Prenatal Opioid Exposure and Inflammation: The Role of the Microbiome and Epigenome.

项目摘要 在持续的阿片类药物危机之后，怀孕期间阿片类药物使用的发生率增加促使了这一点 有针对性地调查产前阿片类药物暴露的影响。在这里，我们研究分子机制 通过产前阿片类药物暴露产生全身炎症和免疫抑制表型 在以后的生活中。这一结果与全身性炎症反应综合征(SIRS)一致，随后是 相应的限制有害全身的代偿性抗炎反应综合征(CARS) 免疫反应。最近的研究表明，微生物失调在全身性炎症和 巨噬细胞在CARS介导的免疫抑制中的表观遗传修饰。具体来说，成人模特 研究表明，在慢性阿片类药物暴露后，肠道微生物失调启动TLR4诱导的全身 发炎。怀孕期间使用阿片类药物会导致肠道生态失调，这与多样性和 婴儿微生物组的分类学。综上所述，产前阿片类药物暴露可能会导致 新生儿肠道微生物失调，导致全身炎症。令人惊讶的是，益生菌疗法确实 不能完全减弱阿片类药物诱导的免疫功能障碍，这表明其他机制在发挥作用 全身炎症反应综合征后的免疫抑制表型，如巨噬细胞的表观遗传修饰。H3K9me2 SIRS后巨噬细胞IL-1β和肿瘤坏死因子α启动子的修饰已被普遍描述； 长期使用阿片类药物的成人模型在很大程度上将他们对这些修饰的分析局限于脑组织， 有希望的是，H3K9me2修饰在几个大脑区域的差异表达已经被显示出来。vt.给出 这些初步发现，本项目的目的是在肠道细胞水平上研究这些修饰 巨噬细胞。我假设，产前接触阿片类药物会导致微生物失调，这 有助于全身炎症；此外，阿片类药物诱导的持续全身炎症 出生后改变肠道巨噬细胞的表观基因组，从而进一步维持免疫 功能障碍。到目前为止，还没有关于产前阿片类药物暴露如何影响肠道的研究报道。 微生物组或阿片类药物诱导的炎症如何导致巨噬细胞的表观遗传修饰。因此， 该项目的目标是表征产前接触阿片类药物后的免疫功能障碍，并确定 通过以下两个具体目标建立基本机制。目标1.确定肠道的变化如何 通过检查肠道微生物，产前阿片类药物暴露后的微生物群有助于全身炎症 代谢组分，肠道屏障的完整性，以及TLR4诱导的全身炎症。目标2. 产前阿片类药物暴露后免疫功能障碍的特征及阿片类药物诱导的影响 肠道巨噬细胞对H3K9me2修饰的炎症反应。总的来说，这项建议将有助于 深入了解产前阿片类药物暴露后免疫功能障碍，并将提供第一个 对免疫功能障碍的表观遗传学和微生物组调控的洞察。

Project Summary The heightened incidence of opioid use during pregnancy in the wake of the ongoing opioid crisis motivates this targeted investigation into the impact of prenatal opioid exposure. Here, we examine the molecular mechanisms through which prenatal opioid exposure produces systemic inflammation and immunosuppressive phenotypes later in life. This result is consistent with systemic inflammatory response syndrome (SIRS) followed by the corresponding compensatory anti-inflammatory response syndrome (CARS) that restricts harmful systemic immune responses. Recent studies have implicated the role of microbial dysbiosis in systemic inflammation and epigenetic modifications of macrophages in CARS-mediated immunosuppression. Specifically, adult models have shown that following chronic opioid exposure, gut microbial dysbiosis initiates TLR4-induced systemic inflammation. Opioid use during pregnancy results in gut dysbiosis that is strongly correlated with diversity and taxonomy of the infant microbiome. Taken together, it is plausible that prenatal opioid exposure results in neonatal gut microbial dysbiosis which contributes to systemic inflammation. Surprisingly, probiotic therapy does not fully attenuate opioid-induced immune dysfunction, suggesting other mechanisms at play that contribute to immunosuppressive phenotypes following SIRS such as epigenetic modifications of macrophages. H3K9me2 modifications in the IL-1β and TNFα promoters of macrophages following SIRS have commonly been described; adult models of chronic opioid use have largely limited their analysis of these modifications to brain tissue where, promisingly, differential expression of H3K9me2 modifications in several brain regions has been shown. Given these preliminary findings, this project aims to study these modifications on a cellular level in intestinal macrophages. I hypothesize that prenatal opioid exposure results in microbial dysbiosis, which contributes to systemic inflammation; additionally, opioid-induced systemic inflammation that persists postnatally alters the epigenome of intestinal macrophages which further perpetuates immune dysfunction. To date, there have been no reported studies on how prenatal opioid exposure affects the gut microbiome or how opioid-induced inflammation leads to epigenetic modifications of macrophages. Thus, the goal of this project is to characterize immune dysfunction following prenatal opioid exposure and to define underlying mechanisms through the following two Specific Aims. Aim 1. Determine how alterations in the gut microbiome following prenatal opioid exposure contribute to systemic inflammation by examining gut microbial and metabolome composition, gut barrier integrity, and TLR4-induced systemic inflammation. Aim 2. Characterize immune dysfunction following prenatal opioid exposure and the effects of opioid-induced inflammation on H3K9me2 modifications in intestinal macrophages. Collectively, this proposal will contribute an in-depth understanding of immune dysfunction following prenatal opioid exposure and will provide the first insights into epigenetic and microbiome regulation of immune dysfunction.