Characterization the disruption of mitochondrial function and induction of oxidative stress by SARS-CoV2

SARS-CoV2 对线粒体功能的破坏和氧化应激诱导的表征

基本信息

批准号：
10510963
负责人：
Yidong Bai
金额：
$ 19.38万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-08 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10510963
关键词：
2019-nCoV A549 ACE2 ATP Synthesis Pathway Affect Aging Antioxidants Binding Biogenesis Biology COVID-19 COVID-19 impact COVID-19 pathogenesis COVID-19 patient COVID-19 test COVID-19 treatment Cell Death Cell Line Cell model Cells Complex Defect Disease Elderly Electron Transport Energy Metabolism Engineering Epithelial Cells Etiology Exhibits Fatigue Gene Expression Genes Genome Human Immune response Investigation Knowledge Laboratories Lactate Dehydrogenase Lead Leber&apos s Hereditary Optic Neuropathy Lentivirus Light Malignant Neoplasms Measures Mediating Medicine Metabolic Metabolic Diseases Metabolic syndrome Minority Groups Mitochondria Mitochondrial DNA Mitochondrial Diseases Mitochondrial Proteins Modeling Molecular Target Mutation NADH dehydrogenase (ubiquinone)Neurodegenerative Disorders Nuclear Nuclear Import Organ Oxidative Phosphorylation Oxidative Stress Oxidative Stress Induction Oxygen Pathogenicity Patients Peptides Phosphorylation Population Process Protein Biosynthesis Proteins Proteomics Protons Public Health Reactive Oxygen Species Reporting Research Resistance Respiration Disorders Respiratory Chain Rest SARS-CoV-2 genome SARS-CoV-2 infection SARS-CoV-2 spike protein Series Serum Shortness of Breath Symptoms Syndrome System Testing Time Translations Viral Proteins Virus Virus Replication Yeasts aging population alveolar epithelium base common symptom effective therapy experimental study high risk inducible gene expression innovation insight mitochondrial DNA mutation mitochondrial dysfunction mutant novel novel strategies oligomycin sensitivity-conferring protein oxidative damage premature protein complex respiratory respiratory protein stable cell line systematic review

项目摘要

1 Populations at higher risk of severe disease from COVID-19 are the elderly and those with metabolic 2 syndromes, the populations known for compromised mitochondrial function. In addition, the most common 3 symptoms in hospitalized COVID patients are shortness of breath and fatigue, indicating deficient oxygen and 4 energy metabolism, also suggesting defective mitochondria. COVID-19 patients also have significantly 5 elevated serum lactate dehydrogenase and increases oxidative stress, pointing to a possibility of reduced 6 mitochondrial oxidative phosphorylation (OXPHOS). Together, these information leads us to consider whether 7 mitochondrial dysfunction might contribute to the pathogenesis of COVID-19. A comprehensive proteomics 8 investigation and other studies identified at least 6 mitochondrially-localized SARS-CoV-2 viral proteins which 9 were shown to interact with host cell mitochondrial proteins involved in critical OXPHOS pathways converging 10 on respiratory Complex I biogenesis. Our lab has established expertise on the investigation of mitochondrial 11 biology and mitochondrial medicine, especially on Complex I-related OXPHOS biogenesis. Over the years we 12 have developed a series of unique cell models with different types of complex I defects, including sets of cells 13 with different contents of functional complex I subunits, a set cells with different complex I assembly capacity, 14 and a set of cells carrying pathogenic mutations in complex I subunit genes, as well as an engineered system 15 to rescue complex I-related function with the introduction of a yeast Complex I counterpart NDI1 gene. These 16 models exhibit different levels of complex I subunit expression, different capacities of complex I and overall 17 respiratory machinery assembly, and different complex I and overall mitochondrial OXPHOS activities. 18 Accordingly, these cell models also exhibit different sensitivities to oxidative stress and cell death. We have 19 also initiated a line of study on the effect of viruses on mitochondria and consequent implications on human 20 diseases. In addition, we have achieved to obtain 1.Inducible expression which could turn on and off the 21 SARS-CoV-2 proteins in our cell models at proper levels; 2.Multiple genes expression which can express 22 multiple SARS-CoV-2 proteins targeting one or multiple OXPHOS pathways simultaneously in our cell models; 23 3.Establishment of A549-hACE2 cell, where a human alveolar epithelial cell line, A549 was transduced with 24 lentiviruses expressing human ACE2. A549-hACE2 cells readily support SARS-CoV2 infection and replication; 25 4.Generated mutant SARS-CoV2 lines which could serve as controls. These provide a unique opportunity for 26 us to utilize our unique systems and expertise to fulfill with two independent and integrated aims to study the 27 interactions between SARS-CoV2 and mitochondria, and their implications on oxidative stress and cell death, 28 both in cell models with regulated mitochondrial function and in human alveolar epithelial cell line infected with 29 SARS-CoV2. We expect these research will help identify molecular targets of SARS-CoV2 proteins in host 30 cells and will also provide novel approaches for protecting against the harmful effects of COVID-19. 31 32

1个来自Covid-19的严重疾病风险较高的人群是老年人和代谢的人群 2个综合征，是因线粒体功能而闻名的人群。另外，最常见的 3例住院的共vid患者的症状是呼吸急促和疲劳，表明氧气不足和 4能量代谢，还表明线粒体有缺陷。 Covid-19患者也有明显的 5升高的血清乳酸脱氢酶并增加氧化应激，表明可能降低 6线粒体氧化磷酸化（OXPHOS）。这些信息一起导致我们考虑是否 7线粒体功能障碍可能有助于Covid-19的发病机理。全面的蛋白质组学 8研究和其他研究发现至少6个线粒体占地的SARS-COV-2病毒蛋白，这些蛋白显示9个与临界Oxphos途径的宿主细胞线粒体蛋白相互作用 10关于呼吸复合物I生物发生。我们的实验室已经建立了有关线粒体调查的专业知识 11生物学和线粒体医学，尤其是在复杂的I相关OXPHOS生物发生上。多年来，我们 12开发了一系列具有不同类型的复杂I缺陷的独特细胞模型，包括一组单元格 13具有不同含量的功能复合物I亚基，一个具有不同复杂I组装能力的套件， 14和一组在复杂I亚基基因中带有致病突变的细胞以及工程系统 15通过引入酵母复合物I对应物NDI1基因来挽救与I相关的功能。这些 16个模型表现出不同级别的复杂I亚基表达，复杂I的不同能力和总体 17呼吸机械组件以及不同的复合物I和总体线粒体Oxphos活性。 18因此，这些细胞模型还表现出对氧化应激和细胞死亡的不同敏感性。我们有 19还启动了关于病毒对线粒体影响的研究线，并对人类产生影响 20种疾病。此外，我们已经实现了1.可诱导的表达，可以打开和关闭我们的细胞模型中的21个SARS-COV-2蛋白在适当的水平上； 2.可以表达的多个基因表达 22在我们的细胞模型中同时靶向一个或多个OXPHOS途径的多个SARS-COV-2蛋白； 23 3.建立A549-HACE2细胞，其中人肺泡上皮细胞系A549被转导 24慢病毒表达人ACE2。 A549-HACE2细胞很容易支持SARS-COV2感染和复制； 25 4.可以用作对照的生成突变体SARS-COV2线。这些提供了独特的机会 26我们利用我们独特的系统和专业知识来实现两个独立和综合的目标，以研究 27 SARS-COV2与线粒体之间的相互作用及其对氧化应激和细胞死亡的影响， 28在具有调节线粒体功能的细胞模型和人牙室上皮细胞系感染的细胞模型中 29 SARS-COV2。我们预计这些研究将有助于确定宿主中SARS-COV2蛋白的分子靶标 30个细胞，还将提供新的方法来防止Covid-19的有害作用。 31 32