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: 10440492
• 负责人: Ilhem Messaoudi
• 金额: $ 34.35万
• 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/10440492
• 关键词: ATAC-seq; Abstinence; Address; Adverse event; Alcohol consumption; Alcoholic Liver Diseases; Alcohols; Animals; Area; Bacterial Infections; Biological Assay; Biological Response Modifiers; Biology; Blood; Blood specimen; Bone Marrow; Cardiovascular Diseases; Cell Line; Cells; Chromatin; Chronic; Chronic Disease; Communicable Diseases; Complex; Data; Defect; Development; Dose; Environment; Epigenetic Process; Ethanol; Etiology; Event; Failure; Female; Gene Expression; Genetic Transcription; Goals; Health; Heavy Drinking; Hematopoietic; Hematopoietic stem cells; Heterogeneity; Homeostasis; Host Defense; Housing; Human; Immune; Immune Tolerance; Immunity; Immunocompetence; Immunologics; Immunology procedure; Impaired wound healing; In Vitro; Individual; Infection; Inflammation Mediators; Inflammatory; Inflammatory Response; Intercistronic Region; Intervention; Knowledge; Link; Macaca; Macaca mulatta; Mediating; Metabolic; Metabolism; Modeling; Myelogenous; Myeloid Cells; Myelopoiesis; Natural Immunity; Outcome; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Phenotype; Physiology; Postoperative Period; Predisposition; Promoter Regions; Reporting; Research; Risk; Sampling; Self Administration; Testing; Tissues; Viral Respiratory Tract Infection; Virus Diseases; alcohol availability; alcohol effect; antimicrobial; cancer type; chronic alcohol ingestion; cytokine; drinking behavior; epigenome; experimental study; fitness; histone modification; immune activation; improved; in vivo; in vivo Model; insight; macrophage; male; migration; monocyte; next generation sequencing; nonhuman primate; novel; pathogen; peripheral blood; progenitor; programs; response; single cell analysis; single-cell RNA sequencing; stem cells; therapy design; tissue repair; transcriptome; transcriptomics; wound healing

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.

项目总结 大约7%的饮酒者长期酗酒(CHD)，这是 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，这些缺陷中的许多是由 来自髓系细胞的过度炎症反应，特别是循环单核细胞和组织- 常驻巨噬细胞。这些数据主要是从体外研究中产生的，其中来自健康人群的单核细胞 捐赠者或细胞系接受高剂量的乙醇治疗。然而，CHD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中执行它们的功能 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， 我们对冠心病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 自愿乙醇自我给药，准确反映人体生理并概括复杂的人类 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排，导致对内毒素的异常反应 刺激。然而，这种重新编程的功能含义和表观遗传机制 控制它仍然是个未知数。重要的是，单核细胞在恒定状态下是短暂的循环细胞。 从骨髓中重新繁殖，这些观察表明造血生态位受到了干扰。 对造血祖细胞的初步单细胞分析表明，分化潜能向 更多成熟的骨髓祖细胞与酒精。然而，在祖细胞中观察到的这种联系与 它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 长期饮酒重塑单核细胞表观遗传格局的假说 骨髓中的祖细胞能产生循环中的单核细胞，并趋向于高... 炎症反应。我们将首先研究CHD对循环功能重新编程的影响 单核细胞，进行检测以测试其迁移、吞噬和产生适当代谢的能力 回应。接下来，我们将研究刺激对单核细胞表观遗传格局的影响 染色质可及性和特异组蛋白修饰丰度的评估。最后，我们将确定 冠心病对骨的分化潜能、转录组激活和表观遗传重排的影响 并将这些数据与从外周血单核细胞获得的数据进行整合。 这一提议的完成将为CHD对骨髓生成的影响和机制提供新的见解 通过它损害免疫力和宿主防御，以及设计干预措施来减轻这些不利影响 并改善免疫学结果。