An Integration of Transcriptomics And Proteomics Reveals That miR‐33‐5p Is A Key Player of Lipogenesis Signaling in Fructose‐Induced NAFLD in C57BL/6N Mice

Abstract

Fructose is a strong risk factor of non‐alcoholic fatty liver disease (NAFLD), but limited studies have investigated mechanisms by which microRNAs (miRNA) are interlinked in the etiology of NAFLD. Here, we aimed to investigate the significance of miRNAs in fructose‐induced NAFLD conditions through integration of liver transcriptomics and proteomics. C57BL/6N mice (male and female) were fed with either water (n=14) or 34% fructose (n=14) for 6 wk ad libitum. After, liver transcriptomics and proteomics datasets were integrated to narrow down key signaling pathways and to find upstream regulator(s) that are responsible for fructose induced NAFLD in which srebp1 was found to be the most significant upstream regulator. In an attempt to explore regulatory roles of miRNAs in NAFLD, Ingenuity Pathway Analysis software miRNA database was used to list miRNAs that are known to interact with the upstream regulator, srebp1. Out of seven final miRNA candidates, miR‐33a was selected after our qPCR validation in mice liver and AML12 mouse hepatocytes that were treated with fructose. In order to further validate the interaction between miR‐33a and srebp1, AML12 hepatocytes were transfected with the miR‐33a; in this, srebp1, and its downstream proteins, acc1 and scd1 were all decreased by miR‐33a transfection whereas suppression of miR‐33a, achieved by anti‐miR‐33a transfection, restored those protein expression changes. Altogether, based on our unbiased approaches as well as in vitro validation experiments, fructose induces NALFD conditions via suppressing miR‐33a in liver thereby increasing srebp1 and its downstream lipogenic proteins, acc1 and scd1.Support or Funding InformationThis work was supported by the University of Arkansas, VPRED Start‐up fund (JKK, and JHP). Support has been provided in part by the Arkansas Biosciences Institute, a partnership of scientists from Arkansas Children's Hospital, Arkansas State University, the University of Arkansas‐Division of Agriculture, the University of Arkansas, Fayetteville, and the University of Arkansas for Medical Sciences. The Arkansas Biosciences Institute is the major research component of the Arkansas Tobacco Settlement Proceeds Act of 2000 (JKK, JHP, KEB, and BCK). This work was also supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. NRF‐2015R1C1A1A01053746; JHL) as well as the China Scholarship Council (JT).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.