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.

项目概要：酒精消费在美国很普遍，约 7% 的饮酒者从事酒精消费大量饮酒。众所周知，慢性大量饮酒（CHD）与增加易受感染以及伤口愈合和组织修复受损，导致术后效果不佳结果。有证据表明，许多这些缺陷是由过度炎症反应介导的起源于骨髓细胞，特别是循环单核细胞和组织驻留巨噬细胞。然而，许多目前的研究依赖于健康供体或细胞系的单核细胞在体外暴露于高剂量的乙醇。由于缺乏利用可靠的体内模型的研究，我们对潜在机制的理解冠心病背景下的异常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模型，该模型可以准确地反映了人类的生理学并概括了复杂的人类饮酒行为。我们的实验室利用这个模型最近证明，CHD 会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。然而，功能上的影响控制这种重编程的表观遗传机制仍然未知。重要的是，因为单核细胞是短命的循环细胞，不断从骨髓中重新增殖，这些细胞观察结果表明造血生态位受到干扰。造血功能的初步单细胞分析祖细胞表明，随着酒精的作用，分化潜力会转向更成熟的骨髓祖细胞。然而，祖细胞的这种表型与其在血液中的分化状态之间的联系仍然存在不清楚。在此应用中，我们建议测试以下假设：长期饮酒会重新编程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准备产生高炎症反应。我们将首先检查 CHD 对循环单核细胞的功能重编程，实施检测以测试其迁移能力，吞噬作用，并产生适当的代谢反应。然后我们将检查刺激对通过评估染色质可及性和组蛋白的差异结合来评估单核细胞表观遗传景观用酒精进行修饰。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系祖细胞的表观遗传重连。我们将执行功能对来自粒细胞/单核细胞祖细胞的体外单核细胞进行测定，并将这些数据与那些数据整合从外周单核细胞获得。此外，骨髓细胞的 scRNA-Seq 分析和表观遗传学评估祖细胞将使我们能够确定酒精对骨髓室的具体影响以及如何影响这会导致外周单核细胞过度炎症、表观遗传重编程。完成此该提案将扩大我们对饮酒对造血免疫影响的了解。