Turning on Nrf2 translation for chronic liver disease

 描述（由申请人提供）：本研究旨在表征允许类黄酮调节核因子红细胞2相关因子2（Nrf 2）翻译的机制，Nrf 2是治疗或预防慢性肝脏疾病的重要分子靶点。氧化应激是参与非酒精性脂肪性肝病（NAFLD）的发生和持续的过程，NAFLD是影响高达30%的美国人口的最普遍的肝病。Nrf 2是细胞自身防御系统对抗氧化条件的主要调节剂，广泛的研究表明，用植物化学物质增加Nrf 2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，在人体细胞的生理条件下调节Nrf 2的翻译。该机制尚未完全理解，其依赖于位于开放阅读框中的mRNA序列的一部分，并且如果该序列被同义密码子取代而突变，则其抑制作用减轻。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，我们能够使用这种荧光素酶构建体作为报告基因来验证这一假设，即一些已知的增加Nrf 2活性的天然抗氧化剂可能通过促进其翻译而起作用。我们鉴定了类黄酮芹菜素作为Nrf 2翻译的有效诱导剂，并通过多种技术证实了这一点。我们还发现了其他类黄酮如槲皮素和木犀草素的类似性质。现在的挑战是确定允许这些黄酮类化合物激活Nrf 2翻译的蛋白质靶点， 存在于人类饮食中的其它类黄酮具有这种特性。为了执行这一点，我们建议进行RNA-蛋白质相互作用的研究，基于最先进的蛋白质组学技术，也使用荧光素酶报告筛选500类黄酮的库。

 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.