Effect of PPARy Ligands on Alcohol-Induced Alveolar Macrophage Oxidative Stress

PPARy 配体对酒精诱导的肺泡巨噬细胞氧化应激的影响

基本信息

批准号：
8728705
负责人：
Samantha M. Yeligar
金额：
$ 11.37万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2015-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8728705
关键词：
2,4-thiazolidinedione Address Adult Respiratory Distress Syndrome Alcohol abuse Alcohols Alveolar Cell Alveolar Macrophages Animal Model Apoptotic Applications Grants Area Attenuated Award Biology Biostatistical Methods Cell Line Chronic Clinical Research Clinical Trials Data Disease Endothelial Cells Enzymes Ethanol Experimental Models Faculty Functional disorder Future Goals Grant In Vitro Infectious Agent Inflammation Mediators Ingestion Institution Intranasal Administration Investigation Klebsiella pneumonia bacterium Knockout Mice Laboratories Ligands Liver Lung Lung Capacity Lung diseases Manuscripts Mediating Mentors Mentorship Methods MicroRNAs Microbe Molecular Molecular Biology Techniques Mus NADPH Oxidase NADPH Oxidase 1 Nuclear Hormone Receptors Oxidative Stress PPAR gamma Patients Peroxisome Proliferator-Activated Receptors Phagocyte Bactericidal Dysfunction Phagocytosis Phase Pioglitazone Play Positioning Attribute Preparation Production Proteins Publications Reactive Oxygen Species Recording of previous events Regulation Research Research Personnel Respiratory Burst Respiratory Tract Infections Risk Role Solid Source Specificity Staging Techniques Therapeutic Therapeutic Intervention Thiazolidinediones Training Transgenic Mice Translations Universities Up-Regulation Writing alcohol exposure alcohol research career career development chronic alcohol ingestion clinically relevant diabetic patient experience human disease human subject immune function improved in vivo in vivo Model insulin sensitivity interest killings mRNA Transcript Degradation macrophage meetings mouse model novel novel therapeutic intervention post-doctoral training pre-doctoral problem drinker programs public health relevance receptor responsible research conduct rosiglitazone skills transcription factor

项目摘要

DESCRIPTION (provided by applicant): Chronic alcohol abuse increases patients' risk of developing Acute Respiratory Distress Syndrome (ARDS) and respiratory infections. In alveolar macrophages (AMs), NADPH oxidase (Nox) 1, Nox2, and Nox4 are critical sources of reactive oxygen species (ROS), and Nox2 is essential for the respiratory burst involved in killing microbes after phagocytosis. However, excessive ROS production suppresses phagocytosis. Chronic alcohol ingestion increases Nox enzyme levels, leading to AM oxidative stress and dysfunction. These alcohol-induced derangements can be reversed by treatment with peroxisome proliferator-activated receptor gamma (PPAR?) ligands, such as pioglitazone and rosiglitazone. In these studies, the PI will elucidate the molecular mechanisms that modulate alcohol-induced AM Nox expression and activity by studying microRNAs (miRs): Nox1-related miR-1264, Nox2-related miR-107, and Nox4-related miRs-363 and -92a/b (Aim 1). Then, the PI will examine how PPAR? ligands attenuate these miRs to reverse alcohol-mediated AM Nox1, Nox2, and Nox4 expression, oxidative stress and compromised phagocytosis (Aim 2). These hypotheses will be investigated by using a mouse model of chronic alcohol consumption, an in vitro ethanol exposed mouse AM cell line, MH-S, and AMs isolated from human subjects. The objective of the studies outlined in this application is to demonstrate that targeting PPAR? constitutes a novel therapeutic approach to ameliorate alcohol-induced AM dysfunction. If successful, these investigations could have considerable translational impact on the management of patients with a history of alcohol abuse by setting the stage for future clinical studies. The PI's focus in alcohol research began during her pre-doctoral dissertation project investigating the detrimental effects of chronic alcohol abuse in the liver, focusing on mechanisms underlying the participation of ROS and inflammatory mediators that alter liver endothelial cell and macrophage function. During post- doctoral training in the laboratories of Drs. Brown and Hart, she acquired additional expertise with numerous molecular biology techniques and with animal models of chronic alcohol ingestion. During the K99 mentored phase of the proposed project, the applicant will further expand her repertoire of skills by receiving hands-on training in: a) techniques to perform and characterize Klebsiella pneumonia bacterial challenges in the airways of mouse models to assess alveolar macrophage phagocytosis in vivo, b) methods to directly deliver PPAR? ligand therapeutics to the lungs of mouse models using intranasal administration, and c) skills required to develop and manage colonies of knockout and transgenic mice. These skills will be acquired through the execution of the proposed studies with the assistance and training of the mentors' labs during the first two years of the proposed project period. The focus of these studies will permit a natural extension of the candidate's interest in the regulation of miRs in the context of chronic alcohol ingestion. t is anticipated that her additional expertise in this area will naturally promote her growing independence from her mentors into the R00 independent phase. During the proposed award, in addition to didactic courses in current molecular biology techniques and biostatistical methods, the PI will gain non-laboratory skills important for her career development as an independent research investigator by participating in seminars and activities related to the responsible conduct of research, laboratory management, and faculty career development. Support from this K99/R00 grant will provide the PI an outstanding opportunity to expand and consolidate her experimental and laboratory skills and support her career goal to become an independent investigator and obtain a faculty position at an academic institution. The likelihood that she will achieve these goals is supported by planned mentorship from well-established investigators, the abundant scientific opportunities within the Emory University Alcohol and Lung Biology Center, and a hypothesis-driven application exploring novel mechanisms of an important and clinically relevant pathophysiological disorder. The proposed program will permit the PI to build her publication record, collect critical preliminary data for subsequent grant applications, present research findings at national meetings, and gain experience in manuscript writing and grant preparation. Thus, this K99/R00 application provides an excellent opportunity to advance the career of a talented and promising investigator.

描述（由申请人提供）：慢性酒精滥用会增加患者患急性呼吸窘迫综合征（ARDS）和呼吸道感染的风险。在肺泡巨噬细胞（AMS）中，NADPH氧化酶（NOX）1，NOX2和NOX4是活性氧的关键来源（ROS），而NOX2对于杀死吞噬细胞后杀死微生物的呼吸道爆发至关重要。但是，过量的ROS产生抑制了吞噬作用。慢性酒精摄入增加了NOX酶水平，导致AM氧化应激和功能障碍。这些酒精诱导的紊乱可以通过过氧化物酶体增殖物激活的受体伽马（PPAR？）配体（例如吡格列酮和罗斯加列酮）的处理来逆转。在这些研究中，PI将通过研究microRNA（miRS）：与NOX1相关的miR-1264，与NOX2相关的miR-107和NOX4相关的MIRS-363和-92A/B（AIM 1）来调节酒精诱导的NOX表达和活性的分子机制。然后，PI会检查PPAR的情况吗？配体会衰减这些miR，以反向酒精介导的AM NOX1，NOX2和NOX4表达，氧化应激和受损的吞噬作用（AIM 2）。这些假设将通过使用长期饮酒的小鼠模型，体外乙醇暴露的小鼠AM细胞系，MH-S和从人类受试者分离的AMS进行研究。本应用程序中概述的研究的目的是证明针对PPAR？构成一种新型的治疗方法，可改善酒精诱导的AM功能障碍。如果成功，这些调查可能会通过为未来的临床研究奠定阶段，从而对患有酗酒史的患者的管理产生重大的翻译影响。 PI在酒精研究中的重点始于其论文前论文项目，研究了慢性酒精滥用在肝脏中的有害影响，重点是ROS和炎症介体的参与机制，这些机制改变了改变肝内皮细胞和巨噬细胞功能的炎症介体。在博士实验室的博士后培训期间。布朗和哈特（Brown and Hart），通过大量分子生物学技术以及慢性酒精摄入的动物模型获得了更多的专业知识。在拟议项目的K99指导阶段中，申请人将通过接受动手培训来进一步扩大她的技能曲目：使用鼻内给药和c）开发和管理基因敲除和转基因小鼠菌落所需的技能对小鼠模型的肺的配体治疗。这些技能将通过在拟议项目期间的头两年内在指导者实验室的协助和培训中执行拟议的研究来获得这些技能。这些研究的重点将允许在慢性酒精摄入的情况下自然扩展候选人对MIR的兴趣。预计她在这一领域的额外专业知识自然会促进她从导师到R00独立阶段的独立性。在拟议的奖励期间，除了当前分子生物学技术和生物统计学方法的教学课程外，PI还将通过参与与负责任的研究，实验室管理和教师职业发展相关的活动和活动来获得对她作为独立研究研究者的职业发展的非实验室技能。这项K99/R00赠款的支持将为PI提供一个出色的机会，可以扩展和巩固她的实验和实验室技能，并支持她的职业目标，以成为独立研究员并在学术机构获得教师职位。她实现这些目标的可能性得到了良好的研究人员的计划指导，埃默里大学酒精和肺生物学中心的大量科学机会以及假设驱动的应用程序探索了重要和临床相关的病理生理障碍的新机制。拟议的计划将允许PI建立她的出版记录，收集关键的初步数据以进行后续赠款申请，在国家会议上进行研究结果，并获得手稿写作和赠款准备的经验。因此，该K99/R00应用程序提供了一个极好的机会，可以推进才华横溢且有前途的调查员的职业。