Disclosures: Fabio Marra – Advisory Committees or Review Panels:

Disclosures: Fabio Marra – Advisory Committees or Review Panels: Abbott; Consulting: Bayer Healthcare; Grant/Research Support: ViiV Massimo Pinzani – Advisory Committees or Review Panels: Intercept Pharmaceutical, Silence Therapeutic, Abbot; Consulting: UCB; Speaking and Teaching: Gilead, BMS The following

people have nothing to disclose: Jose Macias-Barragan, Jose Vera-Cruz, Jesus Garcia-Banuelos, David A. Lopez-de la Mora, Cibeles San-chez-Roque, Krista Rombouts, Juan Armendáriz-Borunda Background: Nonalcoholic Fatty Liver Disease (NAFLD) is a heritable and prevalent disease, affecting about 30% of the population. A characteristic feature of NAFLD is hepatic steatosis, the presence of excess fat (mostly triglycerides (TG)) in the liver. Using genome wide association analysis (GWAS) we identified genetic variants in PNPLA3 and Wnt pathway GCKR, and near LYPLAL1 that associate with population based hepatic steatosis. How these variants result in increased liver steatosis is not known. Here we aim to characterize the genetic mechanism by which genetic variants at

these loci may affect nearby genes to result in hepatic triglyceride accumulation. Methods: HuH-7 and HepG2 liver cell lines were infected with lentiviruses expressing wildtype PNPLA3, learn more GCKR, and LYPLAL1 as well as the mutants PPP1R3B(I148M) and GCKR(P446L) or with shRNAs to PNPLA3, GCKR, and LYPLAL1 and stably expressing cell lines were selected. Overexpression/knockdown was quantified using Western/Northern blotting analysis. Stable cell lines were loaded with oleic acid, hepatic steatosis was measured using LipidTOXTM however (Life Technology), and total cellular triglyceride was quantified using a Triglyceride Determination Kit (Sigma-Aldrich). Results: Overexpression of wildtype and to a larger

extent mutant PNPLA3 but not knockdown of PNPLA3 resulted in increased steatosis/TG accumulation. Over-expression of wildtype GCKR but not mutant GCKR resulted in increased steatosis/TG accumulation whereas knockdown of GCKR also resulted in increased steatosis/TG accumulation. Overexpression of LYPLAL1 resulted in decreased steatosis/TG accumulation and knockdown in increased steatosis/TG accumulation. Conclusions: These results suggest that variants in PNPLA3 exert their effect through an increase/gain-of-function mechanisms, those in GCKR and near LYPLAL1 likely through a loss-of-function mechanism. Disclosures: The following people have nothing to disclose: Yue Chen, Andrew W. Tai, Elizabeth K. Speliotes Background and aims: We developed an optimized diet-induced mouse model that produces robust inflammation and fibrosis. Using this model we investigated the changes of proin-flammatory transcripts expressed in liver and fat tissues.

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