Prenatal alcohol exposure generates vulnerability to the proinflammatory effects of morphine and adverse neuroimmune consequences

项目摘要 妊娠期接触酒精会导致一系列轻度到重度的认知和行为障碍， 胎儿酒精谱系障碍（FASD），在美国的患病率高达~ 4.8%。 尽管关于产前酒精暴露（PAE）长期后果的临床研究很少，但高 物质使用的普遍程度（例如，酒精，阿片类药物）在成人FASD人群中观察到， 酒精或阿片类药物引起的异常神经免疫相互作用。该提案审查了 阿片类镇痛药物治疗后PAE是否诱导动物神经免疫功能障碍的易感性 模型在PAE动物模型中，促炎细胞因子（包括白细胞介素-1 β）的产生增加， (IL-1β）在成人中枢神经系统（CNS）中的分布。我们最近的动物模型研究表明， PAE是发生CNS病理学的风险因素，如病理性疼痛（异常性疼痛），通过以下作用 通过释放IL-1β和肿瘤坏死因子-α（TNF-α）激活脊髓星形胶质细胞和巨噬细胞。一 导致IL-1β和TNF-α增加的关键先天性免疫受体是Toll样受体（TLR 4）， 其活化产生下游NLRP 3炎性体信号传导和随后的IL-1β。奇怪的是， 吗啡是一种作用于μ-阿片受体的标准疼痛治疗剂，也能激活促炎性TLR 4 信号传导导致对立过程; μ-阿片样物质细胞作用，以减弱疼痛和免疫TLR 4细胞 造成痛苦的行为。因此，可能需要更高剂量的吗啡来实现疼痛缓解。 一些人口。基于这一背景，我们推测PAE可能产生生物脆弱性， CNS病理学通过致敏TLR 4对内源性因子和吗啡的反应， TLR4。因此，这项建议的一个目标是确定小鼠吗啡治疗是否揭示了潜在的影响， PAE对神经免疫功能的影响，导致慢性CNS病理后果。初步数据显示 吗啡暴露矛盾地延长了轻微神经损伤的成年PAE小鼠的异常性疼痛过程。的 PAE和阿片样物质诱导的TLR 4作用的潜在趋同性完全未被探索。所以我们的 一种假设是：PAE启动了TLR 4-NLRP 3-IL-1β通路，该通路被内源性疼痛增强 因子和吗啡与TLR 4相互作用导致慢性病理性神经免疫功能。目标 （I）PAE是否增强吗啡介导的TLR 4和/或NLRP 3活化， （II）PAE增敏吗啡对TLR 4/NLRP 3的作用 脊髓星形胶质细胞和外周巨噬细胞的反应，以及（III）非编码circ-Vopp 1的贡献 在响应于吗啡的PAE引发的TLR 4信号传导中的失调。研究结果将提供新的证据， PAE是阿片类药物不良反应的风险因素，并将确定新的靶点以缓解 FASD个体中不良TLR 4作用导致的神经免疫功能障碍。

PROJECT SUMMARY Exposure to alcohol during gestation can lead to a constellation of mild to severe cognitive and behavioral deficits referred to as Fetal Alcohol Spectrum Disorders (FASD), having a US prevalence as high as ~4.8%. Although clinical studies on long-term consequences of prenatal alcohol exposure (PAE) are sparse, high prevalence of substance use (e.g., alcohol, opioids) is observed in adult FASD populations which may stem from aberrant neuroimmune interactions following exposure to alcohol or opioids. This proposal examines whether PAE induces susceptibility to neuroimmune dysfunction following opioid pain therapy in animal models. In PAE animal models, heightened production of proinflammatory cytokines including interleukin-1β (IL-1β) in the adult central nervous system (CNS) is observed. Our recent work in animal models suggest that PAE is a risk factor for developing CNS pathology such as pathological pain (allodynia) through the actions of spinal astrocyte and macrophage activation by their release of IL-1β and tumor necrosis factor-α (TNF-α). A critical innate immune receptor underlying increased IL-1β and TNF-α is Toll Like Receptor (TLR4), that upon its activation, produces downstream NLRP3 inflammasome signaling and consequent IL-1β. Curiously, morphine, a standard pain therapeutic acting on the μ-opioid receptor, also activates proinflammatory TLR4 signaling resulting in opponent processes; µ-opioid cellular actions to blunt pain and immune TLR4 cellular actions that cause pain. Consequently, higher doses of morphine may be required to achieve pain alleviation in some populations. Based on this background, we speculate that PAE may generate biological vulnerability to CNS pathology by sensitizing TLR4 responses to endogenous factors and morphine that readily interact with TLR4. Thus, one goal of this proposal is to identify if morphine treatment in mice unmasks the insidious effects of PAE on neuroimmune function resulting in chronic CNS pathological consequences. Preliminary data show morphine exposure paradoxically prolongs the course of allodynia in minor nerve-injured adult PAE mice. The potential convergence of PAE and opioid-induced TLR4 actions is entirely unexplored. Therefore, our hypothesis is: PAE primes the TLR4-NLRP3-IL-1β pathway that is unmasked by endogenous pain-enhancing factors and morphine interacting with TLR4 resulting in chronic pathological neuroimmune function. The Aims of the proposal will determine if : (I) PAE augments morphine-mediated TLR4 &/or NLRP3 activation in minor nerve-injured mice causing prolonged allodynia, (II) PAE sensitizes the effects of morphine on TLR4/NLRP3 responses in spinal astrocytes and peripheral macrophages, and (III) the contribution of non-coding circ-Vopp1 dysregulation in PAE-primed TLR4 signaling in response to morphine. Results will provide new evidence for PAE as a risk factor for adverse opioid responses and will identify novel targets to mitigate the risk of neuroimmune dysfunction from adverse TLR4 actions in individuals with FASD.