PMID- 32761972 OWN - NLM STAT- MEDLINE DCOM- 20220107 LR - 20220716 IS - 1527-3350 (Electronic) IS - 0270-9139 (Print) IS - 0270-9139 (Linking) VI - 73 IP - 6 DP - 2021 Jun TI - Mast Cells Induce Ductular Reaction Mimicking Liver Injury in Mice Through Mast Cell-Derived Transforming Growth Factor Beta 1 Signaling. PG - 2397-2410 LID - 10.1002/hep.31497 [doi] AB - BACKGROUND AND AIMS: Following liver injury, mast cells (MCs) migrate into the liver and are activated in patients with cholestasis. Inhibition of MC mediators decreases ductular reaction (DR) and liver fibrosis. Transforming growth factor beta 1 (TGF-beta1) contributes to fibrosis and promotes liver disease. Our aim was to demonstrate that reintroduction of MCs induces cholestatic injury through TGF-beta1. APPROACH AND RESULTS: Wild-type, Kit(W-sh) (MC-deficient), and multidrug resistance transporter 2/ABC transporter B family member 2 knockout mice lacking l-histidine decarboxylase were injected with vehicle or PKH26-tagged murine MCs pretreated with 0.01% dimethyl sulfoxide (DMSO) or the TGF-beta1 receptor inhibitor (TGF-betaRi), LY2109761 (10 muM) 3 days before sacrifice. Hepatic damage was assessed by hematoxylin and eosin (H&E) and serum chemistry. Injected MCs were detected in liver, spleen, and lung by immunofluorescence (IF). DR was measured by cytokeratin 19 (CK-19) immunohistochemistry and F4/80 staining coupled with real-time quantitative PCR (qPCR) for interleukin (IL)-1beta, IL-33, and F4/80; biliary senescence was evaluated by IF or qPCR for p16, p18, and p21. Fibrosis was evaluated by sirius red/fast green staining and IF for synaptophysin 9 (SYP-9), desmin, and alpha smooth muscle actin (alpha-SMA). TGF-beta1 secretion/expression was measured by enzyme immunoassay and qPCR. Angiogenesis was detected by IF for von Willebrand factor and vascular endothelial growth factor C qPCR. In vitro, MC-TGF-beta1 expression/secretion were measured after TGF-betaRi treatment; conditioned medium was collected. Cholangiocytes and hepatic stellate cells (HSCs) were treated with MC-conditioned medium, and biliary proliferation/senescence was measured by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium and qPCR; HSC activation evaluated for alpha-SMA, SYP-9, and collagen type-1a expression. MC injection recapitulates cholestatic liver injury characterized by increased DR, fibrosis/TGF-beta1 secretion, and angiogenesis. Injection of MC-TGF-betaRi reversed these parameters. In vitro, MCs induce biliary proliferation/senescence and HSC activation that was reversed with MCs lacking TGF-beta1. CONCLUSIONS: Our study demonstrates that reintroduction of MCs mimics cholestatic liver injury and that MC-derived TGF-beta1 may be a target in chronic cholestatic liver disease. CI - (c) 2020 by the American Association for the Study of Liver Diseases. FAU - Kyritsi, Konstantina AU - Kyritsi K AD - Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine Research, Indianapolis, IN. FAU - Kennedy, Lindsey AU - Kennedy L AD - Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine Research, Indianapolis, IN. FAU - Meadows, Vik AU - Meadows V AD - Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine Research, Indianapolis, IN. AD - Richard L. Roudebush VA Medical Center, Indiana University School of Medicine Research, Indianapolis, IN. FAU - Hargrove, Laura AU - Hargrove L AD - Texas A&M University Health Science Center, Temple, TX. FAU - Demieville, Jennifer AU - Demieville J AD - Central Texas Veterans Health Care System, Temple, TX. FAU - Pham, Linh AU - Pham L AD - Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine Research, Indianapolis, IN. FAU - Sybenga, Amelia AU - Sybenga A AD - UVM Health Network, Burlington, VT. FAU - Kundu, Debjyoti AU - Kundu D AD - Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine Research, Indianapolis, IN. FAU - Cerritos, Karla AU - Cerritos K AD - Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine Research, Indianapolis, IN. FAU - Meng, Fanyin AU - Meng F AD - Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine Research, Indianapolis, IN. AD - Richard L. Roudebush VA Medical Center, Indiana University School of Medicine Research, Indianapolis, IN. FAU - Alpini, Gianfranco AU - Alpini G AD - Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine Research, Indianapolis, IN. AD - Richard L. Roudebush VA Medical Center, Indiana University School of Medicine Research, Indianapolis, IN. FAU - Francis, Heather AU - Francis H AD - Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine Research, Indianapolis, IN. AD - Richard L. Roudebush VA Medical Center, Indiana University School of Medicine Research, Indianapolis, IN. LA - eng GR - R01 DK108959/DK/NIDDK NIH HHS/United States GR - I01 BX000574/BX/BLRD VA/United States GR - I01 BX003031/BX/BLRD VA/United States GR - IK6 BX004601/BX/BLRD VA/United States GR - R01 DK115184/DK/NIDDK NIH HHS/United States GR - R01 DK119421/DK/NIDDK NIH HHS/United States GR - IK6 BX005226/BX/BLRD VA/United States GR - I01 BX001724/BX/BLRD VA/United States GR - R01 DK076898/DK/NIDDK NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20210419 PL - United States TA - Hepatology JT - Hepatology (Baltimore, Md.) JID - 8302946 RN - 0 (Acta2 protein, mouse) RN - 0 (Actins) RN - 0 (Transforming Growth Factor beta1) RN - 0 (Vascular Endothelial Growth Factor C) RN - 0 (vascular endothelial growth factor C, mouse) RN - 820484N8I3 (Histamine) SB - IM MH - Actins/*metabolism MH - Animals MH - Bile Ducts/metabolism/pathology MH - Cell Migration Assays MH - Cell Proliferation MH - Cellular Senescence MH - Cholestasis, Intrahepatic/*metabolism MH - Drug Discovery MH - Hepatic Stellate Cells MH - Histamine/blood MH - Liver/*pathology MH - *Liver Cirrhosis/drug therapy/metabolism MH - *Mast Cells/metabolism/pathology MH - Mice MH - Signal Transduction MH - *Transforming Growth Factor beta1/antagonists & inhibitors/metabolism MH - Up-Regulation MH - Vascular Endothelial Growth Factor C/*metabolism PMC - PMC7864988 MID - NIHMS1630031 EDAT- 2020/08/08 06:00 MHDA- 2022/01/08 06:00 PMCR- 2022/06/01 CRDT- 2020/08/08 06:00 PHST- 2020/06/15 00:00 [revised] PHST- 2020/04/03 00:00 [received] PHST- 2020/06/28 00:00 [accepted] PHST- 2020/08/08 06:00 [pubmed] PHST- 2022/01/08 06:00 [medline] PHST- 2020/08/08 06:00 [entrez] PHST- 2022/06/01 00:00 [pmc-release] AID - 10.1002/hep.31497 [doi] PST - ppublish SO - Hepatology. 2021 Jun;73(6):2397-2410. doi: 10.1002/hep.31497. Epub 2021 Apr 19.