Stress Regulation of Non-Coding RNAs in Prefrontal Cortex

描述（由申请人提供）：内侧前额叶皮层是压力和情绪的主要脑介质。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、恐惧消退受损和生理应激反应的低效终止有关。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，这些疾病是由生活压力引发的，导致长期不适当的压力反应。值得注意的是，前额叶皮层的基因表达对压力暴露非常敏感，绝大多数受调节的mrna表现出明显的下调。最近的研究令人信服地表明，非编码rna，包括microRNAs和替代表达的3'-未翻译（3'-UTR） mRNA序列，在包括大脑在内的许多组织中mRNA下调起主要作用。本探索性提案旨在利用新开发的深度测序技术对大鼠前额叶皮层非编码rna （microRNAs）和3′utr （mrna）进行详细分析，为慢性不可预测应激对大鼠miRNA和3′utr的调控提供迄今为止前所未有的评估。不可预测的应激方案可靠地模拟了抑郁症的生理和行为症状，允许将临床前发现推断为人类皮质功能失调的假定机制。Aim 1将使用深度测序方法对C57BL6小鼠前额皮质中现有的和新的慢性应激调节mirna进行全面定量分析。目标2将应用深度测序方法鉴定慢性应激调节的3'-UTR序列。在这两个Aims中，后续研究将验证前额叶皮层中靶向mirna和3'-UTR序列的特异性调控，并使用解剖学方法将表达定位到不同的皮层亚区和细胞类型。在小鼠中鉴定新的应激调节mirna和3'- utr将有助于我们了解人类应激相关疾病的机制，并为未来干预疾病过程提供可能的靶点。

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.