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

项目摘要/摘要 NIH指导性研究科学家发展奖(K01)申请的目标是提供 需要在三个关键领域提供额外的培训和研究技能，以开展高度优先的研究 艾滋病毒/艾滋病相关的合并症。具体地说，应聘者将熟练地进行基本和 翻译研究的重点是酒精介导的代谢并发症在 接受抗逆转录病毒治疗的感染艾滋病毒的早老者。应聘者是个有前途的人。 在干细胞和信号方面有研究记录的科学家，她现在建议整合这些研究 在酒精引起的表观遗传改变领域。统筹谋划发展 研究技能将培养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 研究。该研究计划将侧重于阐明酒精所致损伤的机制。 SIV/HIV中骨骼肌(SKM)再生的研究。从慢性酗酒(CBA)中分离的成肌细胞- 给猕猴注射后，肌源性基因表达降低，并有可能分化为 肌管在体外培养时。在没有In的情况下观察到肌源性分化受损。 体外酒精暴露，表明这些成肌细胞具有保存基因表达的“记忆” 改变。初步证据还表明，肌肉特异的microRNAs(MyomiRs)在 CBA对SIV感染猕猴SKM的影响导致CBA损害SKM的假说 SIV/HIV感染通过肌源性基因转录的表观遗传修饰而再生。这 假设将通过三个具体目标进行批判性检验。第一个目标是检验CBA的预测 结果成肌细胞myomiR表达降低，分化潜能降低。为了解决这个问题， 应聘者将运用她在干细胞培养和分子技术方面的现有技能。此外，她还将成为 训练有素的原位杂交技术用于测定成肌细胞培养中的肌红蛋白受体的表达。这个 第二个目标将测试CBA诱导的成肌细胞组蛋白修饰有助于 肌源性分化潜能降低。使用分子技术靶向myomiRs基因将是 将确定有效的组蛋白脱乙酰酶(HDAC)活性和组蛋白修饰。第三个目标 将确定在一组人群中循环中的myomiR表达改变的临床相关性 艾滋病毒/艾滋病患者(P L，女，A)合并酒精使用障碍(AUD)。循环中的MyomiRs水平将是 与AUD和肌肉力量(手握力测力计)相关，以确定其潜在用途 肌原功能受损的指标。拟议和未来研究的首要目标是确定 通过身体(运动)或药物干预改善骨骼肌功能的目标。这个 综合翻译和跨学科培训计划将支持应聘者晋升为 在酒精诱导的骨骼肌表观遗传调控领域的独立生产生涯 再生潜力，并将有助于对需要进行高技能培训的劳动力进行研究培训 在非人类灵长类动物和PLWHA中优先开展与艾滋病毒/艾滋病有关的研究。

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.