Impact of chronic alcohol consumption on the functional and epigenetic landscapes of monocytes and their progenitors

项目摘要 大约7%的饮酒者患有慢性重度饮酒（CHD）， 与感染易感性增加以及伤口愈合和组织修复受损有关 导致术后效果不佳。有证据表明，这些缺陷中有许多是由 源自骨髓细胞，特别是循环单核细胞和组织的过度炎症反应， 常驻巨噬细胞这些数据主要来自体外研究，其中来自健康人的单核细胞 供体或细胞系用高剂量乙醇处理。然而，冠心病影响的潜在机制 不能通过体外研究完全理解，因为免疫细胞在多细胞环境中执行其功能。 酒精具有广泛影响的环境。由于缺乏利用可靠的体内模型的研究， 我们对CHD中异常炎症反应机制的理解 仍然不完整。为了解决这些知识差距，我们建议利用恒河猴模型， 自愿乙醇自我管理，准确反映人体生理学和概括复杂的人类 饮酒行为使用这个模型，我们的实验室最近证明了CHD导致转录和 循环单核细胞和脾巨噬细胞的表观遗传重新连接，导致对LPS的异常反应 刺激.然而，这种重编程和表观遗传机制的功能影响， 控制它仍然未知。重要的是，由于单核细胞是在恒定的免疫应答下短寿命的循环细胞， 这些观察结果表明造血生态位的扰动。 造血祖细胞的初步单细胞分析表明，分化潜能向 更成熟的骨髓祖细胞然而，在祖细胞中的观察结果与 它们在血液和组织中的分化状态仍不清楚。在本申请中，我们建议测试 慢性酒精消耗重编程单核细胞表观遗传景观假说 骨髓中的祖细胞产生循环单核细胞， 炎症反应。我们将首先研究冠心病对循环系统功能重编程的影响， 单核细胞，实施测定以测试其迁移、吞噬和产生适当代谢产物的能力， 应答接下来，我们将通过以下方式研究刺激对单核细胞表观遗传景观的影响： 评估染色质可及性和特异性组蛋白修饰的丰度。最后，我们将确定 CHD对骨的分化潜能、转录组激活和表观遗传重连的影响 并将这些数据与从外周血单核细胞获得的数据整合。 这项提案的完成将为CHD对骨髓生成的影响及其机制提供新的见解 它损害了免疫力和宿主防御以及设计干预，以减轻这些不利影响， 事件和改善免疫结果。

PROJECT SUMMARY Approximately ~7% of alcohol-consuming individuals engage in chronic heavy drinking (CHD), which is associated with increased susceptibility to infections as well as impaired wound healing and tissue repair resulting in poor post-operative outcomes. Evidence suggests that many of these defects are mediated by excessive inflammatory responses originating from myeloid cells, notably circulating monocytes and tissue- resident macrophages. These data were primarily generated from in vitro studies where monocytes from healthy donors or cell lines are treated high doses of ethanol. However, the mechanisms underlying the effects of CHD cannot be fully understood by in vitro studies because immune cells carry out their functions in a multicellular environment in which alcohol has widespread effects. Due to a lack of studies utilizing reliable in vivo models, our understanding of the mechanisms underlying aberrant inflammatory responses in the context of CHD remains incomplete. To address these knowledge gaps, we propose to leverage a rhesus macaque model of voluntary ethanol self-administration that accurately mirrors human physiology and recapitulates complex human drinking behavior. Using this model, our lab has recently demonstrated that CHD results in transcriptional and epigenetic rewiring of circulating monocytes and splenic macrophages, resulting in aberrant responses to LPS stimulation. However, the functional implications of this reprogramming and the epigenetic mechanisms controlling it remain unknown. Importantly, because monocytes are short-lived circulating cells under constant repopulation from the bone marrow, these observations suggest perturbations of the hematopoietic niche. Preliminary single-cell analyses of hematopoietic progenitors point to a shift in differentiation potential towards more mature myeloid progenitors with alcohol. However, a link between this observation in progenitor cells and their differentiated states in blood and tissue remains unclear. In this application, we propose to test the hypothesis that chronic alcohol consumption reprograms the epigenetic landscape of monocyte progenitors in the bone marrow giving rise to circulating monocytes poised towards a hyper- inflammatory response. We will first examine the impact of CHD on functional reprogramming of circulating monocytes, implementing assays to test their ability to migrate, phagocytose, and generate proper metabolic responses. We will next examine the effect of stimulation on the monocyte epigenetic landscape through assessment of chromatin accessibility and abundance of specific histone modifications. Finally, we will determine the effect of CHD on the differentiation potential, transcriptome activation, and epigenetic rewiring of bone marrow myeloid progenitors and integrate these data with those obtained from peripheral blood monocytes. Completion of this proposal will provide novel insight into the impact of CHD on myelopoiesis and mechanisms by which it compromised immunity and host defense as well as design interventions to mitigate these adverse events and improve immunological outcomes.