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

长期饮酒对单核细胞及其祖细胞功能和表观遗传景观的影响

• 批准号: 10526150
• 负责人: Ilhem Messaoudi
• 金额: $ 13.86万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-11-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10526150
• 关键词: Impact; chronic; alcohol; consumption; functional

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.

项目摘要 大约约7％的饮酒人员从事慢性大量饮酒（CHD），这是 与增加感染的敏感性以及伤口愈合受损和组织修复有关 导致术后结果不佳。有证据表明，其中许多缺陷是由 源自髓样细胞的过度炎症反应，特别是循环单核细胞和组织 居民巨噬细胞。这些数据主要是由来自健康的单核细胞的体外研究产生的 供体或细胞系被治疗高剂量的乙醇。但是，CHD影响的基础机制 体外研究无法完全理解，因为免疫细胞在多细胞中发挥其功能 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背景下异常炎症反应的机制的理解 仍然不完整。为了解决这些知识差距，我们建议利用恒河猴的猕猴模型 自愿乙醇自我给药，可以准确反映人类生理学并概括人类 饮酒行为。使用此模型，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布线，导致对LP的异常反应 刺激。但是，此重编程和表观遗传机制的功能含义 控制它仍然未知。重要的是，因为单核细胞是恒定的短寿命循环细胞 这些观察结果来自骨髓的重生，表明造血生态位的扰动。 对造血祖细胞的初步单细胞分析表明，分化潜力转向 与酒精相比，更多成熟的髓样祖细胞。但是，祖细胞中这种观察结果与 他们在血液和组织中的分化状态尚不清楚。在此应用程序中，我们建议测试 慢性饮酒的假设重编程了单核细胞的表观遗传景观 骨髓中的祖细胞会产生循环的单核细胞 炎症反应。我们将首先检查冠心对循环的功能重编程的影响 单核细胞，实施测定以测试其迁移，吞噬细胞的能力并生成适当的代谢 回答。接下来，我们将研究刺激对单核细胞表观遗传景观的影响 评估染色质的可及性和特定组蛋白修饰的丰度。最后，我们将确定 CHD对骨骼分化潜力，转录组激活和表观遗传学的影响 骨髓髓样祖细胞并将这些数据与从外周血单核细胞获得的数据相结合。 该提案的完成将提供有关冠心病对骨髓和机制的影响的新颖洞察力 通过它损害免疫力和宿主防御以及设计干预措施以减轻这些不利 事件并改善免疫学结果。