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大脑调节器。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无效终止有关。内侧前额皮质功能障碍与许多精神疾病有关，其中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并导致长期不适当的压力反应。值得注意的是，前额皮质中的基因表达对压力暴露极其敏感，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证明，非编码 RNA，包括 microRNA 和替代表达的 3'-非翻译 (3'-UTR) mRNA 序列，在包括大脑在内的许多组织中的 mRNA 下调中发挥着重要作用。该探索性提案旨在利用新开发的深度测序技术对前额皮质非编码 RNA (microRNA) 和 3' UTR (mRNA) 进行详细分析，从而对大鼠慢性不可预测应激的 miRNA 和 3-UTR 调节提供前所未有的评估。不可预测的压力疗法可靠地模拟了抑郁症的生理和行为症状，从而可以将临床前发现外推到人类皮质功能失调的推定机制。目标 1 将使用深度测序方法对 C57BL6 小鼠前额皮质中现有的以及新型的慢性应激调节 miRNA 进行全面的定量分析。目标 2 将应用深度测序方法来鉴定慢性应激调节的 3'-UTR 序列。在这两个目标中，后续研究将验证前额皮质中目标 miRNA 和 3'-UTR 序列的特异性调节，并使用解剖学方法将表达定位到不同的皮质亚区域和细胞类型。在小鼠中鉴定新型应激调节 miRNA 和 3'-UTR 将有助于我们了解人类应激相关疾病的机制，并为干预疾病过程提供可能的未来目标。 公共健康相关性：该提案利用深度测序技术对小鼠前额皮质中压力调节非编码 RNA（即 microRNA）和 3' UTR（mRNA）的调节进行了探索性评估，以深入了解与皮质功能障碍（例如抑郁症）相关的压力相关疾病的新机制。在这些研究的结论中，我们预计将开发出迄今为止前所未有的压力调节非编码 RNA 调节数据库，可以挖掘该数据库来识别在长期压力下危及皮质完整性的新过程。新型压力调节 miRNA 和 3'-UTR 的鉴定将有助于我们了解人类压力相关疾病的机制，并确定干预皮质介导的精神疾病的新途径。

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.