, we anticipate development of a heretofore unprecedented database of stress-regulated non-coding RNA regulation that can be mined to identify novel processes endangering cortical integrity in the face of prolonged stress. Identification of novel stress-regulated miRNAs and 3'-UTRs will inform our understanding of mechanisms underlying human stress-related disease, and define new avenues for intervention in cortically-mediated mental illness.