Duplex miR-223 and Exosomes in Sepsis

败血症中的双链 miR-223 和外泌体

基本信息

批准号：
9195740
负责人：
Guo-Chang Fan
金额：
$ 30万
依托单位：
UNIVERSITY OF CINCINNATI
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-01-01 至 2018-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9195740
关键词：
Affect Animals Apoptosis Attenuated Blood Blood Vessels Bone Marrow Cardiac Cardiac Death Cardiac Myocytes Cardiomyopathies Cardiovascular system Cause of Death Cells Cessation of life Clinical Clinical Research Code Critical Illness Data Development Engineering Functional disorder Goals Heart Heart failure Human Impairment Inflammation Inflammation Mediators Inflammatory Inflammatory Response Injection of therapeutic agent Injury Interleukin-1 Interleukin-6 Knock-out Knockout Mice Knowledge Lead Ligands Ligation Macrophage Inflammatory Protein-1 Mediating Mediator of activation protein Membrane Proteins Mesenchymal Stem Cells MicroRNAs Mission Modeling Morbidity - disease rate Mus Muscle Cells Myocardial Myocardial dysfunction Myocardium Nitric Oxide Synthase Pathway interactions Patients Peritoneal Peritoneal Fluid Play Production Public Health Puncture procedure RNA Reagent Reducing Agents Research Role STAT3 gene Semaphorin-3A Sepsis Septic Shock Serum Signal Pathway Source Stress Study of serum TNF gene TNFRSF1A gene Testing Therapeutic Therapeutic Agents Therapeutic Effect Therapeutic Intervention Tissues Transgenic Mice Treatment Efficacy United States National Institutes of Health Untranslated Regions Up-Regulation Work base chemokine cytokine disability exosome improved insight interest macrophage mortality mouse model nanovesicle novel novel therapeutics overexpression public health relevance receptor receptor expression screening septic survival outcome therapeutic evaluation tool transcription factor translational study

项目摘要

DESCRIPTION (provided by applicant): Cardiovascular dysfunction is a major contributor to sepsis-induced death in critically ill patients. Despite decades of intensive study, the basic mechanisms remain elusive. In particular, therapeutic interventions aimed at a single mediator or pathways of inflammation have failed to improve cardiac function and survival outcome in sepsis. Thus, continuing search for effective therapeutic reagents which target multiple factors is greatly needed. Recent identified microRNAs may provide new targets for effective sepsis therapy. Our latest work has discovered, for example, that: 1) miR-223 (-5p & -3p) are significantly reduced in mouse hearts after severe sepsis; 2) global loss of duplex miR-223 aggravated sepsis-induced cardiac dysfunction and mortality; and 3) miR-223-5p inhibited the expression of TNFR1 and Sema3A (a ligand of TLRs), while miR-223-3p repressed the expression of IL-6, STAT3, MIP-1�MIP-2 in macrophages and myocytes. Given that each of these molecules can potentially contribute to sepsis-induced cardiac dysfunction and mortality, it will be significant to test the effects of global elevation of duplex miR-223 on sepsis-caused injury. Our pilot data also indicate that: 1) lesser amounts of duplex miR-223 are encased within exosomes isolated from the blood of septic mice (referred to as septic exosomes), compared to healthy exosomes and 2) septic exosomes impaired cardiomyocyte contractility and stimulated macrophages to release pro-inflammatory cytokines (i.e. TNF-�IL-6). Thus, it will be important to test whether duplex miR-223 is a critical limiting factor whose reduction contributes to septic exosome-induced detrimental effects on cardiac function and mortality. Finally, an important translational question is raised: are there any positive therapeutic effects of engineering exosomes to contain extra miR-223 on sepsis? In this proposal, we hypothesize that global elevation of duplex miR-223 limits sepsis-induced cardiac dysfunction and mortality through targeting diverse mediators at multiple levels, and loss of duplex miR-223 contributes to septic exosome-triggered cardiac dysfunction. These ideas will be tested by pursuing three specific aims: 1) Using a transgenic mouse model, we will test whether global elevation of duplex miR-223 attenuates sepsis-induced myocardial depression and mortality; 2) Utilizing duplex miR-223-null exosomes and septic exosomes isolated from the blood of miR-223-knockout mice and CLP-operated mice to test whether septic exosome-mediated cardiac dysfunction and mortality is ascribed to the reduction of duplex miR-223; and 3) We will use bone marrow-derived mesenchymal stem cells (MSCs) as a source of miR-223-exosomes to determine the therapeutic effects of duplex-miR-223-engineered exosomes on sepsis-induced cardiac dysfunction and mortality. Together, the proposed studies are expected to unveil a previously unrecognized role of duplex miR-223 in sepsis-induced heart failure. Additionally, they are expected to provide novel insights that lead to the development of original exosome-based therapeutic strategies for reducing septic death.

描述（由适用提供）：心血管功能障碍是重症患者败血症导致死亡的主要因素。尽管进行了数十年的深入研究，但基本机制仍然难以捉摸。特别是，针对单个介体或炎症途径的治疗干预措施未能改善败血症的心脏功能和生存结果。那，继续寻找针对多个因素的有效治疗剂的是非常需要。最近确定的microRNA可能为有效的败血症提供新的靶标。例如，我们的最新工作发现：1）MiR-223（-5p＆-3p）在严重的败血症后，小鼠心脏中有显着降低； 2）双链miR-223的全球损失累计败血症诱导的心脏功能障碍和死亡率； 3）miR-223-5p抑制了TNFR1和SEMA3A（TLR的配体）的表达，而miR-223-3p抑制了巨噬细胞和心肌细胞中IL-6，STAT3，MIP-1 MIP-2的表达。鉴于这些分子中的每一个都可能导致败血症诱导的心脏功能障碍和死亡率，因此测试双链miR-223全球升高对败血症引起的损伤的影响非常重要。我们的试点数据还表明：1）与健康外泌体相比，从败血小鼠血液（称为败血症外泌体）中分离出的外泌体中的较少数量的双链miR-223被包裹在外泌体中（称为败血症外泌体），而2）败血性外泌体造成了心脏模拟性的收缩性和刺激性乳状噬菌体（释放proflampophines cytokines）（I. cytokines）。这是重要的是要测试双工miR-223是否是关键的限制因素，其还原有助于败血性外泌体引起的对心脏功能和死亡率的有害影响。最后，提出了一个重要的翻译问题：工程外泌体对败血症中含有额外的miR-223有任何积极的治疗作用吗？在这项提案中，我们假设双链miR-223的全球升高通过以多个水平靶向潜水员介体来限制了败血症诱导的心脏功能障碍和死亡率，并且双工miR-223的丧失有助于败血性外体触发的心脏功能障碍。这些想法将通过追求三个特定目的来测试：1）使用转基因小鼠模型，我们将测试全球双工miR-223的升高是否减弱了败血症引起的心肌抑郁症和死亡率； 2）利用从miR-223-敲除小鼠的血液中分离出的双链miR-223-null外泌体和化粪池外泌体和CLP锻炼小鼠的血液，以测试败血性外部外部介导的心脏功能障碍和死亡率是否分配给Duplex miR-223的减少； 3）我们将使用骨髓衍生的间充质干细胞（MSC）作为miR-223-诊断的来源，以确定双链肌223工程外泌体对脓毒症诱导的心脏功能障碍和死亡率的治疗作用。共同预计拟议的研究将揭示以前未认识到的双工miR-223在败血症诱导的心力衰竭中的作用。此外，他们有望提供新颖的见解，从而导致原始基于外泌体的治疗策略来减少败血症死亡。