Noortje Ijssennagger1, Aafke W F Janssen2, Alexandra Milona1, José M Ramos Pittol1, Danielle A A Hollman1, Michal Mokry3, Bark Betzel4, Frits J Berends4, Ignace M Janssen4, Saskia W C van Mil5, Sander Kersten2. 1. Department of Molecular Cancer Research, Center for Molecular Medicine, University Medical Center Utrecht, 3508 AB Utrecht, The Netherlands. 2. Nutrition, Metabolism & Genomics Group, Division of Human Nutrition, Wageningen University, 6703 HD Wageningen, The Netherlands. 3. Department of Pediatric Gastroenterology, Wilhelmina Children's Hospital, University Medical Center Utrecht, 3508 AB Utrecht, The Netherlands. 4. Department of Surgery, Rijnstate Hospital, 6815 AD Arnhem, The Netherlands. 5. Department of Molecular Cancer Research, Center for Molecular Medicine, University Medical Center Utrecht, 3508 AB Utrecht, The Netherlands. Electronic address: S.W.C.vanmil@umcutrecht.nl.
Abstract
BACKGROUND & AIMS: The bile acid-activated farnesoid X receptor (FXR) is a nuclear receptor regulating bile acid, glucose and cholesterol homeostasis. Obeticholic acid (OCA), a promising drug for the treatment of non-alcoholic steatohepatitis (NASH) and type 2 diabetes, activates FXR. Mouse studies demonstrated that FXR activation by OCA alters hepatic expression of many genes. However, no data are available on the effects of OCA in the human liver. Here we generated gene expression profiles in human precision cut liver slices (hPCLS) after treatment with OCA. METHODS: hPCLS were incubated with OCA for 24 h. Wild-type or FXR(-/-) mice received OCA or vehicle by oral gavage for 7 days. RESULTS: Transcriptomic analysis showed that well-known FXR target genes, including NR0B2 (SHP), ABCB11 (BSEP), SLC51A (OSTα) and SLC51B (OSTβ), and ABCB4 (MDR3) are regulated by OCA in hPCLS. Ingenuity pathway analysis confirmed that 'FXR/RXR activation' is the most significantly changed pathway upon OCA treatment. Comparison of gene expression profiles in hPCLS and mouse livers identified 18 common potential FXR targets. ChIP-sequencing in mouse liver confirmed FXR binding to IR1 sequences of Akap13, Cgnl1, Dyrk3, Pdia5, Ppp1r3b and Tbx6. CONCLUSIONS: Our study shows that hPCLS respond to OCA treatment by upregulating well-known FXR target genes, demonstrating its suitability to study FXR-mediated gene regulation. We identified six novel bona-fide FXR target genes in both mouse and human liver. Finally, we discuss a possible explanation for changes in high or low density lipoprotein observed in NASH and primary biliary cholangitis patients treated with OCA based on the genomic expression profile in hPCLS.
BACKGROUND & AIMS: The bile acid-activated farnesoid X receptor (FXR) is a nuclear receptor regulating bile acid, glucose and cholesterol homeostasis. Obeticholic acid (OCA), a promising drug for the treatment of non-alcoholic steatohepatitis (NASH) and type 2 diabetes, activates FXR. Mouse studies demonstrated that FXR activation by OCA alters hepatic expression of many genes. However, no data are available on the effects of OCA in the human liver. Here we generated gene expression profiles in human precision cut liver slices (hPCLS) after treatment with OCA. METHODS: hPCLS were incubated with OCA for 24 h. Wild-type or FXR(-/-) mice received OCA or vehicle by oral gavage for 7 days. RESULTS: Transcriptomic analysis showed that well-known FXR target genes, including NR0B2 (SHP), ABCB11 (BSEP), SLC51A (OSTα) and SLC51B (OSTβ), and ABCB4 (MDR3) are regulated by OCA in hPCLS. Ingenuity pathway analysis confirmed that 'FXR/RXR activation' is the most significantly changed pathway upon OCA treatment. Comparison of gene expression profiles in hPCLS and mouse livers identified 18 common potential FXR targets. ChIP-sequencing in mouse liver confirmed FXR binding to IR1 sequences of Akap13, Cgnl1, Dyrk3, Pdia5, Ppp1r3b and Tbx6. CONCLUSIONS: Our study shows that hPCLS respond to OCA treatment by upregulating well-known FXR target genes, demonstrating its suitability to study FXR-mediated gene regulation. We identified six novel bona-fide FXR target genes in both mouse and human liver. Finally, we discuss a possible explanation for changes in high or low density lipoprotein observed in NASH and primary biliary cholangitispatients treated with OCA based on the genomic expression profile in hPCLS.
Authors: Xia Wu; Jessica B Roberto; Allison Knupp; Heidi L Kenerson; Camtu D Truong; Sebastian Y Yuen; Katherine J Brempelis; Marianne Tuefferd; Antony Chen; Helen Horton; Raymond S Yeung; Ian N Crispe Journal: J Immunol Methods Date: 2018-02-01 Impact factor: 2.303
Authors: A Dash; R A Figler; B R Blackman; S Marukian; M S Collado; M J Lawson; S A Hoang; A J Mackey; D Manka; B K Cole; R E Feaver; A J Sanyal; B R Wamhoff Journal: Toxicol In Vitro Date: 2016-12-07 Impact factor: 3.500