233 results on '"Julie Venter"'
Search Results
2. Supplementary Figure 5 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
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Sharon DeMorrow, Antonio Benedetti, Marco Marzioni, Sum P. Lee, Domenico Alvaro, Gabriel Frampton, Monique Coufal, Antonio Franchitto, Paolo Onori, Francesca Bernuzzi, Shelley Kopriva, Julie Venter, Eugenio Gaudio, Pietro Invernizzi, and Gianfranco Alpini
- Abstract
Supplementary Figure 5 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
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- 2023
3. Supplementary Figure 1 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
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Sharon DeMorrow, Antonio Benedetti, Marco Marzioni, Sum P. Lee, Domenico Alvaro, Gabriel Frampton, Monique Coufal, Antonio Franchitto, Paolo Onori, Francesca Bernuzzi, Shelley Kopriva, Julie Venter, Eugenio Gaudio, Pietro Invernizzi, and Gianfranco Alpini
- Abstract
Supplementary Figure 1 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
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- 2023
4. Supplementary Figure 2 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
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Sharon DeMorrow, Antonio Benedetti, Marco Marzioni, Sum P. Lee, Domenico Alvaro, Gabriel Frampton, Monique Coufal, Antonio Franchitto, Paolo Onori, Francesca Bernuzzi, Shelley Kopriva, Julie Venter, Eugenio Gaudio, Pietro Invernizzi, and Gianfranco Alpini
- Abstract
Supplementary Figure 2 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
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- 2023
5. Supplementary Figure Legends 1-5 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
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Sharon DeMorrow, Antonio Benedetti, Marco Marzioni, Sum P. Lee, Domenico Alvaro, Gabriel Frampton, Monique Coufal, Antonio Franchitto, Paolo Onori, Francesca Bernuzzi, Shelley Kopriva, Julie Venter, Eugenio Gaudio, Pietro Invernizzi, and Gianfranco Alpini
- Abstract
Supplementary Figure Legends 1-5 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
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- 2023
6. Supplementary Figure 3 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
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Sharon DeMorrow, Antonio Benedetti, Marco Marzioni, Sum P. Lee, Domenico Alvaro, Gabriel Frampton, Monique Coufal, Antonio Franchitto, Paolo Onori, Francesca Bernuzzi, Shelley Kopriva, Julie Venter, Eugenio Gaudio, Pietro Invernizzi, and Gianfranco Alpini
- Abstract
Supplementary Figure 3 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
- Published
- 2023
7. Supplementary Figure 4 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
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Sharon DeMorrow, Antonio Benedetti, Marco Marzioni, Sum P. Lee, Domenico Alvaro, Gabriel Frampton, Monique Coufal, Antonio Franchitto, Paolo Onori, Francesca Bernuzzi, Shelley Kopriva, Julie Venter, Eugenio Gaudio, Pietro Invernizzi, and Gianfranco Alpini
- Abstract
Supplementary Figure 4 from Serotonin Metabolism Is Dysregulated in Cholangiocarcinoma, which Has Implications for Tumor Growth
- Published
- 2023
8. Leptin Enhances Hepatic Fibrosis and Inflammation in a Mouse Model of Cholestasis
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Anca D. Petrescu, Stephanie Grant, Elaina Williams, Su Yeon An, Nikhil Seth, Mark Shell, Tyson Amundsen, Christopher Tan, Yusra Nadeem, Matthew Tjahja, Lancaster Weld, Christopher S. Chu, Julie Venter, Gabriel Frampton, Matthew McMillin, and Sharon DeMorrow
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Inflammation ,Leptin ,Liver Cirrhosis ,Male ,Mice, Knockout ,Cholestasis ,Hyperplasia ,Regular Article ,Antibodies, Neutralizing ,Pathology and Forensic Medicine ,Disease Models, Animal ,Mice ,Liver ,Hepatic Stellate Cells ,Animals ,Cytokines ,Receptors, Leptin ,Female ,Proto-Oncogene Proteins c-akt - Abstract
Leptin is an adipokine with roles in food intake and energy metabolism through its actions on neurons in the hypothalamus. The role of leptin in obesity and cardiovascular disorders is well documented. However, its influence on liver conditions such as cholestasis is poorly understood. The effects of exogenous leptin and leptin-neutralizing antibody on biliary hyperplasia, hepatic fibrosis, and inflammation in the multidrug resistance protein 2 knockout (Mdr2KO) mouse model of cholestasis were assessed by quantifying markers specific for cholangiocytes, activated hepatic stellate cells (HSCs), and cytokines. Serum and hepatic leptin were increased in Mdr2KO mice compared with FVB/NJ (FVBN) controls, and exogenous leptin enhanced biliary hyperplasia and liver fibrosis in Mdr2KO and FVBN mice. Leptin administration increased hepatic expression of C-C motif chemokine ligand 2 and IL-6 in Mdr2KO mice. In contrast, leptin-neutralizing antibody reduced intrahepatic bile duct mass and decreased HSC activation in Mdr2KO mice compared with FVBN controls. Sex-related differences were noted, with female Mdr2KO mice having more leptin than males. In cholangiocytes and LX2 cells in vitro, leptin increased phosphorylated Akt and stimulated cell proliferation. Leptin receptor siRNA and inhibitors of Akt phosphorylation impaired leptin-induced cell proliferation and proinflammatory cytokines. The current data suggest that leptin is abnormally increased in cholestatic mice, and excess leptin increases ductular reaction, hepatic fibrosis, and inflammation via leptin receptor–mediated phosphorylation of Akt in cholangiocytes and HSCs.
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- 2022
9. Targeting Galanin Receptor Signaling as a Novel Therapeutic Strategy for the Treatment of Fatty Liver Disease
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Sharon DeMorrow, Elaina Williams, Su Yeon An, Julie Venter, Won Seog Choi, Yusra Nadeem, Christopher Chu, and Anca Petrescu
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Genetics ,Molecular Biology ,Biochemistry ,Biotechnology - Published
- 2022
10. Knockout of α-calcitonin gene-related peptide attenuates cholestatic liver injury by differentially regulating cellular senescence of hepatic stellate cells and cholangiocytes
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Tianhao Zhou, Lixian Chen, Paul Baker, Konstantina Kyritsi, Ying Wan, Amelia Sybenga, Shannon Glaser, Gianfranco Alpini, Nan Wu, Chaodong Wu, Fanyin Meng, Heather Francis, Pietro Invernizzi, Julie Venter, Francesca Bernuzzi, Qiaobing Huang, Ludovica Ceci, Wan, Y, Ceci, L, Wu, N, Zhou, T, Chen, L, Venter, J, Francis, H, Bernuzzi, F, Invernizzi, P, Kyritsi, K, Baker, P, Huang, Q, Wu, C, Sybenga, A, Alpini, G, Meng, F, and Glaser, S
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Liver Cirrhosis ,Male ,0301 basic medicine ,MAPK/ERK pathway ,Senescence ,MAP Kinase Signaling System ,Calcitonin Gene-Related Peptide ,cholestatic liver diseases ,Cholangitis, Sclerosing ,Cholangiocyte proliferation ,Calcitonin gene-related peptide ,Article ,Cholangiocyte ,Pathology and Forensic Medicine ,03 medical and health sciences ,biliary tract ,cellular senescence ,liver fibrosis ,primary sclerosing cholangitis ,0302 clinical medicine ,MED/12 - GASTROENTEROLOGIA ,Fibrosis ,Hepatic Stellate Cells ,medicine ,Animals ,Humans ,Molecular Biology ,Mice, Knockout ,Liver injury ,integumentary system ,Chemistry ,Cell Biology ,medicine.disease ,3. Good health ,Cell biology ,Mice, Inbred C57BL ,030104 developmental biology ,nervous system ,Case-Control Studies ,030220 oncology & carcinogenesis ,Hepatic stellate cell ,Female - Abstract
Background: α-Calcitonin gene-related peptide (α-CGRP) is a 37-amino acid neuropeptide involved in several pathophysiological processes. α-CGRP is involved in the regulation of cholangiocyte proliferation during cholestasis. In this study, we aimed to evaluate if α-CGRP regulates bile duct ligation (BDL)-induced liver fibrosis by using a α-CGRP knockout (α-CGRP−/−) mouse model. Methods: α-CGRP−/− and wild-type (WT) mice were subjected to sham surgery or BDL for 7 days. Then, liver fibrosis and cellular senescence as well as the expression of kinase such as p38 and C-Jun N-terminal protein kinase (JNK) in mitogen-activated protein kinases (MAPK) signaling pathway were evaluated in total liver, together with measurement of cellular senescence in cholangiocytes or hepatic stellate cells (HSCs). Results: There was enhanced hepatic expression of Calca (coding α-CGRP) and the CGRP-receptor components (CRLR, RAMP-1 and RCP) in BDL and in both WT α-CGRP−/− and BDL α-CGRP−/− mice, respectively. Moreover, there was increased CGRP serum levels and hepatic mRNA expression of CALCA and CGRP receptor components in late-stage PSC samples compared to healthy control samples. Depletion of α-CGRP reduced liver injury and fibrosis in BDL mice that was associated with enhanced cellular senescence of hepatic stellate cells and reduced senescence of cholangiocytes as well as decreased activation of p38 and JNK MAPK signaling pathway. Cholangiocyte supernatant from BDL α-CGRP−/− mice inhibited the activation and increased cellular senescence of cultured human HSCs (HHSCs) compared to HHSCs stimulated with BDL cholangiocyte supernatant. Taken together, endogenous α-CGRP promoted BDL-induced cholestatic liver fibrosis through differential changes in senescence of HSCs and cholangiocytes and activation of p38 and JNK signaling. Modulation of α-CGRP/CGRP receptor signaling may be key for the management of biliary senescence and liver fibrosis in cholangiopathies.
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- 2019
11. Amelioration of Ductular Reaction by Stem Cell Derived Extracellular Vesicles in MDR2 Knockout Mice via Lethal‐7 microRNA
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Heather Francis, Shannon Glaser, Nan Wu, Demeng Chen, Tianhao Zhou, Sugeily Ramos-Lorenzo, Fanyin Meng, Pietro Invernizzi, Li Huang, Kelly McDaniel, Julie Venter, Gianfranco Alpini, Francesca Bernuzzi, Ludovica Ceci, Chaodong Wu, Keisaku Sato, Mcdaniel, K, Wu, N, Zhou, T, Huan, L, Sato, K, Venter, J, Ceci, L, Chen, D, Ramos-Lorenzo, S, Invernizzi, P, Bernuzzi, F, Wu, C, Francis, H, Glaser, S, Alpini, G, and Meng, F
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ductular reaction ,Liver Cirrhosis ,0301 basic medicine ,Cell signaling ,ATP Binding Cassette Transporter, Subfamily B ,liver stem cells ,Cholangitis, Sclerosing ,Liver Stem Cell ,Real-Time Polymerase Chain Reaction ,LIN28 ,Sensitivity and Specificity ,Article ,Cholangiocyte ,Mice ,Random Allocation ,03 medical and health sciences ,0302 clinical medicine ,MED/12 - GASTROENTEROLOGIA ,Risk Factors ,Fibrosis ,medicine ,Animals ,Humans ,Secretion ,Cells, Cultured ,Mice, Knockout ,Hepatology ,Chemistry ,Stem Cells ,liver fibrosi ,Cell Differentiation ,Primary sclerosing cholangiti ,medicine.disease ,Cell biology ,Disease Models, Animal ,MicroRNAs ,030104 developmental biology ,Hepatocytes ,Hepatic stellate cell ,Female ,030211 gastroenterology & hepatology ,extracellular vesicle ,Stem cell - Abstract
Cholangiopathies are diseases that affect cholangiocytes, the cells lining the biliary tract. Liver stem cells (LSCs) are able to differentiate into all cells of the liver and possibly influence the surrounding liver tissue by secretion of signaling molecules. One way in which cells can interact is through secretion of extracellular vesicles (EVs), which are small membrane-bound vesicles that contain proteins, microRNAs (miRNAs), and cytokines. We evaluated the contents of liver stem cell–derived EVs (LSCEVs), compared their miRNA contents to those of EVs isolated from hepatocytes, and evaluated the downstream targets of these miRNAs. We finally evaluated the crosstalk among LSCs, cholangiocytes, and human hepatic stellate cells (HSCs). We showed that LSCEVs were able to reduce ductular reaction and biliary fibrosis in multidrug resistance protein 2 (MDR2)(−/−) mice. Additionally, we showed that cholangiocyte growth was reduced and HSCs were deactivated in LSCEV-treated mice. Evaluation of LSCEV contents compared with EVs derived from hepatocytes showed a large increase in the miRNA, lethal-7 (let-7). Further evaluation of let-7 in MDR2(−/−) mice and human primary sclerosing cholangitis samples showed reduced levels of let-7 compared with controls. In liver tissues and isolated cholangiocytes, downstream targets of let-7 (identified by ingenuity pathway analysis), Lin28a (Lin28 homolog A), Lin28b (Lin28 homolog B), IL-13 (interleukin 13), NR1H4 (nuclear receptor subfamily 1 group H member 4) and NF-κB (nuclear factor kappa B), are elevated in MDR2(−/−) mice, but treatment with LSCEVs reduced levels of these mediators of ductular reaction and biliary fibrosis through the inhibition of NF-κB and IL-13 signaling pathways. Evaluation of crosstalk using cholangiocyte supernatants from LSCEV-treated cells on cultured HSCs showed that HSCs had reduced levels of fibrosis and increased senescence. CONCLUSION: Our studies indicate that LSCEVs could be a possible treatment for cholangiopathies or could be used for target validation for future therapies.
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- 2019
12. Bile Acid Interactions with Cholangiocytes
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Gianfranco Alpini, Shannon Glaser, Heather Francis, Marco Marzioni, Julie Venter, Jo Lynne Phinizy, and Gene LeSage
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- 2020
13. Modulation of the Tryptophan Hydroxylase 1/Monoamine Oxidase-A/5-Hydroxytryptamine/5-Hydroxytryptamine Receptor 2A/2B/2C Axis Regulates Biliary Proliferation and Liver Fibrosis During Cholestasis
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David C. Zawieja, Romil Saxena, Tianhao Zhou, Luca Fabris, Heather Francis, Shannon Glaser, Lixian Chen, Nan Wu, Chaodong Wu, April O'Brien, Travis W. Hein, Nicholas J. Skill, Anatoliy A. Gashev, Ludovica Ceci, Fanyin Meng, Konstantina Kyritsi, Gianfranco Alpini, Suthat Liangpunsakul, Pietro Invernizzi, Julie Venter, Kyritsi, K, Chen, L, O'Brien, A, Francis, H, Hein, T, Venter, J, Wu, N, Ceci, L, Zhou, T, Zawieja, D, Gashev, A, Meng, F, Invernizzi, P, Fabris, L, Wu, C, Skill, N, Saxena, R, Liangpunsakul, S, Alpini, G, and Glaser, S
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0301 basic medicine ,Liver Cirrhosis ,Male ,medicine.medical_specialty ,Serotonin ,ATP Binding Cassette Transporter, Subfamily B ,Monoamine oxidase ,Cholangitis, Sclerosing ,Tryptophan Hydroxylase ,digestive system ,Article ,Proinflammatory cytokine ,Rats, Sprague-Dawley ,03 medical and health sciences ,Mice ,0302 clinical medicine ,MED/12 - GASTROENTEROLOGIA ,Internal medicine ,Receptor, Serotonin, 5-HT2B ,medicine ,Receptor, Serotonin, 5-HT2C ,Animals ,Humans ,Receptor, Serotonin, 5-HT2A ,cholangiocytes ,Receptor ,serotonin ,cholestasis ,Monoamine Oxidase ,Cell Proliferation ,TPH1 ,Hepatology ,Chemistry ,cholangiopathie ,Tryptophan hydroxylase ,Rats ,030104 developmental biology ,Endocrinology ,Receptors, Serotonin ,Hepatic stellate cell ,Enterochromaffin cell ,030211 gastroenterology & hepatology ,Bile Ducts ,fibrosi - Abstract
BACKGROUND AND AIMS: Serotonin (5HT) is a neuroendocrine hormone synthetized in the central nervous system (CNS) as well as enterochromaffin cells of the gastrointestinal tract. Tryptophan hydroxylase (TPH1) and monoamine oxidase (MAO-A) are the key enzymes for the synthesis and catabolism of 5HT, respectively. Previous studies demonstrated that 5-hydroxytryptamine receptor (5HTR)1A/1B receptor agonists inhibit biliary hyperplasia in bile-duct ligated (BDL) rats, whereas 5HTR2B receptor antagonists attenuate liver fibrosis (LF) in mice. Our aim was to evaluate the role of 5HTR2A/2B/2C agonists/antagonists in cholestatic models. APPROACH AND RESULTS: While in vivo studies were performed in BDL rats and the multidrug resistance gene 2 knockout (Mdr2(−/−)) mouse model of PSC, in vitro studies were performed in cell lines of cholangiocytes and hepatic stellate cells (HSCs). 5HTR2A/2B/2C and MAO-A/TPH1 are expressed in cholangiocytes and HSCs from BDL rats and Mdr2(−/−) mice. Ductular reaction, LF, as well as the mRNA expression of proinflammatory genes increased in normal, BDL rats, and Mdr2(−/−) mice following treatment 5HTR2A/2B/2C agonists, but decreased when BDL rats and Mdr2(−/−) mice were treated with 5HTR2A/2B/2C antagonists compared to BDL rats and Mdr2(−/−) mice, respectively. 5HT levels increase in Mdr2(−/−) mice and in PSC human patients compared to their controls and decrease in serum of Mdr2(−/−) mice treated with 5HTR2A/2B/2C antagonists compared to untreated Mdr2(−/−) mice. In vitro, cell lines of murine cholangiocytes and human HSCs express 5HTR2A/2B/2C and MAO-A/TPH1; treatment of these cell lines with 5HTR2A/2B/2C antagonists or TPH1 inhibitor decreased 5HT levels as well as expression of fibrosis and inflammation genes compared to controls. CONCLUSIONS: Modulation of the TPH1/MAO-A/5HT/5HTR2A/2B/2C axis may represent a therapeutic approach for management of cholangiopathies, including PSC.
- Published
- 2020
14. α7-nAChR Knockout Mice Decreases Biliary Hyperplasia and Liver Fibrosis in Cholestatic Bile Duct-Ligated Mice
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Tori White, Nan Wu, Laurent Ehrlich, Marinda Scrushy, David E. Dostal, Chad Hall, Gianfranco Alpini, Shannon Glaser, Terry C. Lairmore, Julie Venter, Fanyin Meng, Chaodong Wu, Muhammad Mubarak, April O'Brien, and Lixian Chen
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Liver Cirrhosis ,Male ,0301 basic medicine ,Pathology ,medicine.medical_specialty ,alpha7 Nicotinic Acetylcholine Receptor ,Intrahepatic bile ducts ,digestive system ,Article ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cholestasis ,Transforming Growth Factor beta ,Fibrosis ,Cell Line, Tumor ,Proliferating Cell Nuclear Antigen ,Genetics ,medicine ,Animals ,Humans ,Molecular Biology ,Sirius Red ,Hyperplasia ,Biliary hyperplasia ,Bile duct ,business.industry ,Kupffer cell ,Cholestasis, Extrahepatic ,medicine.disease ,Mice, Inbred C57BL ,Ki-67 Antigen ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Hepatic stellate cell ,Cytokines ,030211 gastroenterology & hepatology ,Bile Ducts ,business - Abstract
α7-nAChR is a nicotinic acetylcholine receptor [specifically expressed on hepatic stellate cells (HSCs), Kupffer cells, and cholangiocytes] that regulates inflammation and apoptosis in the liver. Thus, targeting α7-nAChR may be therapeutic in biliary diseases. Bile duct ligation (BDL) was performed on wild-type (WT) and α7-nAChR−/− mice. We first evaluated the expression of α7-nAChR by immunohistochemistry (IHC) in liver sections. IHC was also performed to assess intrahepatic bile duct mass (IBDM), and Sirius Red staining was performed to quantify the amount of collagen deposition. Immunofluorescence was performed to assess colocalization of α7-nAChR with bile ducts (costained with CK-19) and HSCs (costained with desmin). The mRNA expression of α7-nAChR, Ki-67/PCNA (proliferation), fibrosis genes (TGF-β1, fibronectin-1, Col1α1, and α-SMA), and inflammatory markers (IL-6, IL-1β, and TNF-α) was measured by real-time PCR. Biliary TGF-β1 and hepatic CD68 (Kupffer cell marker) expression was assessed using IHC. α7-nAChR immunoreactivity was observed in both bile ducts and HSCs and increased following BDL. α7-nAChR−/− BDL mice exhibited decreased (i) bile duct mass, liver fibrosis, and inflammation, and (ii) immunoreactivity of TGF-β1 as well as expression of fibrosis genes compared to WT BDL mice. α7-nAChR activation triggers biliary proliferation and liver fibrosis and may be a therapeutic target in managing extrahepatic biliary obstruction.
- Published
- 2018
15. Prolonged darkness reduces liver fibrosis in a mouse model of primary sclerosing cholangitis by miR‐200b down‐regulation
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Fanyin Meng, Heather Francis, Eugenio Gaudio, Gianfranco Alpini, Sharon DeMorrow, Yuyan Han, Lindsey Kennedy, Shannon Glaser, Nan Wu, Romina Mancinelli, Tianhao Zhou, Paolo Onori, Francesca Bernuzzi, Pietro Invernizzi, Julie Venter, Antonio Franchitto, Wu, N, Meng, F, Zhou, T, Han, Y, Kennedy, L, Venter, J, Francis, H, Demorrow, S, Onori, P, Invernizzi, P, Bernuzzi, F, Mancinelli, R, Gaudio, E, Franchitto, A, Glaser, S, and Alpini, G
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Liver Cirrhosis ,Biliary epithelium ,Cholangiopathy ,Male ,0301 basic medicine ,Pathology ,Fibrosi ,Angiogenesis ,AANAT ,Biochemistry ,angiogenesis ,Fibrosis ,Melatonin ,Biliary hyperplasia ,MicroRNA ,Darkness ,biliary epithelium ,cholangiopathy ,cholestasis ,miRNA ,Angiogenesi ,Cholestasi ,Biotechnology ,medicine.drug ,medicine.medical_specialty ,Liver Cirrhosi ,Cholangitis, Sclerosing ,Angiogenesis Inducing Agent ,Down-Regulation ,Mice, Transgenic ,Primary sclerosing cholangitis ,03 medical and health sciences ,Internal medicine ,Hepatic Stellate Cells ,Genetics ,medicine ,Animals ,Molecular Biology ,Cell Proliferation ,Animal ,business.industry ,Research ,medicine.disease ,Hepatic Stellate Cell ,MicroRNAs ,Disease Models, Animal ,030104 developmental biology ,Endocrinology ,Hepatic stellate cell ,Angiogenesis Inducing Agents ,Darkne ,business ,Hepatic fibrosis - Abstract
Melatonin therapy or prolonged exposure to complete darkness reduces biliary hyperplasia and liver fibrosis in bile-duct–ligated (BDL) rats; however, no information exists in primary sclerosing cholangitis (PSC). Thus, we aimed to determine the therapeutic effects of prolonged dark therapy or melatonin administration on hepatic fibrosis in the multidrug resistance gene 2–knockout (Mdr2−/−) mouse model of PSC. Melatonin levels, biliary mass, liver fibrosis, angiogenesis and miR-200b expression were evaluated in wild-type and Mdr2−/− mice exposed to darkness or melatonin treatment or in male patients with PSC and healthy controls. Mdr2−/− mice were also treated with miR-200b inhibitor or control before evaluating biliary mass, liver fibrosis, and angiogenesis. After overexpression of arylalkylamine N-acetyltransferase (AANAT; the enzyme regulating melatonin synthesis) or inhibition of miR-200b in cholangiocytes and hepatic stellate cells in vitro, we evaluated angiogenesis and fibrosis gene expression. After exposure to darkness or administration of melatonin, Mdr2−/− mice show elevated serum melatonin levels and inhibition of biliary mass, along with reduction of liver fibrosis and angiogenesis. MicroRNA PCR analysis demonstrated that miR-200b expression increased in Mdr2−/− mice and patients with PSC compared with controls and decreased in Mdr2−/− mice subjected to dark exposure or melatonin treatment. Inhibition of miR-200b in Mdr2−/− ablates biliary proliferation, liver fibrosis, and angiogenesis. In vitro, overexpression of AANAT or inhibition of miR-200b in cholangiocytes and hepatic stellate cells decreased the expression of miR-200b, angiogenesis, and fibrosis genes. Dark therapy or targeting melatonin/miR-200b axis may be important in the management of biliary damage and liver fibrosis in cholangiopathies including PSC.—Wu, N., Meng, F., Zhou, T., Han, Y., Kennedy, L., Venter, J., Francis, H., DeMorrow, S., Onori, P., Invernizzi, P., Bernuzzi, F., Mancinelli, R., Gaudio, E., Franchitto, A., Glaser, S., Alpini G. Prolonged darkness reduces liver fibrosis in a mouse model of primary sclerosing cholangitis by miR-200b down-regulation.
- Published
- 2017
16. Knockdown of Hepatic Gonadotropin-Releasing Hormone by Vivo-Morpholino Decreases Liver Fibrosis in Multidrug Resistance Gene 2 Knockout Mice by Down-Regulation of miR-200b
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Shannon Glaser, Paolo Onori, Antonio Franchitto, Julie Venter, Fanyin Meng, Kelly McDaniel, Lindsey Kennedy, Gianfranco Alpini, Romina Mancinelli, Konstantina Kyritsi, Francesca Bernuzzi, Tianhao Zhou, Nan Wu, Heather Francis, Domenico Alvaro, Eugenio Gaudio, Pietro Invernizzi, Kyritsi, K, Meng, F, Zhou, T, Wu, N, Venter, J, Francis, H, Kennedy, L, Onori, P, Franchitto, A, Bernuzzi, F, Invernizzi, P, Mcdaniel, K, Mancinelli, R, Alvaro, D, Gaudio, E, Alpini, G, and Glaser, S
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Liver Cirrhosis ,Male ,0301 basic medicine ,Gonadotropin-releasing hormone ,Morpholino ,liver ,cholangiocytes ,hepatic gonadotropin-releasing hormone ,Morpholinos ,Gonadotropin-Releasing Hormone ,Mice ,Fibrosis ,Mice, Knockout ,Gene knockdown ,Cholestasis ,GNRHR ,MicroRNA ,Regular Article ,Up-Regulation ,Liver ,Cholestasi ,Knockout mouse ,Disease Progression ,Human ,medicine.medical_specialty ,ATP Binding Cassette Transporter, Subfamily B ,Liver Cirrhosi ,Down-Regulation ,Biology ,Cholangiocyte ,Cell Line ,Pathology and Forensic Medicine ,03 medical and health sciences ,Internal medicine ,Hepatic Stellate Cells ,medicine ,Animals ,Humans ,Cell Proliferation ,Animal ,P-Glycoprotein ,medicine.disease ,Hepatic Stellate Cell ,Disease Models, Animal ,MicroRNAs ,030104 developmental biology ,Endocrinology ,Gene Expression Regulation ,Hepatic stellate cell ,Hepatic fibrosis ,Receptors, LHRH - Abstract
Hepatic fibrosis occurs during the progression of primary sclerosing cholangitis (PSC) and is characterized by accumulation of extracellular matrix proteins. Proliferating cholangiocytes and activated hepatic stellate cells (HSCs) participate in the promotion of liver fibrosis during cholestasis. Gonadotropin-releasing hormone (GnRH) is a trophic peptide hormone synthesized by hypothalamic neurons and the biliary epithelium and exerts its biological effects on cholangiocytes by interaction with the receptor subtype (GnRHR 1 ) expressed by cholangiocytes and HSCs. Previously, we demonstrated that administration of GnRH to normal rats increased intrahepatic biliary mass (IBDM) and hepatic fibrosis. Also, miR-200b is associated with the progression of hepatic fibrosis; however, the role of the GnRH/GnRHR 1 /miR-200b axis in the development of hepatic fibrosis in PSC is unknown. Herein, using the mouse model of PSC (multidrug resistance gene 2 knockout), the hepatic knockdown of GnRH decreased IBDM and liver fibrosis. In vivo and in vitro administration of GnRH increased the expression of miR-200b and fibrosis markers. The GnRH/GnRHR 1 axis and miR-200b were up-regulated in human PSC samples. Cetrorelix, a GnRHR 1 antagonist, inhibited the expression of fibrotic genes in vitro and decreased IBDM and hepatic fibrosis in vivo . Inhibition of miR-200b decreased the expression of fibrosis genes in vitro in cholangiocyte and HSC lines. Targeting the GnRH/GnRHR 1 /miR-200b axis may be key for the management of hepatic fibrosis during the progression of PSC.
- Published
- 2017
17. Substance P increases liver fibrosis by differential changes in senescence of cholangiocytes and hepatic stellate cells
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Nan Wu, Fanyin Meng, Heather Francis, Ying Wan, Lindsey Kennedy, Francesca Bernuzzi, Pietro Invernizzi, Julie Venter, Tianhao Zhou, Trenton Glaser, Shannon Glaser, Gianfranco Alpini, Qiaobing Huang, Wan, Y, Meng, F, Wu, N, Zhou, T, Venter, J, Francis, H, Kennedy, L, Glaser, T, Bernuzzi, F, Invernizzi, P, Glaser, S, Huang, Q, and Alpini, G
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Liver Cirrhosis ,Male ,0301 basic medicine ,Aging ,Apoptosis ,Substance P ,Mice ,Random Allocation ,chemistry.chemical_compound ,0302 clinical medicine ,Liver Function Tests ,Fibrosis ,Cells, Cultured ,Mice, Knockout ,Liver injury ,medicine.diagnostic_test ,Liver Function Test ,Biopsy, Needle ,Bile Duct ,Immunohistochemistry ,030220 oncology & carcinogenesis ,Human ,Senescence ,medicine.medical_specialty ,Liver Cirrhosi ,Biology ,Sensitivity and Specificity ,Cholangiocyte ,Primary sclerosing cholangitis ,03 medical and health sciences ,Internal medicine ,Hepatic Stellate Cells ,medicine ,Animals ,Humans ,RNA, Messenger ,Sirius Red ,Cell Proliferation ,Hepatology ,Animal ,Apoptosi ,Biomarker ,medicine.disease ,Hepatic Stellate Cell ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,Endocrinology ,chemistry ,Hepatic stellate cell ,Bile Ducts ,Liver function tests ,Biomarkers - Abstract
Substance P (SP) is involved in the proliferation of cholangiocytes in bile ductâligated (BDL) mice and human cholangiocarcinoma growth by interacting with the neurokinin-1 receptor (NK-1R). To identify whether SP regulates liver fibrosis during cholestasis, wild-type or NK-1R knockout (NK-1Râ/â) mice that received BDL or sham surgery and multidrug resistance protein 2 knockout (Mdr2â/â) mice treated with either an NK-1R antagonist (L-733,060) or saline were used. Additionally, wild-type mice were treated with SP or saline intraperitoneally. In vivo, there was increased expression of tachykinin precursor 1 (coding SP) and NK-1R in both BDL and Mdr2â/âmice compared to wild-type mice. Expression of tachykinin precursor 1 and NK-1R was significantly higher in liver samples from primary sclerosing cholangitis patients compared to healthy controls. Knockout of NK-1R decreased BDL-induced liver fibrosis, and treatment with L-733,060 resulted in decreased liver fibrosis in Mdr2â/âmice, which was shown by decreased sirius red staining, fibrosis gene and protein expression, and reduced transforming growth factor-β1 levels in serum and cholangiocyte supernatants. Furthermore, we observed that reduced liver fibrosis in NK-1Râ/âmice with BDL surgery or Mdr2â/âmice treated with L-733,060 was associated with enhanced cellular senescence of hepatic stellate cells and decreased senescence of cholangiocytes. In vitro, L-733,060 inhibited SP-induced expression of fibrotic genes in hepatic stellate cells and cholangiocytes; treatment with L-733,060 partially reversed the SP-induced decrease of senescence gene expression in cultured hepatic stellate cells and the SP-induced increase of senescence-related gene expression in cultured cholangiocytes. Conclusion: Collectively, our results demonstrate the regulatory effects of the SP/NK-1R axis on liver fibrosis through changes in cellular senescence during cholestatic liver injury. (Hepatology 2017;66:528â541)
- Published
- 2017
18. Inhibition of the apelin/apelin receptor axis decreases cholangiocarcinoma growth
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Tianhao Zhou, Francesca Bernuzzi, Shannon Glaser, Gianfranco Alpini, Chad Hall, Terry C. Lairmore, Pietro Invernizzi, Julie Venter, Tori White, April O'Brien, Fanyin Meng, Tien Dang, Laurent Ehrlich, Hall, C, Ehrlich, L, Venter, J, O'Brien, A, White, T, Zhou, T, Dang, T, Meng, F, Invernizzi, P, Bernuzzi, F, Alpini, G, Lairmore, T, and Glaser, S
- Subjects
Male ,Biliary epithelium ,0301 basic medicine ,MAPK/ERK pathway ,Cancer Research ,Angiogenesis ,Proliferation ,Angiogenesis Inhibitors ,Receptors, G-Protein-Coupled ,Cholangiocarcinoma ,Cell Movement ,Aged, 80 and over ,Apelin Receptors ,Mice, Inbred BALB C ,Neovascularization, Pathologic ,medicine.diagnostic_test ,Middle Aged ,Tumor Burden ,Apelin ,Vascular endothelial growth factor A ,Apelin receptor ,Oncology ,Vascular endothelial growth factor C ,Hypoxia-inducible factors ,cardiovascular system ,Intercellular Signaling Peptides and Proteins ,Female ,Signal Transduction ,Adult ,medicine.medical_specialty ,Mice, Nude ,Biology ,Article ,Flow cytometry ,03 medical and health sciences ,Cell Line, Tumor ,Internal medicine ,parasitic diseases ,medicine ,Animals ,Humans ,Neoplasm Invasiveness ,Cell Proliferation ,Pyrans ,fungi ,Xenograft Model Antitumor Assays ,030104 developmental biology ,Endocrinology ,Bile Duct Neoplasms ,Nitrobenzoates ,Cancer research - Abstract
Purpose Cholangiocarcinoma (CCA) is a malignancy of the biliary epithelium that is associated with low five-year survival. The apelin receptor (APLNR), which is activated by the apelin peptide, has not been studied in CCA. The purpose of this study is to determine if inhibition of the apelin/APLNR axis can inhibit CCA growth. Methods Immunohistochemistry, rtPCR, immunofluorescence, flow cytometry, and ELISA was used to measure APLNR expression in human CCA cells and tissues. Mz-ChA-1 cells were treated with increasing concentrations of apelin and ML221, an APLNR antagonist. Expression of proliferative and angiogenic genes were measured via rtPCR. In vivo , Mz-ChA-1 cells were injected into the flanks of nu/nu mice, which were treated with ML221 (150 μg/kg) via tail vein injection. Results Expression of the apelin/APLNR axis was increased in CCA. In vitro , CCA proliferation and angiogenesis was inhibited by ML221 treatment. ML221 treatment significantly decreased tumor growth in nu/nu mice. Conclusion The apelin/APLNR axis regulates CCA proliferation and angiogenesis. Inhibition of the apelin/APLNR axis decreases tumor growth in our xenograft model. Targeting APLNR signaling has the potential to serve as a novel, tumor directed therapy for CCA.
- Published
- 2017
19. Knockout of microRNA-21 reduces biliary hyperplasia and liver fibrosis in cholestatic bile duct ligated mice
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Tianhao Zhou, John F. Greene, Francesca Bernuzzi, Heather Francis, Walker Karstens, Nan Wu, Laura Hargrove, Shannon Glaser, Lindsey Kennedy, Konstantina Kyritsi, Gianfranco Alpini, Julie Venter, Pietro Invernizzi, Fanyin Meng, Kennedy, L, Meng, F, Venter, J, Zhou, T, Karstens, W, Hargrove, L, Wu, N, Kyritsi, K, Greene, J, Invernizzi, P, Bernuzzi, F, Glaser, S, Francis, H, and Alpini, G
- Subjects
Liver Cirrhosis ,Male ,0301 basic medicine ,Pathology ,medicine.medical_specialty ,Cirrhosis ,Apoptosis ,Cholestasis, Intrahepatic ,Article ,Cell Line ,Smad7 Protein ,Pathology and Forensic Medicine ,Mice ,03 medical and health sciences ,Cholestasis ,MED/12 - GASTROENTEROLOGIA ,Fibrosis ,Hepatic Stellate Cells ,medicine ,Animals ,Humans ,Molecular Biology ,Cells, Cultured ,Cell Proliferation ,Mice, Knockout ,Hyperplasia ,Bile duct ,Biliary hyperplasia ,business.industry ,Cell Biology ,medicine.disease ,Up-Regulation ,3. Good health ,Disease Models, Animal ,MicroRNAs ,Bile Ducts, Intrahepatic ,030104 developmental biology ,medicine.anatomical_structure ,Gene Expression Regulation ,Disease Progression ,Hepatic stellate cell ,RNA Interference ,Hepatic fibrosis ,business ,Biomarkers - Abstract
Cholestasis is a condition that leads to chronic hepatobiliary inflammation, fibrosis, and eventually cirrhosis. Many microRNAs (miRs) are known to have a role in fibrosis progression; however, the role of miR-21 during cholestasis remains unknown. Therefore, the aim of this study was to elucidate the role of miR-21 during cholestasis-induced biliary hyperplasia and hepatic fibrosis. Wild-type (WT) and miR-21-/- mice underwent Sham or bile duct ligation (BDL) for 1 week, before evaluating liver histology, biliary proliferation, hepatic stellate cell (HSC) activation, fibrotic response, and small mothers against decapentaplegic 7 (Smad-7) expression. In vitro, immortalized murine biliary cell lines (IMCLs) and human hepatic stellate cell line (hHSC) were treated with either miR-21 inhibitor or control before analyzing proliferation, apoptosis, and fibrotic responses. In vivo, the levels of miR-21 were increased in total liver and cholangiocytes after BDL, and loss of miR-21 decreased the amount of BDL-induced biliary proliferation and intrahepatic biliary mass. In addition, loss of miR-21 decreased BDL-induced HSC activation, collagen deposition, and expression of the fibrotic markers transforming growth factor-β1 and α-smooth muscle actin. In vitro, IMCL and hHSCs treated with miR-21 inhibitor displayed decreased proliferation and expression of fibrotic markers and enhanced apoptosis when compared with control treated cells. Furthermore, mice lacking miR-21 show increased Smad-7 expression, which may be driving the decrease in biliary hyperplasia and hepatic fibrosis. During cholestatic injury, miR-21 is increased and leads to increased biliary proliferation and hepatic fibrosis. Local modulation of miR-21 may be a therapeutic option for patients with cholestasis.
- Published
- 2016
20. Vasopressin regulates the growth of the biliary epithelium in polycystic liver disease
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Romina Mancinelli, Julie Venter, Luigi Pannarale, Paolo Onori, Shannon Glaser, Antonella Vetuschi, Francesca Olivero, Gianfranco Alpini, Guido Carpino, Antonio Franchitto, Eugenio Gaudio, and Roberta Sferra
- Subjects
Male ,0301 basic medicine ,Receptors, Vasopressin ,Vasopressin ,medicine.medical_specialty ,Vasopressins ,Neurohypophysial hormone ,Intrahepatic bile ducts ,Biology ,Epithelium ,Article ,Cell Line ,Pathology and Forensic Medicine ,03 medical and health sciences ,0302 clinical medicine ,Cholestasis ,Internal medicine ,Arginine vasopressin receptor 2 ,Cyclic AMP ,medicine ,Animals ,Humans ,Receptor ,Molecular Biology ,Keratin-19 ,Cysts ,Liver Diseases ,Polycystic liver disease ,Cell Biology ,medicine.disease ,Rats, Inbred F344 ,Mice, Inbred C57BL ,Bile Ducts, Intrahepatic ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,liver ,biliary tree ,vasopressin ,030211 gastroenterology & hepatology ,hormones, hormone substitutes, and hormone antagonists - Abstract
The neurohypophysial hormone arginine vasopressin (AVP) acts by three distinct receptor subtypes: V1a, V1b, and V2. In the liver, AVP is involved in ureogenesis, glycogenolysis, neoglucogenesis and regeneration. No data exist about the presence of AVP in the biliary epithelium. Cholangiocytes are the target cells in a number of animal models of cholestasis, including bile duct ligation (BDL), and in several human pathologies, such as polycystic liver disease characterized by the presence of cysts that bud from the biliary epithelium. In vivo, liver fragments from normal and BDL mice and rats as well as liver samples from normal and ADPKD patients were collected to evaluate: (i) intrahepatic bile duct mass by immunohistochemistry for cytokeratin-19; and (ii) expression of V1a, V1b and V2 by immunohistochemistry, immunofluorescence and real-time PCR. In vitro, small and large mouse cholangiocytes, H69 (non-malignant human cholangiocytes) and LCDE (human cholangiocytes from the cystic epithelium) were stimulated with vasopressin in the absence/presence of AVP antagonists such as OPC-31260 and Tolvaptan, before assessing cellular growth by MTT assay and cAMP levels. Cholangiocytes express V2 receptor that was upregulated following BDL and in ADPKD liver samples. Administration of AVP increased proliferation and cAMP levels of small cholangiocytes and LCDE cells. We found no effect in the proliferation of large mouse cholangiocytes and H69 cells. Increases were blocked by preincubation with the AVP antagonists. These results showed that AVP and its receptors may be important in the modulation of the proliferation rate of the biliary epithelium.
- Published
- 2016
21. Secretin/Secretin Receptor (Sct/SR) Signaling Promotes Biliary Senescence and Liver Fibrosis During Alcoholic Liver Disease
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Shannon Glaser, Fanyin Meng, Adrien Guillot, Tianhao Zhou, Nan Wu, Heather Francis, Bin Gao, Julie Venter, Ludovica Ceci, Gianfranco Alpini, Lindsey Kennedy, and Konstantina Kyritsi
- Subjects
Senescence ,Alcoholic liver disease ,medicine.medical_specialty ,business.industry ,Liver fibrosis ,medicine.disease ,Biochemistry ,Secretin ,Endocrinology ,Internal medicine ,Genetics ,medicine ,Secretin receptor ,business ,Molecular Biology ,Biotechnology - Published
- 2019
22. Secretin/secretin receptor signaling mediates biliary damage and liver fibrosis in early-stage primary biliary cholangitis
- Author
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Lindsey Kennedy, Nan Wu, Marco Marzioni, Julie Venter, Domenico Alvaro, M. Eric Gershwin, Fanyin Meng, Amelia Sybenga, Shannon Glaser, Paolo Onori, Gianfranco Alpini, Francesca Bernuzzi, Pietro Invernizzi, Marco Carbone, Luca Fabris, Heather Francis, Eugenio Gaudio, Antonio Franchitto, Kennedy, L, Francis, H, Invernizzi, P, Venter, J, Wu, N, Carbone, M, Eric Gershwin, M, Bernuzzi, F, Franchitto, A, Alvaro, D, Marzioni, M, Onori, P, Gaudio, E, Sybenga, A, Fabris, L, Meng, F, Glaser, S, and Alpini, G
- Subjects
0301 basic medicine ,Liver Cirrhosis ,Male ,Inbred C57BL ,Biochemistry ,angiogenesis ,cellular senescence ,cholangiopathies ,liver damage ,miR-125b ,Secretin ,Receptors, G-Protein-Coupled ,Mice ,0302 clinical medicine ,Receptor ,Mice, Knockout ,Gastrointestinal Hormone ,Liver Cirrhosis, Biliary ,Biliary ,cholangiopathie ,Gastrointestinal hormone ,Secretin receptor ,Female ,medicine.symptom ,Signal transduction ,Case-Control Studie ,Biliary Tract Disease ,Human ,Biotechnology ,Signal Transduction ,medicine.medical_specialty ,Liver Cirrhosi ,Knockout ,Biliary Tract Diseases ,Inflammation ,Receptors, Gastrointestinal Hormone ,Transforming Growth Factor beta1 ,G-Protein-Coupled ,03 medical and health sciences ,Internal medicine ,Genetics ,medicine ,Animals ,Humans ,Molecular Biology ,Animal ,business.industry ,Research ,Antagonist ,Receptor, Transforming Growth Factor-beta Type II ,angiogenesi ,digestive system diseases ,Mice, Inbred C57BL ,030104 developmental biology ,Endocrinology ,Case-Control Studies ,business ,030217 neurology & neurosurgery ,Transforming growth factor - Abstract
Primary biliary cholangitis (PBC) primarily targets cholangiocytes and is characterized by liver fibrosis and biliary proliferation. Activation of the secretin (Sct)/secretin receptor (SR) axis, expressed only by cholangiocytes, increases biliary proliferation, liver fibrosis, and bicarbonate secretion. We evaluated the effectiveness of SR antagonist treatment for early-stage PBC. Male and female dominant-negative TGF-β receptor II (dnTGF-βRII) (model of PBC) and wild-type mice at 12 wk of age were treated with saline or the SR antagonist, Sec 5-27, for 1 wk. dnTGF-βRII mice expressed features of early-stage PBC along with enhanced Sct/SR axis activation and Sct secretion. dnTGF-βRII mice had increased biliary proliferation or senescence, inflammation, and liver fibrosis. In dnTGF-βRII mice, there was increased microRNA-125b/TGF-β1/TGF-β receptor 1/VEGF-A signaling. Human early-stage PBC patients had an increase in hepatobiliary Sct and SR expression and serum Sct levels. Increased biliary Sct/SR signaling promotes biliary and hepatic damage during early-stage PBC.—Kennedy, L., Francis, H., Invernizzi, P., Venter, J., Wu, N., Carbone, M., Gershwin, M. E., Bernuzzi, F., Franchitto, A., Alvaro, D., Marzioni, M., Onori, P., Gaudio, E., Sybenga, A., Fabris, L., Meng, F., Glaser, S., Alpini, G. Secretin/secretin receptor signaling mediates biliary damage and liver fibrosis in early-stage primary biliary cholangitis. FASEB J. 33, 10269–10279 (2019). www.fasebj.org.
- Published
- 2019
23. Pinealectomy or light exposure exacerbates biliary damage and liver fibrosis in cholestatic rats through decreased melatonin synthesis
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April O'Brien, Linglin Xie, Fanyin Meng, Pietro Invernizzi, Nan Wu, Paolo Onori, Lixian Chen, Konstantina Kyritsi, Qiaobing Huang, David C. Zawieja, Eugenio Gaudio, Gianfranco Alpini, Ludovica Ceci, Tianhao Zhou, Chaodong Wu, Luca Fabris, Heather Francis, Amelia Sybenga, Julie Venter, Suthat Liangpunsakul, Shannon Glaser, Chen, L, Zhou, T, Wu, N, O'Brien, A, Venter, J, Ceci, L, Kyritsi, K, Onori, P, Gaudio, E, Sybenga, A, Xie, L, Wu, C, Fabris, L, Invernizzi, P, Zawieja, D, Liangpunsakul, S, Meng, F, Francis, H, Alpini, G, Huang, Q, and Glaser, S
- Subjects
Liver Cirrhosis ,Male ,Light ,AANAT ,medicine.medical_treatment ,CLOCK Proteins ,Pineal Gland ,Pineal gland ,0302 clinical medicine ,Arylalkylamine N-acetyltransferase ,Clock genes ,Melatonin receptors ,Reactive oxygen species ,Senescence ,Molecular Medicine ,Molecular Biology ,Melatonin ,Cholestasis ,Chemistry ,Biliary hyperplasia ,Pinealectomy ,medicine.anatomical_structure ,Liver ,030211 gastroenterology & hepatology ,Signal Transduction ,medicine.drug ,medicine.medical_specialty ,Kupffer Cells ,Primary Cell Culture ,digestive system ,Article ,Cholangiocyte ,Cell Line ,03 medical and health sciences ,arylalkylamine N-acetyltransferase ,clock genes ,melatonin receptors ,reactive oxygen species ,senescence ,Internal medicine ,Hepatic Stellate Cells ,medicine ,Animals ,Humans ,Arylalkylamine N-acetyltransferase, Clock genes, Melatonin receptors, Reactive oxygen species, Senescence ,Autocrine signalling ,Cell Proliferation ,Epithelial Cells ,Rats, Inbred F344 ,digestive system diseases ,Rats ,MicroRNAs ,Bile Ducts, Intrahepatic ,Endocrinology ,Gene Expression Regulation ,Hepatic stellate cell ,030217 neurology & neurosurgery - Abstract
Melatonin, a neuroendocrine hormone synthesized by the pineal gland and cholangiocytes, decreases biliary hyperplasia and liver fibrosis during cholestasis-induced biliary injury via melatonin-dependent autocrine signaling through increased biliary arylalkylamine N-acetyltransferase (AANAT) expression and melatonin secretion, downregulation of miR-200b and specific circadian clock genes. Melatonin synthesis is decreased by pinealectomy (PINX) or chronic exposure to light. We evaluated the effect of PINX or prolonged light exposure on melatonin-dependent modulation of biliary damage/ductular reaction/liver fibrosis. Studies were performed in male rats with/without BDL for 1 week with 12:12 h dark/light cycles, continuous light or after 1 week of PINX. The expression of AANAT and melatonin levels in serum and cholangiocyte supernatant were increased in BDL rats, while decreased in BDL rats following PINX or continuous light exposure. BDL-induced increase in serum chemistry, ductular reaction, liver fibrosis, inflammation, angiogenesis and ROS generation were significantly enhanced by PINX or light exposure. Concomitant with enhanced liver fibrosis, we observed increased biliary senescence and enhanced clock genes and miR-200b expression in total liver and cholangiocytes. In vitro, the expression of AANAT, clock genes and miR-200b was increased in PSC human cholangiocyte cell lines (hPSCL). The proliferation and activation of HHStecs (human hepatic stellate cell lines) were increased after stimulating with BDL cholangiocyte supernatant and further enhanced when stimulated with BDL rats following PINX or continuous light exposure cholangiocyte supernatant via intracellular ROS generation. Conclusion: Melatonin plays an important role in the protection of liver against cholestasis-induced damage and ductular reaction.
- Published
- 2019
24. miR-34a-dependent overexpression of Per1 decreases cholangiocarcinoma growth
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Ying Wan, Julie Venter, Heather Francis, John Greene, Yuyan Han, Cynthia J. Meininger, Laurent Ehrlich, Jerome P. Trzeciakowski, Holly Standeford, Gianfranco Alpini, Shannon Glaser, Nan Wu, and Fanyin Meng
- Subjects
Male ,0301 basic medicine ,endocrine system ,Programmed cell death ,Angiogenesis ,CLOCK Proteins ,Biology ,Article ,Cholangiocarcinoma ,Mice ,03 medical and health sciences ,Cell Line, Tumor ,microRNA ,Animals ,Humans ,Neoplasm Invasiveness ,Cell Proliferation ,Mice, Inbred BALB C ,Cyclin-dependent kinase 1 ,Hepatology ,Cell growth ,Period Circadian Proteins ,Cell cycle ,Circadian Rhythm ,MicroRNAs ,030104 developmental biology ,Bile Duct Neoplasms ,Cancer research ,PER1 - Abstract
Background & Aims Disruption of circadian rhythm is associated with cancer development and progression. MicroRNAs (miRNAs) are a class of small non-coding RNAs that trigger mRNA translation inhibition. We aimed to evaluate the role of Per1 and related miRNAs in cholangiocarcinoma growth. Methods The expression of clock genes was evaluated in human cholangiocarcinoma tissue arrays and cholangiocarcinoma lines. The rhythmic expression of clock genes was evaluated in cholangiocarcinoma cells and H69 (non-malignant cholangiocytes) by qPCR. We measured cell proliferation, cell cycle and apoptosis in Mz-ChA-1 cells after Per1 overexpression. We examined tumor growth in vivo after injection of Per1 overexpressing cells. We verified miRNAs that targets Per1 . The circadian rhythm of miR-34a was evaluated in cholangiocarcinoma and H69 cells. We evaluated cell proliferation, apoptosis and invasion after inhibition of miR-34a in vitro , and the potential molecular mechanisms by mRNA profiling after overexpression of Per1 . Results Expression of Per1 was decreased in cholangiocarcinoma. The circadian rhythm of Per1 expression was lost in cholangiocarcinoma cells. Decreased cell proliferation, lower G2/M arrest, and enhanced apoptosis were shown in Per1 overexpressing cells. An in vivo study revealed decreased tumor growth, decreased proliferation, angiogenesis and metastasis after overexpressing Per1 . Per1 was verified as a target of miR-34a. miR-34a was rhythmically expressed in cholangiocarcinoma cells and H69. The inhibition of miR-34a decreased proliferation, migration and invasion in cholangiocarcinoma cells. mRNA profiling has shown that overexpression of Per1 inhibits cell growth through regulation of multiple cancer-related pathways, such as cell cycle, cell growth and apoptosis pathways. Conclusions Disruption of circadian rhythms of clock genes contribute to the malignant phenotypes of human cholangiocarcinoma. Lay Summary The current study is about how biological clock and its regulators affect the bile duct tumor growth. The disruption of biological clock has a negative impact in different cancers. Per1 is a gene that is involved in maintaining the biological clock and show 24h oscillation. Reduced levels of Per1 and disruption of 24h circadian rhythm was found in bile duct cancer cells. Therefore, a genetic modified bile duct cancer cells was created. It has a higher level of Per1 expression and partially recovered circadian rhythm. Those genetic modified cells also displayed slower cell growth or higher rate of cell death. We also used mice model that lack of immune system to show that our genetic modified bile duct cells form smaller tumor. In addition, we tried to see how Per1 is communicating with other genes in regarding of controlling the tumor growth. We found Per1 is regulated by microRNA-34a, a small non-coding RNA that directly binds to genes and inhibit gene expression. Decreased level of miR-34a has also significantly reduced tumor growth through controlling the cell growth and cell death balance. Therefore bile duct cancer patients may be treated with miR-34a inhibitor or Per1 stimulator in the future.
- Published
- 2016
25. Ischemia reperfusion of the hepatic artery induces the functional damage of large bile ducts by changes in the expression of angiogenic factors
- Author
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Antonella Vetuschi, Shannon Glaser, Heather Francis, Romina Mancinelli, Paolo Onori, Eugenio Gaudio, Gianfranco Alpini, Antonio Franchitto, Julie Venter, Roberta Sferra, Luigi Pannarale, Fanyin Meng, and Guido Carpino
- Subjects
Male ,Vascular Endothelial Growth Factor A ,Pathology ,Time Factors ,Physiology ,medicine.medical_treatment ,Liver and Biliary Tract Physiology/Pathophysiology ,Vascular Endothelial Growth Factor C ,Apoptosis ,Liver transplantation ,Gastroenterology ,Hepatic Artery ,Cyclic AMP ,Angiogenic Proteins ,Cells, Cultured ,Cholestasis ,Cell Hypoxia ,Up-Regulation ,Reperfusion Injury ,Liver Circulation ,Signal Transduction ,medicine.medical_specialty ,angiogenic factors ,Ischemia ,Biology ,liver ,Cholangiocyte ,Paracrine signalling ,Secretin ,Physiology (medical) ,Internal medicine ,medicine ,Animals ,biliary tree ,RNA, Messenger ,cholangiocytes ,Cell Proliferation ,transplantation ,Hepatology ,Vascular Endothelial Growth Factor Receptor-3 ,medicine.disease ,Vascular Endothelial Growth Factor Receptor-2 ,Rats, Inbred F344 ,Transplantation ,Disease Models, Animal ,Bile Ducts, Intrahepatic ,Reperfusion injury - Abstract
Liver transplantation and cholangiocarcinoma induce biliary dysfunction following ischemia reperfusion (IR). The function of the intrahepatic biliary tree is regulated by both autocrine and paracrine factors. The aim of the study was to demonstrate that IR-induced damage of cholangiocytes is associated with altered expression of biliary angiogenic factors. Normal and bile duct ligation rats underwent 24-h sham or hepatic reperfusion after 30 min of transient occlusion of the hepatic artery (HAIR) or portal vein (PVIR) before collecting liver blocks and cholangiocyte RNA or protein. We evaluated liver histology, biliary apoptosis, proliferation and expression of VEGF-A/C, VEGFR-2/3, Ang-1/2, and Tie-1/2 in liver sections and isolated small and large cholangiocytes. Normal rat intrahepatic cholangiocyte cultures (NRICC) were maintained under standard conditions in normoxic or under a hypoxic atmosphere for 4 h and then transferred to normal conditions for selected times. Subsequently, we measured changes in biliary proliferation and apoptosis and the expression of VEGF-A/C and VEGFR-2/3. In vivo, HAIR (but not PVIR) induced damage of large bile ducts and decreased proliferation and secretin-stimulated cAMP levels. HAIR-induced damage of large bile ducts was associated with increased expression of VEGF-A/C, VEGFR-2/3, Ang-1/2, and Tie-1/2. In vitro, under hypoxic conditions, there was increased apoptosis and reduced proliferation of NRICC concomitant with enhanced expression of VEGF-A/C and VEGFR-2/3. The functional damage of large bile ducts by HAIR and hypoxia is associated with increased expression of angiogenic factors in small cholangiocytes, presumably due to a compensatory mechanism in response to biliary damage.
- Published
- 2015
26. Correction to: Knockout of α-calcitonin gene-related peptide attenuates cholestatic liver injury by differentially regulating cellular senescence of hepatic stellate cells and cholangiocytes
- Author
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Nan Wu, Q Huang, G Alpini, L Ceci, Ying Wan, Shannon Glaser, K Kyritsi, H. Francis, Pietro Invernizzi, L Chen, F Bernuzzi, F Meng, A Sybenga, T Zhou, P Baker, Julie Venter, and C Wu
- Subjects
Liver injury ,Chemistry ,medicine ,Hepatic stellate cell ,Cellular senescence ,Cell Biology ,Calcitonin gene-related peptide ,medicine.disease ,Molecular Biology ,Pathology and Forensic Medicine ,Cell biology - Published
- 2020
27. Opposite effects of knocking out MT1 and MT2 melatonin receptor on senescence and fibrosis of cholangiocytes and hepatic stellate cells during cholestatic liver injury
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Paolo Onori, Shannon Glaser, Tianhao Zhou, Julie Venter, Antonio Franchitto, Francesca Bernuzzi, Konstantina Kyritsi, Gianfranco Alpini, Eugenio Gaudio, Keisaku Sato, Heather Francis, Fanyin Meng, Nan Wu, and Pietro Invernizzi
- Subjects
Liver injury ,Senescence ,medicine.medical_specialty ,business.industry ,medicine.disease ,Biochemistry ,Melatonin receptor ,Endocrinology ,Fibrosis ,Internal medicine ,Genetics ,medicine ,Hepatic stellate cell ,business ,Molecular Biology ,Biotechnology - Published
- 2018
28. Melatonin or Dark Therapy Reduce Biliary Damage, Inflammation and Liver Fibrosis in a Murine Model of Early Stage Primary Biliary Cholangitis
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Julie Venter, Gianfranco Alpini, Heather Francis, Lindsey Kennedy, Nan Wu, Konstantina Kyritsi, Fanyin Meng, Shannon Glaser, Eugenio Gaudio, and Tianhao Zhou
- Subjects
Pathology ,medicine.medical_specialty ,business.industry ,Liver fibrosis ,Inflammation ,Biochemistry ,Melatonin ,Dark therapy ,Murine model ,Genetics ,medicine ,medicine.symptom ,Stage (cooking) ,business ,Molecular Biology ,Biotechnology ,medicine.drug - Published
- 2018
29. Knockout of secretin receptor reduces biliary damage and liver fibrosis in Mdr2
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Tianhao, Zhou, Nan, Wu, Fanyin, Meng, Julie, Venter, Thao K, Giang, Heather, Francis, Konstantina, Kyritsi, Chaodong, Wu, Antonio, Franchitto, Domenico, Alvaro, Marco, Marzioni, Paolo, Onori, Romina, Mancinelli, Eugenio, Gaudio, Shannon, Glaser, and Gianfranco, Alpini
- Subjects
Liver Cirrhosis ,Male ,Mice, Knockout ,ATP Binding Cassette Transporter, Subfamily B ,Cholangitis, Sclerosing ,Neovascularization, Physiologic ,Article ,Receptors, G-Protein-Coupled ,Receptors, Gastrointestinal Hormone ,Transforming Growth Factor beta1 ,Disease Models, Animal ,MicroRNAs ,Liver ,Secretin ,Paracrine Communication ,Hepatic Stellate Cells ,Animals ,Humans ,Cellular Senescence - Abstract
Secretin receptor (SR), only expressed by cholangiocytes, plays a key role in the regulation of biliary damage and liver fibrosis. The aim of this study was to determine the effects of genetic depletion of SR in Mdr2−/− mice on intrahepatic biliary mass, liver fibrosis, senescence and angiogenesis. 12 wk SR−/−, Mdr2−/− and SR−/−/Mdr2−/− mice with corresponding wild-type mice were used for the in vivo studies. Immunohistochemistry or immunofluorescence was performed in liver sections for (i) biliary expression of SR; (ii) hematoxylin and eosin; (iii) intrahepatic biliary mass by CK-19; (iv) fibrosis by Col1a1 and α-SMA; (v) senescence by SA-β-gal and p16; and (vi) angiogenesis by VEGF-A and CD31. Secretin (Sct) and TGF-β1 levels were measured in serum and cholangiocyte supernatant by ELISA. In total liver, isolated cholangiocytes or HSCs, we evaluated the expression of fibrosis markers (FN-1 and Col1a1); senescence markers (p16 and CCL2); microRNA 125b and angiogenesis markers (VEGF-A, VEGFR-2, CD31 and vWF) by immunoblots and/or qPCR. In vitro, we measured the paracrine effect of cholangiocyte supernatant on the expression of senescent and fibrosis markers in human hepatic stellate cells (HHSteCs). The increased level of ductular reaction, fibrosis and angiogenesis in Mdr2−/− mice was reduced in SR−/−/Mdr2−/− mice. Enhanced senescence levels in cholangiocytes from Mdr2−/− mice were reversed to normal in SR−/−/Mdr2−/− mice. However, senescence was decreased in HSCs from Mdr2−/− mice but returned to normal values in SR−/−/Mdr2−/− mice. In vitro treatment of HHSteCs with supernatant from cholangiocyte lacking SR (containing lower biliary levels of Sct-dependent TGF-β1) have decreased fibrotic reaction and increased cellular senescence. Sct-induced TGF-β1 secretion was mediated by microRNA 125b. Our data suggest that differential modulation of angiogenesis-dependent senescence of cholangiocytes and HSCs may be important for the treatment of liver fibrosis in cholangiopathies.
- Published
- 2018
30. The secretin/secretin receptor axis modulates ductular reaction and liver fibrosis through changes in transforming growth factor (TGF)-β1-mediated biliary senescence
- Author
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Konstantina Kyritsi, Nan Wu, Thao Giang, Tianhao Zhou, Gianfranco Alpini, Romina Mancinelli, Paolo Onori, Shannon Glaser, Fanyin Meng, Domenico Alvaro, Heather Francis, Julie Venter, Chaodong Wu, Antonio Franchitto, and Eugenio Gaudio
- Subjects
Liver Cirrhosis ,Male ,0301 basic medicine ,Senescence ,medicine.medical_specialty ,Kupffer Cells ,digestive system ,Article ,Cholangiocyte ,Receptors, G-Protein-Coupled ,Receptors, Gastrointestinal Hormone ,Pathology and Forensic Medicine ,Secretin ,Transforming Growth Factor beta1 ,secretin/secretin receptor axis ,TGFβ1 ,biliary senescence ,liver fibrosis ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,medicine ,Animals ,RNA, Messenger ,Cellular Senescence ,Mice, Knockout ,Liver injury ,Chemistry ,Organ Size ,medicine.disease ,digestive system diseases ,Mice, Inbred C57BL ,030104 developmental biology ,Endocrinology ,Hepatic stellate cell ,Secretin receptor ,030211 gastroenterology & hepatology ,Bile Ducts ,Hepatic fibrosis ,Transforming growth factor - Abstract
Activation of the secretin (Sct)/secretin receptor (SR) axis stimulates ductular reaction and liver fibrosis, which are hallmarks of cholangiopathies. Our aim was to define the role of Sct-regulated cellular senescence, and we demonstrated that both ductular reaction and liver fibrosis are significantly reduced in Sct(−/−), SR(−/−), and Sct(−/−)/SR(−/−) bile duct ligated (BDL) mice compared with BDL wild-type mice. The reduction in hepatic fibrosis in Sct(−/−), SR(−/−), and Sct(−/−)/SR(−/−) BDL mice was accompanied by reduced transforming growth factor-β1 levels in serum and cholangiocyte supernatant, as well as decreased expression of markers of cellular senescence in cholangiocytes in contrast to enhanced cellular senescence in hepatic stellate cells compared with BDL wild-type mice. Secretin directly stimulated the senescence of cholangiocytes and regulated, by a paracrine mechanism, the senescence of hepatic stellate cells and liver fibrosis via modulation of transforming growth factor-β1 biliary secretion. Targeting senescent cholangiocytes may represent a novel therapeutic approach for ameliorating hepatic fibrosis during cholestatic liver injury.
- Published
- 2018
31. Knockout of secretin receptor reduces biliary damage and liver fibrosis in Mdr2−/−mice by diminishing senescence of cholangiocytes
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Julie Venter, Nan Wu, Heather Francis, Marco Marzioni, Domenico Alvaro, Gianfranco Alpini, Konstantina Kyritsi, Thao Giang, Shannon Glaser, Romina Mancinelli, Paolo Onori, Fanyin Meng, Chaodong Wu, Antonio Franchitto, Tianhao Zhou, and Eugenio Gaudio
- Subjects
0301 basic medicine ,Senescence ,molecular Biology ,cell Biology ,Chemistry ,Angiogenesis ,Cell Biology ,medicine.disease ,Molecular biology ,Cholangiocyte ,3. Good health ,Pathology and Forensic Medicine ,Secretin ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Fibrosis ,medicine ,Hepatic stellate cell ,Secretin receptor ,030211 gastroenterology & hepatology ,Receptor ,Molecular Biology - Abstract
Secretin receptor (SR), only expressed by cholangiocytes, plays a key role in the regulation of biliary damage and liver fibrosis. The aim of this study was to determine the effects of genetic depletion of SR in Mdr2−/− mice on intrahepatic biliary mass, liver fibrosis, senescence, and angiogenesis. 12 wk SR−/−, Mdr2−/−, and SR−/−/Mdr2−/− mice with corresponding wild-type mice were used for the in vivo studies. Immunohistochemistry or immunofluorescence was performed in liver sections for (i) biliary expression of SR; (ii) hematoxylin and eosin; (iii) intrahepatic biliary mass by CK-19; (iv) fibrosis by Col1a1 and α-SMA; (v) senescence by SA-β-gal and p16; and (vi) angiogenesis by VEGF-A and CD31. Secretin (Sct) and TGF-β1 levels were measured in serum and cholangiocyte supernatant by ELISA. In total liver, isolated cholangiocytes or HSCs, we evaluated the expression of fibrosis markers (FN-1 and Col1a1); senescence markers (p16 and CCL2); microRNA 125b and angiogenesis markers (VEGF-A, VEGFR-2, CD31, and vWF) by immunoblots and/or qPCR. In vitro, we measured the paracrine effect of cholangiocyte supernatant on the expression of senescent and fibrosis markers in human hepatic stellate cells (HHSteCs). The increased level of ductular reaction, fibrosis, and angiogenesis in Mdr2−/− mice was reduced in SR−/−/Mdr2−/− mice. Enhanced senescence levels in cholangiocytes from Mdr2−/− mice were reversed to normal in SR−/−/Mdr2−/− mice. However, senescence was decreased in HSCs from Mdr2−/− mice but returned to normal values in SR−/−/Mdr2−/− mice. In vitro treatment of HHSteCs with supernatant from cholangiocyte lacking SR (containing lower biliary levels of Sct-dependent TGF-β1) have decreased fibrotic reaction and increased cellular senescence. Sct-induced TGF-β1 secretion was mediated by microRNA 125b. Our data suggest that differential modulation of angiogenesis-dependent senescence of cholangiocytes and HSCs may be important for the treatment of liver fibrosis in cholangiopathies.
- Published
- 2018
32. Gonadotropin-Releasing Hormone Stimulates Biliary Proliferation by Paracrine/Autocrine Mechanisms
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Yuyan Han, Paolo Onori, Eugenio Gaudio, Fanyin Meng, Heather Francis, Lindsey Kennedy, Debolina Ray, Antonio Franchitto, Julie Venter, Sharon DeMorrow, Shannon Glaser, Romina Mancinelli, Matthew McMillin, and Gianfranco Alpini
- Subjects
Liver Cirrhosis ,Male ,medicine.medical_specialty ,endocrine system ,Hypothalamus ,Cholangiocyte proliferation ,Fluorescent Antibody Technique ,Intrahepatic bile ducts ,Inositol 1,4,5-Trisphosphate ,Gonadotropin-releasing hormone ,Biology ,Autocrine Communication ,liver ,Cholangiocyte ,Cell Line ,Morpholinos ,Pathology and Forensic Medicine ,Mice ,Paracrine signalling ,Internal medicine ,Paracrine Communication ,Cyclic AMP ,medicine ,Animals ,biliary tree ,gonadotropin-releasing hormone ,Gene Silencing ,Autocrine signalling ,Cell Proliferation ,GNRHR ,Regular Article ,Rats, Inbred F344 ,Bile Ducts, Intrahepatic ,Endocrinology ,Receptors, LHRH ,hormones, hormone substitutes, and hormone antagonists - Abstract
During cholestatic liver disease, there is dysregulation in the balance between biliary growth and loss in bile duct–ligated (BDL) rats modulated by neuroendocrine peptides via autocrine/paracrine pathways. Gonadotropin-releasing hormone (GnRH) is a trophic peptide hormone that modulates reproductive function and proliferation in many cell types. We evaluated the autocrine role of GnRH in the regulation of cholangiocyte proliferation. The expression of GnRH receptors was assessed in a normal mouse cholangiocyte cell line (NMC), sham, and BDL rats. The effect of GnRH administration was evaluated in normal rats and in NMC. GnRH-induced biliary proliferation was evaluated by changes in intrahepatic bile duct mass and the expression of proliferation and function markers. The expression and secretion of GnRH in NMC and isolated cholangiocytes was assessed. GnRH receptor subtypes GnRHR 1 and GnRHR 2 were expressed in cholangiocytes. Treatment with GnRH increased intrahepatic bile duct mass as well as proliferation and function markers in cholangiocytes. Transient knockdown and pharmacologic inhibition of GnRHR 1 in NMC decreased proliferation. BDL cholangiocytes had increased expression of GnRH compared with normal rats, accompanied by increased GnRH secretion. In vivo and in vitro knockdown of GnRH decreased intrahepatic bile duct mass/cholangiocyte proliferation and fibrosis. GnRH secreted by cholangiocytes promotes biliary proliferation via an autocrine pathway. Disruption of GnRH/GnRHR signaling may be important for the management of cholestatic liver diseases.
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- 2015
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33. Regulation of the Extrinsic Apoptotic Pathway by MicroRNA-21 in Alcoholic Liver Injury
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Phillip Levine, Fanyin Meng, Shannon Glaser, Li Huang, Tianhao Zhou, Heather Francis, Gianfranco Alpini, Yuyan Han, Fuquan Yang, Jennifer McCarra, Lindsey Kennedy, Xiuping Liu, Jia Ming Lai, Julie Venter, Kelly McDaniel, and Chang-Gong Liu
- Subjects
STAT3 Transcription Factor ,endocrine system ,congenital, hereditary, and neonatal diseases and abnormalities ,Alcoholic liver disease ,endocrine system diseases ,Apoptosis ,Biochemistry ,Fas ligand ,Mice ,microRNA ,Hepatic Stellate Cells ,medicine ,Animals ,Humans ,STAT3 ,Liver Diseases, Alcoholic ,Molecular Biology ,Regulator gene ,Liver injury ,biology ,Interleukin-6 ,nutritional and metabolic diseases ,Cell Biology ,medicine.disease ,Molecular biology ,Up-Regulation ,Mice, Inbred C57BL ,MicroRNAs ,Real-time polymerase chain reaction ,Liver ,Cancer research ,Hepatic stellate cell ,biology.protein ,Signal Transduction - Abstract
IL-6/Stat3 is associated with the regulation of transcription of key cellular regulatory genes (microRNAs) during different types of liver injury. This study evaluated the role of IL-6/Stat3 in regulating miRNA and miR-21 in alcoholic liver disease. By microarray, we identified that ethanol feeding significantly up-regulated 0.8% of known microRNAs in mouse liver compared with controls, including miR-21. Similarly, the treatment of normal human hepatocytes (N-Heps) and hepatic stellate cells (HSCs) with ethanol and IL-6 significantly increased miR-21 expression. Overexpression of miR-21 decreased ethanol-induced apoptosis in both N-Heps and HSCs. The expression level of miR-21 was significantly increased after Stat3 activation in N-Heps and HSCs, in support of the concept that the 5'-promoter region of miR-21 is regulated by Stat3. Using real time PCR, we confirmed that miR-21 activation is associated with ethanol-linked Stat3 binding of the miR-21 promoter. A combination of bioinformatics, PCR array, dual-luciferase reporter assay, and Western blot analysis revealed that Fas ligand (TNF superfamily, member 6) (FASLG) and death receptor 5 (DR5) are the direct targets of miR-21. Furthermore, inhibition of miR-21 by specific Vivo-Morpholino and knock-out of IL-6 in ethanol-treated mice also increased the expression of DR5 and FASLG in vivo during alcoholic liver injury. The identification of miR-21 as an important regulator of hepatic cell survival, transformation, and remodeling in vitro, as well as its upstream modulators and downstream targets, will provide insight into the involvement of altered miRNA expression in contributing to alcoholic liver disease progression and testing novel therapeutic approaches for human alcoholic liver diseases.
- Published
- 2014
34. The role of the secretin/secretin receptor axis in inflammatory cholangiocyte communication via extracellular vesicles
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Julie Venter, Thao Giang, Gianfranco Alpini, Fanyin Meng, Keisaku Sato, and Shannon Glaser
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0301 basic medicine ,medicine.medical_specialty ,Multidisciplinary ,Chemistry ,lcsh:R ,Secretin receptor family ,lcsh:Medicine ,Secretin family ,Cholangiocyte ,Article ,Proinflammatory cytokine ,Cell biology ,Secretin ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Endocrinology ,Internal medicine ,medicine ,Secretin receptor ,030211 gastroenterology & hepatology ,Secretion ,Hormone metabolism ,lcsh:Q ,lcsh:Science - Abstract
Small and large intrahepatic bile ducts consist of small and large cholangiocytes, respectively, and these cholangiocytes have different morphology and functions. The gastrointestinal peptide hormone, secretin (SCT) that binds to secretin receptor (SR), is a key mediator in cholangiocyte pathophysiology. Extracellular vesicles (EVs) are membrane-bound vesicles and cell-cell EV communication is recognized as an important factor in liver pathology, although EV communication between cholangiocytes is not identified to date. Cholangiocytes secrete proinflammatory cytokines during bacterial infection leading to biliary inflammation and hyperplasia. We demonstrate that cholangiocytes stimulated with lipopolysaccharide (LPS), which is a membrane component of gram-negative bacteria, secrete more EVs than cholangiocytes incubated with vehicle. These LPS-derived EVs induce inflammatory responses in other cholangiocytes including elevated cytokine production and cell proliferation. Large but not small cholangiocytes show inflammatory responses against large but not small cholangiocyte-derived EVs. Large cholangiocytes with knocked down either SCT or SR by short hairpin RNAs show reduced EV secretion during LPS stimulation, and EVs isolated from SCT or SR knocked down cholangiocytes fail to induce inflammatory reactions in control large cholangiocytes. This study identifies cholangiocyte EV communication during LPS stimulation, and demonstrates that the SCT/SR axis may be important for this event.
- Published
- 2017
35. The Hippo signaling functions through the Notch signaling to regulate intrahepatic bile duct development in mammals
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Gianfranco Alpini, Duojia Pan, Ying Wan, Tiaohao Zhou, Quy Nguyen, Haibo Bai, Sharon DeMorrow, Nan Wu, Julie Venter, Gabriel Frampton, Fanyin Meng, and Shannon Glaser
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0301 basic medicine ,Male ,medicine.medical_specialty ,endocrine system ,Neurofibromatosis 2 ,Cell ,Notch signaling pathway ,Intrahepatic bile ducts ,Biology ,Protein Serine-Threonine Kinases ,Article ,Pathology and Forensic Medicine ,03 medical and health sciences ,Mice ,Mediator ,Internal medicine ,medicine ,Animals ,Hippo Signaling Pathway ,Receptor, Notch2 ,Molecular Biology ,Mice, Knockout ,Hippo signaling pathway ,Bile duct ,Effector ,Cell Biology ,Cell biology ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,Bile Ducts, Intrahepatic ,Hippo signaling ,Female - Abstract
The Hippo signaling pathway and the Notch signaling pathway are evolutionary conserved signaling cascades that have important roles in embryonic development of many organs. In murine liver, disruption of either pathway impairs intrahepatic bile duct development. Recent studies suggested that the Notch signaling receptor Notch2 is a direct transcriptional target of the Hippo signaling pathway effector YAP, and the Notch signaling is a major mediator of the Hippo signaling in maintaining biliary cell characteristics in adult mice. However, it remains to be determined whether the Hippo signaling pathway functions through the Notch signaling in intrahepatic bile duct development. We found that loss of the Hippo signaling pathway tumor suppressor Nf2 resulted in increased expression levels of the Notch signaling pathway receptor Notch2 in cholangiocytes but not in hepatocytes. When knocking down Notch2 on the background of Nf2 deficiency in mouse livers, the excessive bile duct development induced by Nf2 deficiency was suppressed by heterozygous and homozygous deletion of Notch2 in a dose-dependent manner. These results implicated that Notch signaling is one of the downstream effectors of the Hippo signaling pathway in regulating intrahepatic bile duct development.
- Published
- 2017
36. Forkhead box A2 regulates biliary heterogeneity and senescence during cholestatic liver injury in mice‡
- Author
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Julie Venter, Heather Francis, Keisaku Sato, Ying Wan, Paolo Onori, Gianfranco Alpini, Qiaobing Huang, Tianhao Zhou, Nan Wu, Konstantina Kyritsi, Pietro Invernizzi, Fanyin Meng, Francesca Bernuzzi, Kelly McDaniel, Shannon Glaser, Eugenio Gaudio, Mcdaniel, K, Meng, F, Wu, N, Sato, K, Venter, J, Bernuzzi, F, Invernizzi, P, Zhou, T, Kyritsi, K, Wan, Y, Huang, Q, Onori, P, Francis, H, Gaudio, E, Glaser, S, and Alpini, G
- Subjects
0301 basic medicine ,Pathology ,Aging ,biliary heterogeneity ,Cellular differentiation ,Cell Communication ,Mice ,0302 clinical medicine ,Fibrosis ,cellular senescence ,Liver injury ,Aged, 80 and over ,Mice, Knockout ,DNA methylation ,Cholestasis ,Middle Aged ,Liver ,Knockout mouse ,primary sclerosing cholangiti ,Hepatocyte Nuclear Factor 3-beta ,030211 gastroenterology & hepatology ,Senescence ,Adult ,medicine.medical_specialty ,Adolescent ,Down-Regulation ,hepatology ,cholestatic liver injury ,foxA2 ,Biology ,Real-Time Polymerase Chain Reaction ,digestive system ,Article ,Sampling Studies ,Primary sclerosing cholangitis ,03 medical and health sciences ,Genetic Heterogeneity ,FoxA2 ,medicine ,Hepatic Stellate Cells ,Animals ,Humans ,Progenitor cell ,Aged ,Hepatology ,medicine.disease ,digestive system diseases ,Disease Models, Animal ,030104 developmental biology ,Gene Expression Regulation ,Hepatic stellate cell ,Hepatocytes ,Bile Ducts - Abstract
Biliary-committed progenitor cells (small mouse cholangiocytes; SMCCs) from small bile ducts are more resistant to hepatobiliary injury than large mouse cholangiocytes (LGCCs) from large bile ducts. The definitive endoderm marker, forkhead box A2 (FoxA2), is the key transcriptional factor that regulates cell differentiation and tissue regeneration. Our aim was to characterize the translational role of FoxA2 during cholestatic liver injury. Messenger RNA expression in SMCCs and LGCCs was assessed by polymerase chain reaction (PCR) array analysis. Liver tissues and hepatic stellate cells (HSCs) from primary sclerosing cholangitis (PSC) and primary biliary cholangitis (PBC) patients were tested by real-time PCR for methylation, senescence, and fibrosis markers. Bile duct ligation (BDL) and multidrug resistance protein 2 (MDR2) knockout mice (MDR2-/-) were used as animal models of cholestatic liver injury with or without healthy transplanted large or small cholangiocytes. We demonstrated that FoxA2 was notably enhanced in murine liver progenitor cells and SMCCs and was silenced in human PSC and PBC liver tissues relative to respective controls that are correlated with the epigenetic methylation enzymes, DNA methyltransferase (DNMT) 1 and DNMT3B. Serum alanine aminotransferase and aspartate aminotransferase levels in nonobese diabetic/severe combined immunodeficiency mice engrafted with SMCCs post-BDL showed significant changes compared to vehicle-treated mice, along with improved liver fibrosis. Enhanced expression of FoxA2 was observed in BDL mouse liver after SMCC cell therapy. Furthermore, activation of fibrosis signaling pathways were observed in BDL/MDR2-/- mouse liver as well as in isolated HSCs by laser capture microdissection, and these signals were recovered along with reduced hepatic senescence and enhanced hepatic stellate cellular senescence after SMCC engraft. Conclusion: The definitive endoderm marker and the positive regulator of biliary development, FoxA2, mediates the therapeutic effect of biliary-committed progenitor cells during cholestatic liver injury.
- Published
- 2017
37. Secretin receptor antagonist treatment reduces biliary damage and liver fibrosis in a mouse model of early stage primary biliary cholangitis (PBC), but not advanced PBC
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Lindsey, Kennedy, Francis, Heather L., Julie, Venter, Laura, Hargrove, Nan, Wu, Pietro, Invernizzi, Eric Gershwin, M., Francesca, Bernuzzi, Fanyin, Meng, Marco, Marzioni, Onori, Paolo, Franchitto, Antonio, Alvaro, Domenico, Gaudio, Eugenio, Glaser, Shannon S., and Gianfranco, Alpini
- Subjects
primary biliary cholangitis ,biliary tree ,liver - Published
- 2017
38. Differential impact of MT1 and MT2 melatonin receptor deletion on biliary proliferation, senescence, and liver fibrosis during cholestatic liver injury
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Nan, Wu, Tina, Kyritsi, Pietro, Invernizzi, Julie, Venter, Francesca, Bernuzzi, Fanyin, Meng, Keisaku, Sato, Gaudio, Eugenio, Francis, Heather L., Tianhao, Zhou, Onori, Paolo, Franchitto, Antonio, Gianfranco, Alpini, and Glaser, Shannon S.
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liver ,biliary tree ,melatonin - Published
- 2017
39. Overexpression of membrane metalloendopeptidase inhibits substance P stimulation of cholangiocarcinoma growth
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Kelly McDaniel, Syeda H. Afroze, Lindsey Kennedy, Shannon Glaser, Sharon DeMorrow, Micheleine Guerrier, Yuyan Han, Gianfranco Alpini, Laura Hargrove, Shanika Avila, Heather Francis, Debolina Ray, Holly Standeford, Julie Venter, Gabriel Frampton, Fanyin Meng, Matthew McMillin, and Morgan Quezada
- Subjects
Male ,Vascular Endothelial Growth Factor A ,medicine.medical_specialty ,Time Factors ,Physiology ,Mice, Nude ,Substance P ,Biology ,Transfection ,Gene Expression Regulation, Enzymologic ,Cholangiocyte ,Cholangiocarcinoma ,Mice ,Neurokinin-1 Receptor Antagonists ,In vivo ,Cell Line, Tumor ,Proliferating Cell Nuclear Antigen ,Physiology (medical) ,Internal medicine ,parasitic diseases ,medicine ,Animals ,Humans ,Secretion ,Autocrine signalling ,Neprilysin ,Cell Proliferation ,Keratin-19 ,Mice, Inbred BALB C ,Dose-Response Relationship, Drug ,Hepatology ,Cell growth ,Gastroenterology ,Receptors, Neurokinin-1 ,Xenograft Model Antitumor Assays ,Molecular biology ,Tumor Burden ,Up-Regulation ,Gene Expression Regulation, Neoplastic ,Liver and Biliary Tract ,Vascular endothelial growth factor A ,Bile Ducts, Intrahepatic ,Endocrinology ,Bile Duct Neoplasms ,Cell culture - Abstract
Substance P (SP) promotes cholangiocyte growth during cholestasis by activating its receptor, NK1R. SP is a proteolytic product of tachykinin (Tac1) and is deactivated by membrane metalloendopeptidase (MME). This study aimed to evaluate the functional role of SP in the regulation of cholangiocarcinoma (CCA) growth. NK1R, Tac1, and MME expression and SP secretion were assessed in human CCA cells and nonmalignant cholangiocytes. The proliferative effects of SP (in the absence/presence of the NK1R inhibitor, L-733,060) and of L-733,060 were evaluated. In vivo, the effect of L-733,060 treatment or MME overexpression on tumor growth was evaluated by using a xenograft model of CCA in nu/nu nude mice. The expression of Tac1, MME, NK1R, PCNA, CK-19, and VEGF-A was analyzed in the resulting tumors. Human CCA cell lines had increased expression of Tac1 and NK1R, along with reduced levels of MME compared with nonmalignant cholangiocytes, resulting in a subsequent increase in SP secretion. SP treatment increased CCA cell proliferation in vitro, which was blocked by L-733,060. Treatment with L-733,060 alone inhibited CCA proliferation in vitro and in vivo. Xenograft tumors derived from MME-overexpressed human Mz-ChA-1 CCA cells had a slower growth rate than those derived from control cells. Expression of PCNA, CK-19, and VEGF-A decreased, whereas MME expression increased in the xenograft tumors treated with L-733,060 or MME-overexpressed xenograft tumors compared with controls. The study suggests that SP secreted by CCA promotes CCA growth via autocrine pathway. Blockade of SP secretion and NK1R signaling may be important for the management of CCA.
- Published
- 2014
40. Melatonin inhibits hypothalamic gonadotropin-releasing hormone release and reduces biliary hyperplasia and fibrosis in cholestatic rats
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Konstantina Kyritsi, Thao Giang, Gianfranco Alpini, Nan Wu, Julie Venter, John F. Greene, Stephanie Grant, Matthew McMillin, Fanyin Meng, Tianhao Zhou, Shannon Glaser, Sharon DeMorrow, and Brandi Jefferson
- Subjects
0301 basic medicine ,Liver Cirrhosis ,medicine.medical_specialty ,endocrine system ,Physiology ,Gonadotropin-releasing hormone ,Biology ,Pineal Gland ,Cholangiocyte ,Melatonin ,Gonadotropin-Releasing Hormone ,03 medical and health sciences ,0302 clinical medicine ,Hormone Antagonists ,Cholestasis ,Fibrosis ,Physiology (medical) ,Internal medicine ,medicine ,Hepatic Stellate Cells ,Endocrine system ,Animals ,Humans ,Cell Proliferation ,Hyperplasia ,Hepatology ,Biliary hyperplasia ,Gastroenterology ,Central Nervous System Depressants ,medicine.disease ,Rats ,Disease Models, Animal ,030104 developmental biology ,Endocrinology ,Hepatic stellate cell ,030211 gastroenterology & hepatology ,Bile Ducts ,hormones, hormone substitutes, and hormone antagonists ,Receptors, LHRH ,medicine.drug ,Research Article - Abstract
Melatonin is a hormone produced by the pineal gland with increased circulating levels shown to inhibit biliary hyperplasia and fibrosis during cholestatic liver injury. Melatonin also has the capability to suppress the release of hypothalamic gonadotropin-releasing hormone (GnRH), a hormone that promotes cholangiocyte proliferation when serum levels are elevated. However, the interplay and contribution of neural melatonin and GnRH to cholangiocyte proliferation and fibrosis in bile duct-ligated (BDL) rats have not been investigated. To test this, cranial levels of melatonin were increased by implanting osmotic minipumps that performed an intracerebroventricular (ICV) infusion of melatonin or saline for 7 days starting at the time of BDL. Hypothalamic GnRH mRNA and cholangiocyte secretion of GnRH and melatonin were assessed. Cholangiocyte proliferation and fibrosis were measured. Primary human hepatic stellate cells (HSCs) were treated with cholangiocyte supernatants, GnRH, or the GnRH receptor antagonist cetrorelix acetate, and cell proliferation and fibrosis gene expression were assessed. Melatonin infusion reduced hypothalamic GnRH mRNA expression and led to decreased GnRH and increased melatonin secretion from cholangiocytes. Infusion of melatonin was found to reduce hepatic injury, cholangiocyte proliferation, and fibrosis during BDL-induced liver injury. HSCs supplemented with BDL cholangiocyte supernatant had increased proliferation, and this increase was reversed when HSCs were supplemented with supernatants from melatonin-infused rats. GnRH stimulated fibrosis gene expression in HSCs, and this was reversed by cetrorelix acetate cotreatment. Increasing bioavailability of melatonin in the brain may improve outcomes during cholestatic liver disease.NEW & NOTEWORTHY We have previously demonstrated that GnRH is expressed in cholangiocytes and promotes their proliferation during cholestasis. In addition, dark therapy, which increases melatonin, reduced cholangiocyte proliferation and fibrosis during cholestasis. This study expands these findings by investigating neural GnRH regulation by melatonin during BDL-induced cholestasis by infusing melatonin into the brain. Melatonin infusion reduced cholangiocyte proliferation and fibrosis, and these effects are due to GNRH receptor 1-dependent paracrine signaling between cholangiocytes and hepatic stellate cells.
- Published
- 2016
41. Sa1502 – Knockout (KO) of the Melatonin Receptor, Mt2, Enhances Alcohol-Induced Ductular Reaction (DR), Biliary Senescence and Hepatic Fibrosis During Alcoholic Liver Disease (ALD)
- Author
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Shannon Glaser, Lixian Chen, Nan Wu, Fanyin Meng, David C. Zawieja, Tianhao Zhou, Konstantina Kyritsi, Gianfranco Alpini, Tori White, Heather Francis, Bin Gao, Adrien Guillot, Ludovica Ceci, Julie Venter, Suthat Liangpunsakul, and Lindsey Kennedy
- Subjects
Senescence ,Alcoholic liver disease ,medicine.medical_specialty ,Hepatology ,business.industry ,Gastroenterology ,Alcohol ,medicine.disease ,Melatonin receptor ,chemistry.chemical_compound ,Endocrinology ,chemistry ,Internal medicine ,medicine ,Hepatic fibrosis ,business - Published
- 2019
42. Sa1510 – Increased Serotonin (5HT) Biliary Synthesis Due to Enhanced Expression of Tryptophan Hydroxylase1 (TPH1) and Reduced Monoamine-Oxidase-A (MAO-A) Expression is Couple with Alcohol-Induced Liver Injury (ALI)
- Author
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Tianhao Zhou, Suthat Liangpunsakul, Adrien Guillot, Nan Wu, Ludovica Ceci, Fanyin Meng, Shannon Glas, Lixian Chen, Tori White, Julie Venter, Heather Francis, Bin Gao, Gianfranco Alpini, Konstantina Kyritsi, and Lindsey Kennedy
- Subjects
Liver injury ,medicine.medical_specialty ,TPH1 ,Hepatology ,biology ,Gastroenterology ,Tryptophan ,Alcohol ,medicine.disease ,chemistry.chemical_compound ,Endocrinology ,chemistry ,Internal medicine ,medicine ,biology.protein ,Serotonin ,Monoamine oxidase A ,5-HT receptor - Published
- 2019
43. 772 – Extracellular Vesicles Isolated from Cholangiocytes Lacking the Secretin/Secretin Receptor Axis Attenuate Liver Fibrosis Via Cargo Micrornas in the Mdr2-/- Mouse Model of Primary Sclerosing Cholangitis
- Author
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Keisaku Sato, Heather Francis, Julie Venter, Gianfranco Alpini, Lindsey Kennedy, Fanyin Meng, and Shannon Glaser
- Subjects
Hepatology ,Chemistry ,Liver fibrosis ,microRNA ,Gastroenterology ,medicine ,Cancer research ,Secretin receptor ,medicine.disease ,Extracellular vesicles ,Primary sclerosing cholangitis ,Secretin - Published
- 2019
44. Mo1470 – Secretin/Secretin Receptor Signaling Modulates Biliary Immunobiology and Subsequent T Cell Migration in Early Stage Primary Biliary Cholangitis (PBC)
- Author
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Gianfranco Alpini, Julie Venter, Fanyin Meng, Lindsey Kennedy, David C. Zawieja, Heather Francis, Nan Wu, Pietro Invernizzi, Eric Gershwin, Shannon Glaser, and Luca Fabris
- Subjects
Hepatology ,business.industry ,Gastroenterology ,Cancer research ,T cell migration ,Medicine ,Secretin receptor ,Stage (cooking) ,business ,Secretin - Published
- 2019
45. Mo1428 – Role of the Aanat/Melatonin/Mt1/Mt2/Per-1 Axis in the Regulation of Biliary Damage and Liver Fibrosis in Cholestatic Mice
- Author
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Lindsey Kennedy, Paolo Onori, Heather Francis, Ludovica Ceci, Konstantina Kyritsi, Fanyin Meng, Nan Wu, Julie Venter, Eugenio Gaudio, Lixian Chen, Shannon Glaser, Gianfranco Alpini, Tianhao Zhou, and April O'Brien
- Subjects
Melatonin ,medicine.medical_specialty ,Endocrinology ,Hepatology ,business.industry ,AANAT ,Internal medicine ,Liver fibrosis ,Gastroenterology ,Medicine ,business ,medicine.drug - Published
- 2019
46. Mo1430 – The Proliferative Activity of Cholangiocytes is Regulated by Micrornas Carried in Extracellular Vesicles Secreted from Other Cholangiocytes
- Author
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Gianfranco Alpini, Fanyin Meng, Julie Venter, Lindsey Kennedy, Heather Francis, Shannon Glaser, and Keisaku Sato
- Subjects
Hepatology ,Chemistry ,microRNA ,Gastroenterology ,Extracellular vesicles ,Cell biology - Published
- 2019
47. Sa1520 – Knockdown of Stimulator of Interferon Genes (STING) Reduces Biliary Senescence and Liver Inflammation and Fibrosis in the Mdr2-/- Mouse Model of Primary Sclerosing Cholangitis (PSC)
- Author
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Tianhao Zhou, Julie Venter, Anatoliy A. Gashev, Heather Francis, Chaodong Wu, Lixian Chen, Konstantina Kyritsi, Lindsey Kennedy, David C. Zawieja, Gianfranco Alpini, Shannon Glaser, Ludovica Ceci, Nan Wu, and Fanyin Meng
- Subjects
Senescence ,Gene knockdown ,Hepatology ,business.industry ,Gastroenterology ,Inflammation ,medicine.disease ,Primary sclerosing cholangitis ,Sting ,Fibrosis ,Stimulator of interferon genes ,medicine ,Cancer research ,medicine.symptom ,business - Published
- 2019
48. Neuropeptide Y inhibits biliary hyperplasia of cholestatic rats by paracrine and autocrine mechanisms
- Author
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Gabriel Frampton, Fanyin Meng, Romina Mancinelli, Paolo Onori, Kelly McDaniel, Julie Venter, Dinorah Leyva-Illades, Gianfranco Alpini, Matthew A. Quinn, Shannon Glaser, Antonio Franchitto, Eugenio Gaudio, Hae Yong Pae, Heather Francis, and Sharon DeMorrow
- Subjects
Male ,medicine.medical_specialty ,Physiology ,proliferation ,Paracrine Communication ,Biology ,Autocrine Communication ,digestive system ,neurotransmitters ,Cell Line ,Paracrine signalling ,Proliferating Cell Nuclear Antigen ,Physiology (medical) ,Internal medicine ,mental disorders ,medicine ,Animals ,Homeostasis ,Neuropeptide Y ,RNA, Messenger ,Autocrine signalling ,Receptor ,Cell Proliferation ,Cholestasis ,Hyperplasia ,Hepatology ,biliary epithelium ,Biliary hyperplasia ,Gastroenterology ,Neuropeptide Y receptor ,Antibodies, Neutralizing ,Rats, Inbred F344 ,humanities ,Rats ,Receptors, Neuropeptide Y ,Liver and Biliary Tract ,Bile Ducts, Intrahepatic ,Endocrinology ,cell cycle ,Signal transduction ,Signal Transduction - Abstract
Neuropeptide Y (NPY) exerts its functions through six subtypes of receptors (Y1–Y6). Biliary homeostasis is regulated by several factors through autocrine/paracrine signaling. NPY inhibits cholangiocarcinoma growth; however, no information exists regarding the autocrine/paracrine role of NPY on biliary hyperplasia during cholestasis. The aims of this study were to determine: 1) the expression of NPY and Y1–Y5 in cholangiocytes and 2) the paracrine/autocrine effects of NPY on cholangiocyte proliferation. Normal or bile duct ligation (BDL) rats were treated with NPY, neutralizing anti-NPY antibody, or vehicle for 7 days. NPY and NPY receptor (NPYR) expression was assessed in liver sections and isolated cholangiocytes. NPY secretion was assessed in serum and bile from normal and BDL rats, as well as supernatants from normal and BDL cholangiocytes and normal rat cholangiocyte cell line [intrahepatic normal cholangiocyte culture (NRICC)]. We evaluated intrahepatic bile ductal mass (IBDM) in liver sections and proliferation in cholangiocytes. With the use of NRICC, the effects of NPY or anti-NPY antibody on cholangiocyte proliferation were determined. The expression of NPY and all NPYR were increased after BDL. NPY levels were lower in serum and cholangiocyte supernatant from BDL compared with normal rats. NPY secretion from NRICC was detected at both the basolateral and apical domains. Chronic NPY treatment decreased proliferating cellular nuclear antigen (PCNA) expression and IBDM in BDL rats. Administration of anti-NPY antibody to BDL rats increased cholangiocyte proliferation and IBDM. NPY treatment of NRICC decreased PCNA expression and increased the cell cycle arrest, whereas treatment with anti-NPY antibody increased proliferation. Therapies targeting NPY-mediated signaling may prove beneficial for the treatment of cholangiopathies.
- Published
- 2013
49. Recent advances in the morphological and functional heterogeneity of the biliary epithelium
- Author
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Paolo Onori, Shannon Glaser, Antonio Franchitto, Guido Carpino, Marco Marzioni, Domenico Alvaro, Julie Venter, Heather Francis, Yuyan Han, Gianfranco Alpini, Fanyin Meng, Eugenio Gaudio, and Kelly McDaniel
- Subjects
Biliary Tract Diseases ,Apoptosis ,camp ,Biology ,neurotransmitters ,Epithelium ,Article ,General Biochemistry, Genetics and Molecular Biology ,Gastrointestinal Hormones ,bile ducts ,gastrointestinal hormones ,vegf ,growth factors ,In vivo ,medicine ,Animals ,Humans ,Biliary epithelium ,Progenitor cell ,Cell Proliferation ,Cell growth ,Stem Cells ,Neuropeptides ,Phenotype ,Cell biology ,medicine.anatomical_structure ,Immunology ,Bile Ducts ,Stem cell ,Hormone - Abstract
This review focuses on the recent advances related to the heterogeneity of different-sized bile ducts with regard to the morphological and phenotypical characteristics, and the differential secretory, apoptotic and proliferative responses of small and large cholangiocytes to gastrointestinal hormones/peptides, neuropeptides and toxins. We describe several in vivo and in vitro models used for evaluating biliary heterogeneity. Subsequently, we discuss the heterogeneous proliferative and apoptotic responses of small and large cholangiocytes to liver injury and the mechanisms regulating the differentiation of small into large (more differentiated) cholangiocytes. Following a discussion on the heterogeneity of stem/progenitor cells in the biliary epithelium, we outline the heterogeneity of bile ducts in human cholangiopathies. After a summary section, we discuss the future perspectives that will further advance the field of the functional heterogeneity of the biliary epithelium.
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- 2013
50. Dysregulation of vitamin D3 synthesis leads to enhanced cholangiocarcinoma growth
- Author
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Evan Harnish, Julie Venter, Rachel Harris, Kimberly Baker, Laura Hargrove, Allyson Graf, Heather Francis, Kyle Hodges, Fanyin Meng, and Lindsey Kennedy
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Vitamin ,medicine.medical_specialty ,25-Hydroxyvitamin D3 1-alpha-hydroxylase ,Vitamin D3 24-Hydroxylase ,Calcitriol receptor ,Cholangiocarcinoma ,chemistry.chemical_compound ,CYP24A1 ,Cell Line, Tumor ,Internal medicine ,medicine ,Vitamin D and neurology ,Humans ,RNA, Messenger ,Receptor ,Cell Proliferation ,Cholecalciferol ,25-Hydroxyvitamin D3 1-alpha-Hydroxylase ,Hepatology ,business.industry ,Gastroenterology ,Vitamins ,digestive system diseases ,Protein Transport ,Bile Ducts, Intrahepatic ,Endocrinology ,Bile Duct Neoplasms ,chemistry ,Vitamin D3 Receptor ,Steroid Hydroxylases ,Receptors, Calcitriol ,business - Abstract
Background Cholangiocarcinoma is a deadly biliary tumour with limited treatment strategies. Vitamin (1,25(OH)2D) has anti-proliferative effects on several cancers. Vitamin D3 is synthesized by the enzyme, CYP27B1, and signals via the nuclear vitamin D3 receptor. The enzyme, CYP24A1, degrades vitamin D3. Aims (i) Measure the expression of CYP27B1, CYP24A1, and vitamin D3 receptor in human nonmalignant and cholangiocarcinoma lines and biopsy control or tumour samples; and (ii) evaluate the effects of vitamin D3 on vitamin D3 synthesis and cholangiocarcinoma growth. Methods In vitro studies were performed in malignant and nonmalignant cholangiocytes. Vitamin D3 receptor, CYP24 and CYP27 expression was measured in cell lines and biopsy samples. Cell lines were stimulated with vehicle or vitamin D3 from 30 min to 48 h. Cell viability was assessed by MTS assays and BrdU incorporation. Vitamin D3 receptor, CYP24A1 and CYP27B1 expression was measured in cholangiocarcinoma cells stimulated with vehicle or vitamin D3. Results In cholangiocarcinoma lines and biopsy samples, vitamin D3 receptor and CYP24A1 expression increased compared to controls, whereas CYP27B1 expression was decreased or unchanged. Vitamin D3 induced nuclear translocation of vitamin D3 receptor in cholangiocarcinoma and decreased cholangiocarcinoma growth. Conclusion Treatment with vitamin D3 decreased CYP24A1, whereas CYP27B1 expression increased. Modulation of vitamin D3 synthesis may be important in the management of cholangiocarcinoma.
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- 2013
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