Bio distribution and function of alcohol-induced exRNA

描述(申请人提供)：细胞外miRNAs(ex-miRs)在细胞间和器官间的通讯中具有独特的作用。我们的初步数据表明，人类的急性酗酒和小鼠的长期饮酒都会导致细胞外microoRNAs(ex-miR)水平的增加。我们发现，炎症的主要调节因子miR-155在小鼠的肝脏中因慢性酒精而增加，这与肝脏炎症和循环中miR-155的增加有关。我们的结果还表明，人类和小鼠的急性酒精狂欢导致循环中携带miR-155的外体数量显著增加。基于这些发现，我们假设急性饮酒和慢性饮酒都会导致外切体释放到循环中，并且急性饮酒和慢性饮酒后外切体的频率和ex-miR和/或ex-miR“货物”可能不同。利用miR-155缺陷小鼠，我们发现携带成熟miR-155的外切体将可检测到的miR-155水平递送到受体小鼠的血浆和肝脏中，这表明外切小体包装的ex-miRs在生物上分布广泛。我们假设，外切体为转移酒精诱导的功能强大的exmiRs提供了一种独特的载体。我们推测酒精诱导的Exosome中的exmiRs对肝脏和/或其他器官有功能影响。本研究的具体目的是：1.利用miR-155基因缺陷小鼠检测急性酗酒和慢性酒精摄入对外切体包装的miR-155在血浆和不同器官中的体内分布的影响；2.研究外切体包装的miR-155在体内外的生物学效应；3.评价急性酗酒和慢性饮酒对小鼠和人循环ex-miR谱的影响。这些实验将比较急性酗酒和长期饮酒后小鼠的血浆ex-miR图谱，并将其与急性酗酒后人类志愿者的数据进行比较。我们的实验将揭示外切体传递的ex-miRNAs的生物分布、组织定位的决定因素，并揭示酒精诱导的ex-miR-155的组织靶点和功能。

DESCRIPTION (provided by applicant): Extracellular miRNAs (ex-miRs) have unique roles in cell-cell and inter-organ communications. Our preliminary data demonstrate that both acute alcohol binge drinking in humans and chronic alcohol administration in mice result in increased level of extracellular micoRNAs (ex-miR). We found that miR-155, a major regulator of inflammation, is increased by chronic alcohol in mice in the liver and this correlated with both liver inflammation and increased miR-155 in the circulation. Our results also demonstrate that acute alcohol binge in humans and in mice results in a significant increase in the number of circulating exosomes that are "loaded" with miR-155. Based on these findings we hypothesize that both acute alcohol binge and chronic alcohol consumption result in exosome release to the circulation and that that the frequency and ex-miR and/or the ex-miR "cargo" of exosomes might be different after acute and chronic alcohol consumption. Using miR-155 deficient mice we found that exosomes loaded with mature miR-155 deliver detectable levels of miR-155 to the plasma and liver of recipient mice suggesting a wide biological distribution of exosome-packaged ex-miRs. We hypothesize that exosomes provide a unique vehicle for transferring functionally potent ex-miRs induced by alcohol. We postulate that the alcohol-induced ex-miRs in exosomes have functional effects on the liver and/or in other organs. The Specific Aims are: 1. To test the effects of acute alcohol binge and chronic alcohol administration on the in vivo biodistribution of exosome-packaged miR-155 in the plasma and different organs using miR-155- deficient mice, 2. To delineate the biological effects of exosome-packaged miR-155 in vitro and in vivo, 3. To evaluate the effects of acute alcohol binge and chronic alcohol consumption on circulating ex-miR profile in mice and humans. These experiments will characterize plasma ex-miR profile after acute alcohol binge in comparison with chronic alcohol use in mice and this will be compared to data from human volunteers after acute alcohol binge drinking. Our experiments will reveal determinants of biodistribution, tissue localization of exosome-delivered ex-miRNAs, and reveal tissue targets and functions of alcohol-induced ex-miR-155.