Reprogramming of human umbilical cord-derived mesenchymal stromal cells into hepatocyte-like cells by over-expression of hepatocyte nuclear factor 1-alpha

POSTER PRESENTATIONS complex I inhibitor), which produced an increase in p62 only in the mitochondrial fraction. p62 levels were mediated through the transcription factor CHOP, while other well-known regulators (NFkB and Nrf2) are not involved. Western blot and RT-PCR analysis revealed that EFV enhanced expression of the NLRP3 inflammasome. Silencing of the p62 gene enhanced EFV-induced mRNA expression of the NLRP3 inflammasome and rendered hepatocytes more susceptible to EFV-induced mitochondrial damage, made evident by enhanced mitochondrial superoxide production; however, p62 silencing had no effect on LC3 expression, a key marker of autophagy. Conclusions: CHOP-induced p62 plays an autophagy-independent and protective (alleviating mitochondrial oxidative stress and exerting an anti-inflammatory action) role in an in vitro model of drug-induced hepatotoxicity characterized by mitochondrial dysfunction and ER-stress. These findings are of potential clinical relevance as they may help to understand the off-target hepatic effects of pharmacological agents. FRI-441 The S-ademetionine role in the prevention of liver injury in patients with acute leukemias in the polychemotherapy dynamics I. Skrypnyk1, G. Maslova1. 1Internal Medicine, Ukrainian Medical Stomatological Academy, Poltava, Ukraine E-mail: [email protected] Background and Aims: The polychemotherapy (PCT) is associated with a high risk of hepatotoxic reactions, that cause a review of the treatment tactics of patients with acute leukemia (AL), reduction the dose of cytotoxic drugs, increasing the interval between cycles, which reduces the effectiveness of a specific treatment. The aim – to assess the effectiveness of S-ademetionine (S-AME) in the prevention of medication liver injury in patients with AL in PCT dynamics. Methods: The study involved 82 patients with AL (61–acute myeloid leukemia (AML), 21–acute lymphoblastic leukemia (ALL)), mean age 17–72 years, men 48 (58.5%), 34 women (41.5%). The study included patients with newly diagnosed AL, ECOG I-II. The activity of alanine aminotransferase (ALT), aspartate (AST) aminotransferase, lactate dehydrogenase (LDH), alkaline phosphatase (ALP), gamma-glutamyltranspeptidase (GGT) was determined before chemotherapy and after two induction of remission cycles according to the protocols of ALL and AML treatment. Depending on the assigned treatment, the patients were divided into 2 groups: I (n = 42) – PCT + S-ademetionine (S-AME) 800 mg/day; II (n = 40) – control – PCT. Results: Before chemotherapy in AL pts of group I the ALT activity was 65.11 ± 15.08 U/L, AST – 53.1 ± 12.03 U/L, alkaline phosphatase – 232.2 ± 22.21 U/L, GGT – 65.2 ± 19.26 U/L, LDH – 815.4 ± 91.07 U/L, in the group II ALT activity was 61.9 ± 16.18 U/L, AST– 59.3 ± 14.13 U/L, alkaline phosphatase – 262.1 ± 23.01 U/L, GGT– 71.04 ± 21.76 U/L, LDH– 804.5 ± 81.27 U/L. After 2 courses of chemotherapy in pts of group I, who received the S-AME, ALT activity decreased in 2.2 times (29.9 ± 13.3 U/L), AST – in 2.3 times (22.7 ± 12.3 U/L), ALP – in 1.2 times (187.1 ± 17.6 U/L), GGT – in 1.3 times (49.5 ± 11.7 U/L), LDH – in 1.7 times (421.9 ± 51.1 U/L) ( p < 0.05), reaching the value of the norm. The hepatotoxicity was observed in 5 (11.9%) pts, which was associated with low efficiency of chemotherapy. In group II pts didn’t get S-AME and the development of hepatotoxicity was observed in 17 (42.5%) pts. In general, the ALT activity in this group was 58.08 ± 16.14 U/L, AST – 41.2 ± 15.15 U/L, ALP – 244.9 ± 23.15 U/L, GGT – 53.5 ± 17.1 U/L, LDH – 641.3 ± 51.82 U/L. Hepatotoxic reactions in the comparison groups were characterized by the development of cytolytic and cholestatic syndromes of moderate activity. Conclusions: Thus, in order to prevent the development of hepatotoxic reactions in the polychemotherapy dynamics in pts with AL S-AMe 800 mg/day can be recommended.

FRI-442 Reprogramming of human umbilical cord-derived mesenchymal stromal cells into hepatocyte-like cells by over-expression of hepatocyte nuclear factor 1-alpha K. Buyl1, R.M. Rodrigues1, V. Rogiers1, J. De Kock1, T. Vanhaecke1 and In Vitro Toxicology & Dermato-Cosmetology. 1Department of In Vitro Toxicology & Dermato-Cosmetology, Faculty of Medicine and Pharmacy, Vrije Universiteit Brussel, Brussels, Belgium E-mail: [email protected] Background and Aims: Stem cell technology holds great promise for the generation of human liver-based in vitro models to screen out hepatotoxic drug candidates. Recent research showed that human umbilical cord-derived mesenchymal stromal cells (hUC-MSCs) could represent a valuable cell population to generate human hepatocyte-like cells as they already express several key liver-specific transcription factors as well as hepatic progenitor markers. However, they still lack the hepatocyte nuclear factors 1-alpha (HNF1a) and 4alpha (HNF4a), indispensable for their reprogramming towards hepatocyte-like cells. Therefore, in this study, we aimed to investigate the influence of HNF1a lentiviral over-expression on the phenotype of hUC-MSCs. Methods: hUC-MSCs were transduced with HNF1a-encoding lentiviral vector using a multiplicity of infection of 1. Further, HNF1atransduced hUC-MSCs were cultured in hepatogenic differentiation medium during 21 days. Undifferentiated and differentiated hUCMSCs were used as control conditions. Whole genome microarray analysis was performed using Affymetrix microarray technology. Analysis of the transcriptomics results was carried out by using the Partek Genomics Suite and the Affymetrix Transcriptome Analysis Console. Gene expression was compared to that of human primary hepatocytes (HuHep). Statistical analysis was performed using twoway ANOVA followed by a bonferroni post hoc test (p < 0.05). Results: We found that the expression of the nuclear receptor retinoid X receptor (RXR) gamma and the nuclear transcription factor HNF4a, in HNF1a-transduced hUC-MSCs, were significantly upregulated compared to the control conditions. This expression was even higher than found in HuHep. The same was observed for the uridine 5′-diphospho-glucuronosyltransferase (UGT) 1A family, important phase II biotransformation enzymes. Further, a significant upregulation was observed compared to the control conditions for the serum proteins alpha-foetoprotein (AFP), alpha1-antitrypsin (A1-AT), the phase I biotransformation enzymes cytochrome P450 (CYP) 1A2 and CYP2A6 and the drug transporter multidrug resistance protein (MDR) 1. Conclusions: We believe that this strategy might therefore be suitable to generate hepatocyte-like cells necessary for the development of functional human liver-based in vitro models for pharmacotoxicological purposes. However, additional experiments, including liver functionality assays are required to further characterize these hUC-MSCHNF1a-derived hepatocyte-like cells. FRI-443 Free cholesterol accumulation in liver sinusoidal endothelial cells exacerbates acetaminophen hepatotoxicity in mice, T. Teratani1, K. Tomita2, T. Suzuki1, H. Furuhashi2, R. Irie3, S. Hida4, Y. Okada2, C. Kurihara2, H. Ebinuma1, N. Nakamoto1, H. Saito5, T. Hibi1, S. Miura2, R. Hokari2, T. Kanai1. 1Department of Internal Medicine, Keio University School of Medicine, Tokyo; 2Department of Internal Medicine, National Defense Medical College, Saitama; 3Department of Pathology, National Center for Child Health and Development, Tokyo; 4Department of Molecular Oncology, Institute for Biomedical Sciences, Shinshu University Graduate School of Medicine, Matsumoto; 5Graduate School of Pharmeceutical Sciences, Keio University Faculty of Pharmacy, Tokyo, Japan E-mail: [email protected] Background and Aims: Acute acetaminophen (APAP)-induced liver injury is the most frequent cause of acute liver failure in the USA. As

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Journal of Hepatology 2017 vol. 66 | S333–S542