Turning on Nrf2 translation for chronic liver disease

开启 Nrf2 翻译治疗慢性肝病

• 批准号: 8873073
• 负责人: Oscar Perez
• 金额: $ 18.41万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8873073
• 关键词: Affect; American; Antioxidants; Apigenin; Binding; Binding Proteins; Cells; Characteristics; Chemicals; Codon Nucleotides; Development; Diet; Disease; Firefly Luciferases; Flavonoids; Genetic Translation; Half-Life; Hepatocyte; Human; Laboratories; Libraries; Liver diseases; Luciferases; Luteolin; Messenger RNA; Molecular Target; Mutate; Nonesterified Fatty Acids; Open Reading Frames; Oxidative Stress; Pathogenesis; Physiological; Phytochemical; Play; Population; Prevention; Process; Production; Property; Proteins; Proteomics; Public Health; Quercetin; RNA-Protein Interaction; Regulation; Regulatory Element; Reporter; Reporter Genes; Research; Role; Signal Transduction; System; Techniques; Testing; Toxic effect; Translations; Work; base; chronic liver disease; design; dietary supplements; genetic regulatory protein; high reward; high risk; non-alcoholic fatty liver; novel; nuclear factor-erythroid 2; prevent; public health relevance; response; transcription factor

 DESCRIPTION (provided by applicant): This study is designed to characterize the mechanism that allows flavonoids to regulate the translation of Nuclear factor erythroid 2-related factor 2 (Nrf2) - an important molecular target for the treatment or prevention of chronic diseases of the liver. Oxidative stress is a process involved in the genesis and perpetuation of non-alcoholic fatty liver disease (NAFLD), the most prevalent liver disease affecting up to 30% of the American population. Nrf2 is the master regulator of the cell's owns defense system against oxidative conditions, and an extensive body of research indicates that increasing the activity of Nrf2 with phytochemicals could be a viable strategy to ameliorate the progression of NAFLD. We recently discovered a novel mechanism regulating the translation of Nrf2 under physiological conditions in human cells. The mechanism, which is not fully understood, is dependent on a portion of the mRNA sequence located in the open reading frame, and its inhibitory effect is alleviated if the sequence is mutated with synonym codon substitutions. More importantly, this portion of the sequence is able to prevent the translation of the reporter gene Firefly Luciferase. Subsequently, we were able to use this luciferase construct as a reporter to validate the hypothesis that some known natural antioxidants that increase the Nrf2 activity might work by promoting its translation. We identified the flavonoid Apigenin as a potent inducer of the translation of Nrf2 and we confirmed this via multiple techniques. We also found a similar property for other flavonoids such as Quercetin and Luteolin. The challenge now is to identify the protein targets that allow these flavonoids to activate the translation of Nrf2 and also see if other flavonoids present in the human diet share this property. To execute this, we propose performing RNA-protein interaction studies based on cutting edge proteomics techniques and also using the Luciferase reporter to screen a library of 500 flavonoids.

 描述(申请人提供)：这项研究旨在描述允许类黄酮调节核因子红系相关因子2(Nrf2)翻译的机制，Nrf2是治疗或预防慢性肝脏疾病的重要分子靶点。氧化应激是非酒精性脂肪性肝病(NAFLD)发生和持续的一个过程，NAFLD是最常见的肝病，影响多达30%的美国人口。Nrf2是细胞自身抵御氧化条件的主要调节因子，大量研究表明，用植物化学物质提高Nrf2的活性可能是改善NAFLD进展的可行策略。我们最近发现了一种新的机制，调节Nrf2在生理条件下在人类细胞中的翻译。这一机制尚不完全清楚，它依赖于位于开放阅读框架中的一部分mRNA序列，如果该序列用同义词密码子替换突变，其抑制作用就会减轻。更重要的是，这部分序列能够阻止报告基因萤火虫荧光素酶的翻译。随后，我们能够使用这个荧光素酶结构作为报告，验证了一些已知的天然抗氧化剂可能通过促进其翻译而增加Nrf2活性的假设。我们确定类黄酮类芹菜素是Nrf2翻译的有效诱导剂，并通过多种技术证实了这一点。我们还发现其他黄酮类化合物，如栎素和木犀草素也具有类似的特性。现在的挑战是确定允许这些黄酮类化合物激活Nrf2翻译的蛋白质靶标，并看看 人类饮食中存在的其他类黄酮类化合物也具有这一特性。为了实现这一点，我们建议进行基于尖端蛋白质组学技术的RNA-蛋白质相互作用研究，并使用荧光素酶报告程序来筛选500个黄酮类化合物的文库。