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

基本信息

批准号：
10252788
负责人：
Sloan Alexandra Lewis
金额：
$ 1.98万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2021-12-09
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10252788
关键词：
ATAC-seq Abstinence Activities of Daily Living Address Alcohol consumption Alcoholic Liver Diseases Alcohols Animals Bacterial Infections Behavior Binding Biological Assay Biology Blood Bone Marrow Cardiovascular Diseases Cell Line Cells ChIP-seq Chromatin Chronic Communicable Diseases Complex Data Defect Development Dose Epigenetic Process Ethanol Etiology Event Failure Female Gene Expression Genetic Transcription Goals Health Heavy Drinking Hematopoiesis Hematopoietic Hematopoietic stem cells Heterogeneity Human Immune Immunologics Immunology procedure Impaired wound healing Impairment In Vitro Incidence Individual Infection Inflammation Inflammation Mediators Inflammatory Inflammatory Response Innate Immune System Intercistronic Region Knowledge Laboratories Link Macaca Macaca mulatta Mediating Metabolic Metabolism Modeling Myelogenous Myeloid Cells Myeloid Progenitor Cells Outcome Peripheral Peripheral Blood Mononuclear Cell Phenotype Physiology Postoperative Period Predisposition Promoter Regions Reporting Research Risk Sampling Self Administration Testing Tissues United States Viral Respiratory Tract Infection Virus Diseases alcohol availability alcohol effect alcohol exposure antimicrobial antimicrobial peptide cancer type chronic alcohol ingestion drinking behavior experimental study granulocyte-monocyte progenitors histone modification 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 tissue repair transcription factor transcriptome transcriptomics wound healing

项目摘要

Project Summary: Alcohol consumption is widespread in the United States with ~7% of alcohol-consuming individuals engaging in heavy alcohol use. It is well established that chronic heavy drinking (CHD) 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. However, many of the current studies rely on in vitro exposure of monocytes from healthy donors or cell lines to high doses of ethanol. 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. In order 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 and the epigenetic mechanisms controlling this reprogramming 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 phenotype in progenitor cells and their differentiated states in blood 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 then examine the effect of stimulation on the monocyte epigenetic landscape through assessment of chromatin accessibility and differential binding of histone modifications with alcohol. Finally, we will determine the effect of CHD on the differentiation potential, transcriptome activation, and epigenetic rewiring of bone marrow myeloid progenitors. We will perform functional assays on monocytes derived in vitro from granulocyte/monocyte progenitors and integrate these data with those obtained from peripheral monocytes. Further, scRNA-Seq analysis and epigenetic assessment of the myeloid progenitors will allow us to determine the specific effects of alcohol on the bone marrow compartment and how this leads to hyper-inflammatory, epigenetically reprogrammed peripheral monocytes. Completion of this proposal will expand our knowledge of the immunological effects of alcohol consumption on hematopoiesis.

项目摘要：在美国，饮酒量普遍存在，约有7％的饮酒人员从事大量的酒精使用。据确定，慢性重饮酒（CHD）与增加有关对感染的敏感性以及伤口愈合受损和组织修复，导致术后不良结果。有证据表明，其中许多缺陷是由过度炎症反应介导的起源于髓样细胞，尤其是循环的单核细胞和组织居民巨噬细胞。但是，很多目前的研究依赖于从健康供体或细胞系的单核细胞体外暴露到高剂量的乙醇。由于缺乏利用可靠的体内模型的研究，我们对基本机制的理解在CHD背景下，异常的炎症反应仍然不完整。为了解决这些知识差距，我们建议利用自愿乙醇自我管理的恒河猴猕猴模型反映人类生理学并概括了复杂的人类饮酒行为。使用此模型，我们的实验室有最近证明，冠心病导致循环单核细胞的转录和表观遗传重新布线脾巨噬细胞，导致对LPS刺激的异常反应。但是，功能含义控制此重编程的控制和表观遗传机制仍然未知。重要的是，因为单核细胞是短暂的循环细胞，在骨髓的恒定重生下，这些观察结果表明造血生态位的扰动。造血的初步单细胞分析祖细胞指出，分化潜力向更成熟的髓样祖细胞转移。但是，祖细胞中这种表型与血液中的分化状态之间的联系不清楚。在此应用中，我们建议测试慢性酒精消耗重编程的假设骨髓中单核细胞祖细胞的表观遗传景观导致循环单核细胞有望朝着高炎性反应。我们将首先检查冠心病的影响循环单核细胞的功能重编程，实施测定以测试其迁移能力，吞噬细胞，并产生适当的代谢反应。然后，我们将检查刺激对通过评估组蛋白的染色质可及性和差异结合，单核细胞表观遗传景观酒精修饰。最后，我们将确定CHD对分化潜力的影响，转录组激活和骨髓髓样祖细胞的表观遗传重新布线。我们将执行功能从粒细胞/单核细胞祖细胞中衍生出的单核细胞的测定，并将这些数据与这些数据整合在一起从外周单核细胞获得。此外，髓样的SCRNA-SEQ分析和表观遗传评估祖细胞将使我们能够确定酒精对骨髓室的特定影响以及如何这导致过度炎症，表观遗传重编程的外周单核细胞。完成此操作提案将扩大我们对饮酒对造血的免疫学影响的了解。