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：
8990972
负责人：
Guo-Chang Fan
金额：
$ 30.71万
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/8990972
关键词：
Affect Animals Apoptosis Attenuated Basic Science 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 Health Heart Heart failure Human Inflammation Inflammatory Inflammatory Response Injection of therapeutic agent Injury Interleukin-1 Interleukin-6 Knock-out Knockout Mice Knowledge Lead Life 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 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 Untranslated Regions Up-Regulation Work base chemokine cytokine disability exosome improved insight interest macrophage meetings mortality mouse model nanovesicle novel novel therapeutics overexpression receptor receptor expression screening septic survival outcome 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（由申请人提供）：心血管功能障碍是危重患者脓毒症引起的死亡的主要原因，尽管经过数十年的深入研究，但其基本机制仍然难以捉摸，特别是针对单一介质或炎症途径的治疗干预措施尚未明确。未能改善脓毒症的心脏功能和生存结果，因此，继续寻找针对多种因素的有效治疗试剂。最近发现的 microRNA 可能为有效的脓毒症治疗提供新的靶标，例如：1) 严重脓毒症后小鼠心脏中的 miR-223 (-5p 和 -3p) 显着减少；双链体 miR-223 的整体缺失会加重脓毒症引起的心功能障碍和死亡率；3) miR-223-5p 抑制 TNFR1 和 Sema3A（TNFR1 的配体）的表达。 TLRs），而 miR-223-3p 抑制巨噬细胞和肌细胞中 IL-6、STAT3、MIP-1、MIP-2 的表达，鉴于这些分子中的每一种都可能导致脓毒症引起的心脏功能障碍和死亡。对于测试双链体 miR-223 整体升高对脓毒症引起的损伤的影响具有重要意义。我们的试验数据还表明：1）内含较少量的双链体 miR-223。从脓毒症小鼠的血液中分离出的外泌体（称为脓毒症外泌体），与健康外泌体相比，2）脓毒症外泌体会损害心肌细胞收缩力并刺激巨噬细胞释放促炎细胞因子（即TNF-αIL-6）。对于测试双链体 miR-223 是否是一个关键限制因素非常重要，其减少会导致脓毒症外泌体对心脏功能和死亡率产生不良影响。提出了一个重要的转化问题：改造外泌体以包含额外的 miR-223 对脓毒症是否有任何积极的治疗作用？在这项提案中，我们解决了双链体 miR-223 的整体升高通过针对不同的目标限制脓毒症引起的心脏功能障碍和死亡率的问题。这些想法将通过三个具体目标进行测试：1）使用转基因小鼠模型，我们将测试。双链体 miR-223 的整体升高是否会减轻脓毒症引起的心肌抑制和死亡率；2) 利用从 miR-223 敲除小鼠和 CLP 手术小鼠的血液中分离出的双链体 miR-223 缺失外泌体和脓毒症外泌体来测试是否脓毒症外泌体介导的心脏功能障碍和死亡率归因于双链体 miR-223 的减少；3）我们将使用骨；骨髓间充质干细胞（MSC）作为 miR-223 外泌体的来源，以确定双链 miR-223 工程外泌体对脓毒症引起的心脏功能障碍和死亡率的治疗效果，预计这些研究将共同揭晓。此外，它们有望提供新的见解，从而促进基于外泌体的原始治疗策略的发展。以减少脓毒症死亡。