Postmortem studies of CRF-PACAP in human PTSD (Berretta)

摘要：项目 5（人类 PTSD/Berretta 中 PACAP-CRF 的死后研究） 令人信服的证据表明促肾上腺皮质激素释放因子（CRF）和垂体腺苷酸环化酶激活 多肽（PACAP）及其相互作用对应激反应做出了关键贡献， 焦虑、昼夜节律调节和创伤后应激障碍的病理生理机制 （创伤后应激障碍）。人们对所涉及的潜在神经回路知之甚少，但重要线索指向床 终纹核 (BNST)、杏仁核 (AMG)、背侧前扣带回 (dACG) 和 下丘脑（HPTh）是这些功能的关键调节者。目前的证据表明 CRF 和 PACAP 导致 PTSD 的机制可能具有性别特异性，这增加了潜在大脑的可能性 男性和女性的变化和潜在的治疗目标可能不同。当前和初步数据表明 PACAP信号传导可能直接影响CRF表达细胞，并且PACAP及其昼夜节律表达 同源受体 PAC1R 及其对 CRF 系统的后续调节可能会在整个过程中发生变化。 一天。此类变化可能会导致与 DSM 定义相关的睡眠/觉醒周期中断 疾病包括创伤后应激障碍（PTSD）、重度抑郁症和焦虑症。令人惊讶的是，几乎没有任何信息 可用于人类 AMG、BNST、dACG 和 CRF 和 PACAP 信号通路的细胞水平表达 HPTh、它们与昼夜节律的关系以及 CRF/PACAP 相互作用在 PTSD 的神经病理学。我们的首要假设是影响 CRF/PACAP 通路的异常 BNST、AMG、dACG 和 HPTh 有助于 PTSD 的病理学。在目标 1 中，我们解决了一个关键差距 关于 CRF、PACAP 及其区域、性别和昼夜节律特异性表达和分布的知识 健康人脑中的受体。我们的假设是 CRF、PACAP 和 mRNA 的蛋白质和 mRNA 表达 它们的受体具有区域和性别特异性；特别是，我们预测 PACAP 受体将表现出细胞和 CRF 阳性神经元中的性别特异性和表达，支持 PACAP 调节这些神经元的假设 神经元以性别依赖的方式。在目标 2 和 3 中，我们检查蛋白质、基因表达是否 和细胞水平——dACG、AMG、BNST 和 HPTh 的信号通路在 PTSD 中发生改变。我们的假设 与健康对照相比，患有 PTSD 的受试者的 CRF 和 PACAP 信号通路将发生改变， PACAP 表达增加与 CRF 信号通路变化相关，在区域、性别和 昼夜节律特定的方式。我们预测 PACAP 阳性细胞和轴突的增加，反映 局部 PACAP 表达增加以及来自下丘脑输入的 PACAP 表达增加，将伴随表达改变 PACAP 受体和 CRF 阳性细胞中下游信号通路的影响。项目 5 可能会识别 CRF 和 PACAP 系统和回路在与压力相关的疾病（例如 PTSD）中发生了根本性改变，并且是一种 中心的关键联系，增强了其他（临床前、临床）要素，并被其他要素所增强。

SUMMARY: PROJECT 5 (POSTMORTEM STUDIES OF PACAP-CRF IN HUMAN PTSD/BERRETTA) Compelling evidence indicates that corticotropic releasing factor (CRF) and pituitary adenylyl cyclase-activating polypeptide (PACAP), as well as their interactions together, make critical contributions to stress responses, anxiety, circadian rhythm regulation, and the pathophysiological mechanisms of post-traumatic stress disorder (PTSD). The underlying neural circuitry involved is poorly understood, but important clues point to the bed nucleus of the stria terminalis (BNST), amygdala (AMG), dorsal anterior cingulate gyrus (dACG), and the hypothalamus (HPTh) as critical regulators of these functions. Current evidence indicates that CRF and PACAP mechanisms that contribute to PTSD may be sex-specific, raising the possibility that the underlying brain changes and potential therapeutic targets may differ in males and females. Current and preliminary data suggest that PACAP signaling may directly affect CRF expressing cells and that circadian expression of PACAP and its cognate receptor PAC1R, and their subsequent regulation of CRF systems, may vary during the course of the day. Such variations may potentially contribute to disruptions of sleep/wake cycles associated with DSM-defined illnesses including PTSD, major depression, and anxiety disorders. Surprisingly, virtually no information is available on cell-level expression of CRF and PACAP signaling pathways in the human AMG, BNST, dACG and HPTh, their relationships to circadian rhythms, and the involvement of CRF/PACAP interactions in the neuropathology of PTSD. Our overarching hypothesis is that abnormalities affecting CRF/PACAP pathways in the BNST, AMG and dACG and HPTh contribute to the pathology of PTSD. In Aim 1, we address a critical gap of knowledge on the region-, sex- and circadian- specific expression and distribution of CRF, PACAP, and their receptors in healthy human brain. Our hypothesis is that protein and mRNA expression of CRF, PACAP, and their receptors are region- and sex-specific; in particular, we predict that PACAP receptors will show cell- and sex- specificity and expression in CRF-positive neurons, supporting the hypothesis that PACAP regulates these neurons in a sex dependent manner. In Aims 2 and 3, we examine whether—at the protein, gene expression, and cellular level—signaling pathways in the dACG, AMG, BNST and HPTh are altered in PTSD. Our hypothesis is that CRF and PACAP signaling pathways will be altered in subjects with PTSD, relative to healthy controls, with increased PACAP expression correlated with CRF signaling pathway changes, in a region-, sex-, and circadian rhythm-specific manner. We predict that increases of PACAP-positive cells and axons, reflecting increased PACAP expression locally and from hypothalamic inputs, will be accompanied by altered expression of PACAP receptors and down-stream signaling pathways in CRF-positive cells. Project 5 may identify CRF and PACAP systems and circuits as being fundamentally altered in stress-related illnesses such as PTSD, and is a key nexus of the Center that enhances, and is enhanced by, the other (preclinical, clinical) elements.