Alcohol-induced Skeletal Muscle Insulin Insensitivity in SIV/HIV: Myotube-derived Extracellular Vesicle-mediated Mechanisms

摘要 此Ruth L.Kirschstein NRSA F30应用程序的主要目的是为 为申请者的学术医疗生涯做好准备。申请者的大部分职业发展将来自 从高度优先的艾滋病毒/艾滋病共病相关研究中脱颖而出，她将获得宝贵的机会 使用已建立的慢性酗酒(CBA)临床前模型进行高质量研究 猴免疫缺陷病毒(SIV)感染。艾滋病毒携带者使用高风险酒精的可能性是前者的两倍 (PLWH)，就像在一般人口中一样。接受抗逆转录病毒治疗(ART)的PLWH患者的生活接近正常 预期，但代谢性并存的患病率增加，需要进行研究 在HIV背景下研究代谢失调的病理生理学机制。来自我们小组的研究 已表明高危饮酒和CBA与PLWH和PLWH的胰岛素/血糖动力学受损有关 分别在感染SIV的猕猴中。骨骼肌(SKM)是胰岛素依赖型葡萄糖的主要来源 摄取，临床前研究表明，酒精降低SKM的胰岛素敏感性。初步数据 为这一应用而产生的结果表明，CBA在SKM中扰乱了胰岛素信号通路中的蛋白质， 进一步表明酒精导致SKM功能障碍。我们小组之前的工作表明 CBA介导的肌管改变反映了在SKM中看到的，我们将利用培养的肌管来反映 在我们提议的实验中，SKM发生了变化。细胞外小泡(EVS)介导细胞变化 通过运输生物活性货物，包括microRNAs(MiRNAs)， 细胞。MiRNAs是一种小的非编码RNA，调节基因表达，并通过 CBA与胰岛素抵抗。初步和先前的数据表明，CBA导致了 血浆EVS和SIV感染猕猴SKM特异性miRNA-206与miRNA-206有关 在SKM中，胰岛素依赖的葡萄糖摄取。综上所述，我们的初步数据和出版的文献支持 假设酒精介导的肌管细胞外小泡miRNA货运量的改变 降低SIV感染患者的肌管胰岛素敏感性。拟议的研究将采用各种不同的 使用两个具体目标来检验这一假说的最先进技术：(1)CBA-管理改变肌管- 在SIV感染的猕猴中获得EV图谱，以及(2)CBA注射后肌管来源的EV miRNA货物 感染SIV的猕猴会降低肌管胰岛素敏感性。拟议研究的结果将提供 深入了解EVS在酒精介导的SIV感染中SKM胰岛素不敏感中的作用。此外，建议的 该项目旨在说明EVS在改善酒精介导的SKM功能障碍方面的治疗潜力。使用 一个强大的指导团队致力于培养一名全面发展的内科科学家，完成拟议的 高优先级艾滋病毒/艾滋病共病相关研究的培训计划将确保申请者做好准备 开始从事学术医学的职业生涯。

Abstract The primary purpose of this Ruth L. Kirschstein NRSA F30 application is to provide the groundwork that will prepare the applicant for an academic medical career. Much of the applicant’s career development will come from work in High Priority HIV/AIDS comorbidity-related research, and she will have the invaluable opportunity to carry out high quality research using a well-established preclinical model of chronic binge alcohol (CBA) and simian immunodeficiency virus (SIV) infection. At-risk alcohol use is twice as likely in people living with HIV (PLWH) as it is in the general population. PLWH on antiretroviral therapy (ART) have a near normal life expectancy, but there is an increase in prevalence of metabolic comorbidities, requiring a need for research studying pathophysiological mechanisms of metabolic dysregulation in the context of HIV. Studies from our group have shown that at-risk alcohol use and CBA is associated with impaired insulin/glucose dynamics in PLWH and in SIV-infected macaques, respectively. Skeletal muscle (SKM) is a major site of insulin-dependent glucose uptake, and preclinical studies indicate that alcohol decreases SKM insulin sensitivity. Preliminary data generated for this application suggest that CBA dysregulates proteins in the insulin signaling pathway in SKM, further indicating that alcohol leads to SKM dysfunction. With previous work from our group demonstrating that CBA-mediated alterations in myotubes reflect what is seen in SKM, we will utilize cultured myotubes to reflect changes seen in SKM in our proposed experiments. Extracellular vesicles (EVs) mediate cellular changes in pathologic and therapeutic conditions by transporting bioactive cargo, including microRNAs (miRNAs), between cells. miRNAs are small non-coding RNAs that regulate gene expression and are differentially expressed by CBA and insulin resistance. Preliminary and previous data suggests that CBA leads to lower expression of the SKM specific miRNA-206 in plasma EVs and the SKM of SIV-infected macaques, and miRNA-206 is implicated in SKM insulin-dependent glucose uptake. Taken together, our preliminary data and published literature support the hypothesis, that alcohol-mediated alterations in myotube extracellular vesicle miRNA cargo contribute to decreased myotube insulin sensitivity in SIV infection. The proposed study will employ a wide variety of state-of- the-art techniques to test the hypothesis using two specific aims: (1) CBA-administration alters myotube- derived EV profile in SIV-infected macaques, and (2) myotube-derived EV miRNA cargo of CBA-administered SIV-infected macaques decreases myotube insulin sensitivity. Findings from the proposed studies will provide insight on the role of EVs in alcohol-mediated SKM insulin insensitivity in SIV infection. Additionally, the proposed project aims to illustrate the therapeutic potential of EVs in improving alcohol-mediated SKM dysfunction. With a strong mentoring team committed to developing a well-rounded physician scientist, completion of the proposed training plan in High Priority HIV/AIDS comorbidity-related research will ensure that the applicant is ready to embark on a career in academic medicine.