Effects Of EIF3e Deficiency On Mouse Liver And Screening Of EIF3e Inhibitors

Background:eIF3,assembled by 13 subunits(eIF3a-m),is the most complex translation initiation factor.It is involved in many key steps of eukaryotic translation initiation.As an important member of eIF3 family,eIF3e has been reported to be upregulated or down-regulated in different cancers.Non-alcoholic fatty liver diseases(NAFLD)include fatty accumulation,steatohepatitis,liver fibrosis,liver cirrhosis and liver cancer.The reported RNA sequencing data showed that compared with normal tissue,mRNA level of eIF3e is highly expressed in malignant liver cancer.Preliminary IHC staining results also showed that eIF3e is over expressed in liver cancer.All of these suggest that eIF3e plays an important role in liver.Methods:constructing a mouse model of specific knockout of eIF3e in the liver to explore the role of eIF3e in the liver;constructing Cellular Thermal Shift Assay and a high-throughput screening model to explore the possibility of eIF3e as a drug target.Results:In the liver-specific knockout mouse model,only mice heterozygous for eIF3e(eIF3eflox/+ Cre+)were obtained;Compared with NC group,the ALT and AST level of HE male mice tended to increase,the TP level was significantly reduced and ALB level also decreased to a certain extent.The AST level of HE female mice was higher than NC group and TP level was also lower.After 12 months,compared with NC group,58.3%of HE male mice tended to have higher weight,more abdominal fat.Liver tissue tended to be enlarged and there was accumulation of lipid droplets,but it was not developed to liver fibrosis;Compared with NC group,HE male mice showed more apoptotic cells and had a higher apoptotic ratio,also had lower energy consumption and lower fat utilization;Liver mitochondria became smaller and the matrix became deeper,suggesting that there was mitochondrial defects in the liver.In CETSA assay,we screened 90 compounds of eIF3e but found no lead compounds.In the exploration of the HST model,we calculated the Z’ to measure the deviation and variation of HTS,but the Z’ was not allowed for further screening.These two methods were instructive for the screening of small molecule inhibitors in the future.Conclusion:The results show that deficiency of eIF3e in the liver can lead to liver damage,fatty liver and hepatocyte mitochondrial defects in mice,which may be related to the occurrence and development of NAFLD.All of those provide exploratory data for functions of eIF3e in mouse tissues.