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.
This study is highly relevant for public health because it will identify how certain natural chemicals present in the human diet control the production of Nuclear factor-erythroid 2 related factor 2 (Nrf2). Nrf2 is considered the master regulator of the body owns antioxidant response and promoting the activation of Nrf2 has been shown to be beneficial for the treatment of non- alcoholic fatty liver disease. Thus, this study has tremendous potential to accelerate the development of novel activators of Nrf2 for this disease.
| Zhang, Hanghang; Pandey, Somnath; Travers, Meghan et al. (2018) Targeting CDK9 Reactivates Epigenetically Silenced Genes in Cancer. Cell 175:1244-1258.e26 |
| Perez-Leal, Oscar; Barrero, Carlos Alberto; Merali, Salim (2017) Pharmacological stimulation of nuclear factor (erythroid-derived 2)-like 2 translation activates antioxidant responses. J Biol Chem 292:14108-14121 |
