Screening Small Molecular To Normalize Mutation Related Keratin Structure Disruption And Provide Protection From Liver Injury

Background There are a lot of different etiologies could result in the human liver injury, and among those the Hepatitis B is the major reason in China. Regardless of Hepatitis B or other etiologies, like DILI, NASH or biliary diseases, there could be huge individual differences of the outcomes from the patients with the same etiology, which suggests that other potential reasons may contribute to it. Keratin is the biggest family of the intermediate filaments, and is found play an important role in the liver injury. We hypothesis that keratin maybe one of the reason to trigger individual differences of the liver injury, and it also will provide a new treatment area of the liver injury through keratin. Liver specific keratin 8 and 18 mutation could result in the keratin structure disruption in the hepatic cells, and make the cells easy to be apoptotic and the transgenic mice predispose to the liver injury. Exploring the treatment for the keratin 8/18 mutation-related predisposition to liver injury may could provide a new way for the treatment for the liver injury.Aim In this study, we will focus on the liver specific keratin 8/18 mutation mediated structure disruption, using the drug screening to find small molecular that could stable the mutated keratin structure, thereby rescue the keratin function and provide the protect for the cells and mice. This study will provide a new treatment idea to the Intermediate filament-related diseases and more importantly, it will give the different outcomes of the liver injury a potential reason and show a new treatment area for the liver injury.Methods We will use the imaged-based drug screening, which relied on the structure changes seen by immunoflorensence after the drug use. The hits will be confirmed in the mutated cells and transgenic mice, and we will also detect the mechanism of the small molecular function.Results In this paper, we demonstrate that PKC412, which is the multi-kinase inhibitor, could stable the mutated K18 R90 C structure and provide the protection in the mutated cells and transgenic mice form apoptosis and acute liver injury. And we also show that the mechanism of PKC412 is to induce the hypophosphorylation of non-muscle myosin IIA and trigger its binding to the keratins.Conclusions We defined PKC412 as the compound that could alleviate mutated keratin structure, and provide protection for the mutated cells and mice from apoptosis and acute liver injury. The mechanism of under that is because that PKC412 could increase keratin and NMHC-IIA binding, which also related to NMHC-IIA’s phosphorylation.