Turning on Nrf2 translation for chronic liver disease

开启 Nrf2 翻译治疗慢性肝病

基本信息

批准号：
9326288
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
项目类别：
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/9326288
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 Luc Gene Luciferases Luteolin Messenger RNA 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 molecular targeted therapies non-alcoholic fatty liver novel nuclear factor-erythroid 2 prevent public health relevance response transcription factor

项目摘要

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（由适用提供）：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（NRF2）的转化的机制 - 一种重要的分子靶标，用于治疗或预防肝脏的慢性疾病。氧化应激是参与非酒精性脂肪肝疾病（NAFLD）的生成和永久化的过程，影响高达30％NRF2的最普遍的肝病是该细胞自身防御系统针对氧化条件的主要调节剂，并且研究的广泛体系表明，NRF2的活性可能会导致NRF2的活性促进生理策略。我们最近发现了一种新的机制，该机制在人类细胞的生理条件下衡量了NRF2的翻译。该机制尚未完全理解，取决于位于开放式阅读框架中的mRNA序列的一部分，如果序列被同义词密码子取代突变，则可以减轻其抑制作用。更重要的是，序列的这一部分能够防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记者，以验证以下假设：一些已知的自然抗氧化剂增加了NRF2活性，可以通过促进其翻译来起作用。我们确定了类黄酮替代蛋白是NRF2翻译的潜在诱导剂，并通过多种技术证实了这一点。我们还发现了其他类黄酮（例如槲皮素和叶黄素）的类似特性。现在的挑战是确定允许这些类黄酮激活NRF2的翻译的蛋白质靶标，也看看是否是否人类饮食中存在的其他类黄酮具有这种特性。为了执行此操作，我们建议基于最前沿蛋白质组学技术进行RNA-蛋白相互作用研究，并使用荧光素酶报告基因筛选500个类黄酮的库。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

CRISPR/Cas9-Based Screening of FDA-Approved Drugs for NRF2 Activation: A Novel Approach to Discover Therapeutics for Non-Alcoholic Fatty Liver Disease.

DOI：
10.3390/antiox12071363
发表时间：
2023-06-29
期刊：
ANTIOXIDANTS
影响因子：
7
作者：
Li, James;Arest, Sandra;Olszowy, Bartlomiej;Gordon, John;Barrero, Carlos A.;Perez-Leal, Oscar
通讯作者：
Perez-Leal, Oscar

Identification of Inhibitors of Tubulin Polymerization Using a CRISPR-Edited Cell Line with Endogenous Fluorescent Tagging of β-Tubulin and Histone H1.