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

项目总结 怀孕期间接触酒精会导致一系列从轻度到重度的认知和行为 缺乏症被称为胎儿酒精谱系障碍(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，但数量很多 在成人FASD人群中观察到使用物质(如酒精、阿片类药物)的流行率，这可能会导致 暴露在酒精或阿片类药物后的异常神经免疫相互作用。这份提案审查了 PAE是否导致阿片类疼痛治疗后动物神经免疫功能障碍的易感性 模特们。在PAE动物模型中，包括白细胞介素1β在内的促炎细胞因子的产生增加 本文观察了成人中枢神经系统中IL-1、β的表达。我们最近在动物模型上的研究表明 PAE是发生中枢神经系统病理的危险因素，如通过以下作用发生病理性疼痛(痛觉异常) 脊髓星形胶质细胞和巨噬细胞通过释放IL-1β和肿瘤坏死因子α(肿瘤坏死因子-α)来激活。一个 IL-1β和肿瘤坏死因子-α升高背后的关键先天免疫受体是Toll样受体，在 它的激活，产生下游的NLRP3炎症体信号和随后的IL-1β。奇怪的是， 吗啡是作用于μ-阿片受体的标准止痛药，也能激活促炎因子TLR4 信号导致相反的过程；µ-阿片类细胞作用于钝化疼痛和免疫TLR4细胞 导致痛苦的行为。因此，可能需要更高剂量的吗啡来缓解疼痛。 一些种群。基于这一背景，我们推测PAE可能会对 通过敏化TLR4对内源性因素和容易与之相互作用的吗啡的反应来实现中枢神经系统病理 TLR4.因此，这项提议的一个目标是确定吗啡在小鼠身上的治疗是否暴露了潜在的影响 PAE对神经免疫功能的影响导致慢性中枢神经系统病理后果。初步数据显示 在轻微神经损伤的成年PAE小鼠中，吗啡暴露矛盾地延长了痛觉过敏的病程。这个 PAE和阿片类药物诱导的TLR4作用的潜在趋同性完全未被探索。因此，我们的 假说是：PAE启动了TLR4-NLRP3-IL-1β通路，该通路未被内源性疼痛增强所掩盖 因子和吗啡与TLR4相互作用导致慢性病理性神经免疫功能。目标 将决定：(I)PAE是否增强吗啡介导的TLR4和/或NLRP3在未成年人中的激活 神经损伤致小鼠长时痛觉异常(II)PAE增敏吗啡对TLR4/NLRP3的影响 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-Vopp1的作用 PAE诱导的TLR4信号对吗啡反应的失调。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，并将确定新的目标，以减轻 FASD患者TLR4不良反应所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.