Alcohol-induced myomiR dysregulation: mechanisms of impaired skeletal muscle regeneration in SIV/HIV

项目总结/摘要 该NIH指导研究科学家发展奖（K01）应用程序的目的是提供 在三个关键领域开展高度优先研究所需的额外培训和研究技能 HIV/AIDS相关的合并症。具体来说，候选人将熟练地进行基本和 转化研究的重点是酒精介导的代谢并发症， 接受抗逆转录病毒治疗的艾滋病毒感染者。申请人是一个有前途的 她是一位在干细胞和信号传导研究方面有着良好记录的科学家，她现在建议将其整合到 酒精引起的表观遗传改变综合全面的发展计划 研究技能将建立候选人在表观遗传分析，生物统计学和翻译方面的专业知识 research.这项研究计划将集中在阐明酒精引起的损害的机制 SIV/HIV中骨骼肌（SKM）再生的研究。成肌细胞分离自慢性酗酒（CBA）- 施用的猕猴显示出降低的肌原性基因表达和分化成 在体外培养时，在缺乏胰岛素的情况下，观察到肌源性分化受损。 体外酒精暴露，这表明这些成肌细胞有一种“记忆”， 变化初步证据还表明，在骨骼肌中，肌肉特异性microRNA（myomiRs）失调， CBA给药SIV感染猕猴的SKM导致CBA损害SKM的假设 SIV/HIV感染通过肌原基因转录的表观遗传修饰再生。这 假设将通过三个具体目标进行严格检验。第一个目标将检验CBA的预测， 导致成肌细胞myomiR表达和分化潜能降低。为了解决这个问题， 候选人将利用她在干细胞培养和分子技术方面的现有技能。此外，她还将在 在原位杂交技术方面接受过培训，以确定成肌细胞培养物中myomiR的表达。的 第二个目标将测试CBA诱导的成肌细胞组蛋白修饰有助于 肌原性分化潜能降低。使用分子技术，将myomiRs的靶基因 经验证，将确定组蛋白脱乙酰酶（HDAC）活性和组蛋白修饰。第三个目标 将在一组人的循环中建立改变的myomiR表达的临床相关性 艾滋病毒/艾滋病感染者（PL W H A）和酒精使用障碍（AUD）。myomiR的循环水平将被 与AUD和肌肉力量（握力计）相关，以确定其作为 肌源性功能受损的指标。拟议和未来研究的总体目标是确定 目标是通过物理（运动）或药物干预改善骨骼肌功能。的 综合翻译和跨学科培训计划将支持候选人的进展， 在酒精诱导的骨骼肌表观遗传调节领域的独立工作 再生潜力，并将有助于研究培训所需的劳动力进行高 在非人类灵长类动物和艾滋病毒感染者中开展艾滋病毒/艾滋病相关研究的优先事项。

Project Summary/Abstract The objective of this NIH Mentored Research Scientist Development Award (K01) application is to provide additional training and research skill sets in three critical areas needed to conduct high priority research in HIV/AIDs-associated comorbidities. Specifically, the candidate will be skilled to conduct basic and translational research focused on the alcohol-mediated metabolic complications prevalent in the prematurely aged individuals infected with HIV on antiretroviral therapy. The applicant is a promising scientist with a track record of research in stem cells and signaling, which she now proposes to integrate with the field of alcohol-induced epigenetic alterations. The integrated and comprehensive plan to develop research skills will build the candidate’s expertise in epigenetic analysis, biostatistics, and translational research. The research proposal will focus on elucidating the mechanisms of alcohol-induced impairment of skeletal muscle (SKM) regeneration in SIV/HIV. Myoblasts isolated from chronic binge alcohol (CBA)- administered macaques show decreased myogenic gene expression and potential to differentiate into myotubes when cultured in vitro. Impaired myogenic differentiation was observed in the absence of in vitro alcohol exposure, suggesting that these myoblasts had a “memory” that preserved gene expression changes. Preliminary evidence also suggests dysregulated muscle-specific microRNAs (myomiRs) in the SKM of CBA-administered SIV-infected macaques leading to the hypothesis that CBA impairs SKM regeneration in SIV/HIV infection through epigenetic modifications of myogenic gene transcription. This hypothesis will be critically tested by three specific aims. The first aim will test the prediction that CBA results in decreased myoblast myomiR expression and differentiation potential. To address this, the candidate will use her existing skills in stem cell culture and molecular techniques. In addition, she will be trained in in situ hybridization techniques for determining expression of myomiRs in myoblast cultures. The second aim will test the prediction that CBA-induced myoblast histone modifications contributes to decreased myogenic differentiation potential. Using molecular techniques target genes of myomiRs will be validated, histone deacetylase (HDAC) activity, and histone modifications will be determined. The third aim will establish the clinical relevance of altered myomiR expression in the circulation in a cohort of persons living with HIV/AIDS ( P L W H A ) with alcohol use disorders (AUD). Circulating levels of myomiRs will be correlated with AUD and muscle strength (hand grip dynamometer) to establish their potential use as indicators of impaired myogenic function. The overarching goal of the proposed and future studies is to identify targets to improve skeletal muscle function using physical (exercise) or pharmacological interventions. The integrated translational and transdisciplinary training plan will support the candidate’s progression to an independent productive career in the field of alcohol- induced epigenetic modulation of skeletal muscle regenerative potential and will contribute to the research training of the workforce required to conduct high priority HIV/AIDS related research in non-human primates and PLWHA.