Alcohol and calorie-dense diet-mediated hepatic mitochondrial dysregulation

1篇摘要 2这位露丝·L. kirschstein 3职业生涯申请人的职业发展将在很大程度上来自于酒精，高浓度酒精， 4脂肪，高蔗糖暴露。申请人将有宝贵的机会进行高质量的 5项研究使用了长期酗酒（CBA）给药的成熟临床前猕猴模型， 6肝细胞的体外HepaRG球状体细胞培养模型。酒精相关的肝损伤是导致 7终末器官疾病，以及酒精和高热量饮食对肝脏病理生理学的综合影响， 八是要做好特色。一些关于酒精或高脂肪饮食相关肝损伤的研究表明， 9线粒体氧化磷酸化（OXPHOS）和重塑（生物发生和动力学）是 10减少或中断响应酒精或高脂肪。来自实验室的数据显示CBA介导的 11降低参与线粒体生物发生、OXPHOS和融合的基因的肝脏表达， 12过氧化物酶体增殖物激活受体-g共激活因子-1b（PGC-1b）和线粒体融合蛋白-2（MFN2） 13只猕猴此外，从短期高脂肪肝的肝活检中产生的初步数据， 14只高蔗糖（HFS）饮食（HFSD）喂养的猕猴显示脂质相关基因表达增加， 15积累实验室以前的工作表明，酒精会使线粒体功能失调， 16骨骼肌和免疫细胞的重塑和生物能量学。总体而言，初步数据和 17篇已发表的文献提供了酒精介导的线粒体适应导致终末器官损伤的证据。 18因此，拟议项目的假设是酒精和HFSD一起破坏线粒体重塑， 19降低OXPHOS，增加肝脏中的脂质积累，并促进PGC-1b表达 20改善肝细胞中的这些变化。使用来自CBA或给予HFSD的溶媒的猕猴的肝脏 21在多个时间点和酒精和HFS处理的肝细胞的体外模型，申请人旨在测试 酒精和HFS破坏肝线粒体重塑，减少肝OXPHOS，并增加肝细胞的OXPHOS。 23肝脂质蓄积。此外，无论是否促进肝细胞PGC-1b表达， 24这些酒精和HFs介导的改变将被确定。拟议研究的结果将提供 25深入了解HFSD背景下酒精引起的代谢失调。此外，该项目旨在 26显示PGC-1b过表达增加了线粒体生物发生、融合和OXPHOS，并具有 27改善酒精和HFSD引起的肝损伤的肝脏健康的潜力。拥有强大的指导团队 28致力于发展一个全面的医生科学家，完成拟议的培训计划，在酒精 29与滥用有关的研究将确保申请人有能力在学术医学领域取得成功。 NRSA F30申请是准备申请人的学术医疗

1 Abstract 2 The goal of this Ruth L. Kirschstein 3 career. The applicant’s career development will come in large part from work in hepatic models of alcohol, high 4 fat, and high sucrose exposure. The applicant will have the invaluable opportunity to conduct high quality 5 research using a well-established preclinical macaque model of chronic binge alcohol (CBA) administration and 6 an in vitro HepaRG spheroid cell culture model of hepatocytes. Alcohol-related liver injury is a leading cause of 7 end-organ disease, and the combined impact of alcohol and a calorie-dense diet on liver pathophysiology has 8 yet to be well characterized. Several studies of either alcohol- or high fat diet-related liver injuries demonstrate 9 that mitochondrial oxidative phosphorylation (OXPHOS) and remodeling (biogenesis and dynamics) are 10 decreased or disrupted in response to alcohol or high fat. Data from the laboratory showed CBA-mediated 11 decreased hepatic expression of genes involved in mitochondrial biogenesis, OXPHOS, and fusion such as 12 peroxisome proliferator-activated receptor-g coactivator-1b (PGC-1b) and mitofusin-2 (MFN2) in control diet-fed 13 macaques. Additionally, preliminary data generated for this proposal from liver biopsies of short-term high fat, 14 high sucrose (HFS) diet (HFSD) fed macaques showed increased expression of genes involved in lipid 15 accumulation. Previous work from the laboratory demonstrated that alcohol dysregulates mitochondrial 16 remodeling and bioenergetics in skeletal muscle and immune cells. Taken together, preliminary data and 17 published literature provide evidence of alcohol-mediated mitochondrial adaptations leading to end-organ injury. 18 Thus, the hypothesis of the proposed project is that alcohol and HFSD together disrupt mitochondrial remodeling, 19 decrease OXPHOS, and increase lipid accumulation in the liver, and that promoting PGC-1b expression 20 ameliorates these changes in hepatocytes. Using livers from CBA or vehicle administered HFSD-fed macaques 21 at multiple timepoints and an in vitro model of alcohol and HFS-treated hepatocytes, the applicant aims to test 22 that alcohol and HFS disrupt hepatic mitochondrial remodeling, decrease hepatic OXPHOS, and increase 23 hepatic lipid accumulation. Further, whether or not promoting hepatocyte PGC-1b expression will ameliorate 24 these alcohol and HFS-mediated alterations will be determined. Findings from the proposed studies will provide 25 insight into metabolic dysregulation caused by alcohol in the context of HFSD. Additionally, the project aims to 26 show that PGC-1b overexpression increases mitochondrial biogenesis, fusion, and OXPHOS and has the 27 potential to improve liver health in alcohol- and HFSD-induced liver injury. With a strong mentoring team 28 dedicated to developing a well-rounded physician scientist, completion of the proposed training plan in alcohol 29 abuse-related research will ensure that the applicant is equipped for a successful career in academic medicine. NRSA F30 application is to prepare the applicant for an academic medical