Bio distribution and function of alcohol-induced exRNA

描述（由申请人提供）：细胞外miRNAs（ex-miRs）在细胞间和器官间通讯中具有独特的作用。我们的初步数据表明，人类的急性酒精狂饮和小鼠的慢性酒精给药均导致细胞外micoRNA（ex-miR）水平增加。我们发现，炎症的主要调节因子miR-155在小鼠肝脏中被慢性酒精增加，这与肝脏炎症和循环中miR-155的增加相关。我们的研究结果还表明，人类和小鼠的急性酒精狂欢导致“装载”miR-155的循环外泌体数量显著增加。基于这些发现，我们假设急性酒精狂欢和慢性酒精消耗都导致外泌体释放到循环中，并且外泌体的频率和ex-miR和/或ex-miR“货物”在急性和慢性酒精消耗后可能不同。使用miR-155缺陷小鼠，我们发现加载有成熟miR-155的外泌体将可检测水平的miR-155递送至受体小鼠的血浆和肝脏，表明外泌体包装的ex-miR的广泛生物分布。我们假设外泌体为转移酒精诱导的功能有效的ex-miR提供了独特的载体。我们推测外泌体中酒精诱导的ex-miR对肝脏和/或其他器官具有功能性作用。具体目标是：1。为了使用miR-155缺陷小鼠测试急性酒精狂欢和慢性酒精施用对血浆和不同器官中外泌体包装的miR-155的体内生物分布的影响，2.为了描述外泌体包装的miR-155在体外和体内的生物学效应，3.评估急性酒精狂欢和慢性酒精消耗对小鼠和人类循环ex-miR谱的影响。这些实验将表征小鼠中急性酒精狂饮后与慢性酒精使用相比的血浆ex-miR谱，并且这将与急性酒精狂饮后来自人类志愿者的数据进行比较。我们的实验将揭示外泌体递送的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.