Lysyl oxidase-like protein 2 (LOXL2) modulates barrier function in cholangiocytes in cholestasis

Marion J. Pollheimer, Silvia Racedo, Amanda Mikels-Vigdal, Derek Marshall, Christopher Bowlus, Carolin Lackner, Tobias Madl, Tom H. Karlsen, Johannes R. Hov, Susan K. Lyman, Joanne Adamkewicz, Victoria Smith, Emmanuel Moreau, Gernot Zollner, Tor Jacob Eide, Tatjana Stojakovic, Hubert Scharnagl, Hans Jürgen Gruber, Rudolf E. Stauber, Michael TraunerPeter Fickert

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background & Aims: The lysyl oxidase-like protein 2 (LOXL2) promotes stabilization of the extracellular matrix, chemotaxis, cell growth and cell mobility. We aimed to (i) identify stimuli of LOXL2 in cholangiopathies, (ii) characterize the effects of LOXL2 on biliary epithelial cells’ (BECs) barrier function, (iii) compare LOXL2 expression in primary sclerosing cholangitis (PSC), primary biliary cholangitis, and disease controls, and (iv) to determine LOXL2 expression and its cellular sources in four mouse models of cholangiopathies. Methods: Cultured murine BECs were challenged with well-known triggers of cellular senescence, hypoxia, phospholipid-deficient Abcb4−/− mouse bile and chenodeoxycholic acid and investigated for LOXL2, SNAIL1 and E-cadherin expression and transepithelial electrical resistance with and without LOX-inhibition. In vivo, LOXL2 expression was studied in PSC livers, and controls and mouse models. We compared LOXL2 serum levels in patients with PSC, secondary SC, primary biliary cholangitis, and controls. Results: Cellular senescence, hypoxia, Abcb4−/− bile and chenodeoxycholic acid induced LOXL2 and SNAIL1 expression, repressed E-cadherin expression, and significantly reduced transepithelial electrical resistance in BECs. Notably, all of the pathological changes could be recovered via pharmacological LOX-inhibition. Mouse models showed induced LOXL2 expression in the portal region and in association with ductular reaction. LOXL2 serum levels were significantly elevated in patients with cholangiopathies. In PSC, LOXL2 expression was located to characteristic periductal onion skin-type fibrosis, ductular reaction, Kupffer cells, and fibrotic septa. Importantly, in PSC, LOXL2 overexpression was paralleled by E-cadherin loss in BECs from medium-sized bile ducts. Conclusions: Reactive BECs produce LOXL2, resulting in increased tight junction permeability, which can be ameliorated by pharmacological LOX-inhibition in vitro. Reactive BECs, portal myofibroblasts, and Kupffer cells are the main sources of LOXL2 in cholangiopathies. Lay summary: In this study, we investigate the role of lysyl oxidase-like protein 2 (LOXL2), an enzyme pivotal in the development of organ fibrosis, in the pathogenesis of cholangiopathies (diseases of bile ducts), such as primary sclerosing cholangitis. We found LOXL2 to be expressed in association with bile duct epithelial injury and uncovered mechanisms for its upregulation and the subsequent effects in vitro and in vivo. Our findings support testing of anti-LOXL2 treatment strategies for patients with primary sclerosing cholangitis.

Original languageEnglish (US)
JournalJournal of Hepatology
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Protein-Lysine 6-Oxidase
Cholestasis
Sclerosing Cholangitis
Proteins
Epithelial Cells
Cadherins
Chenodeoxycholic Acid
Cell Hypoxia
Cholangitis
Kupffer Cells
Cell Aging
Bile Ducts
Bile Acids and Salts
Electric Impedance
Blood Proteins
Fibrosis
Bile Duct Diseases
Pharmacology

Keywords

  • Cholangiopathy
  • Epithelial barrier function
  • Liver fibrosis
  • Lysyl oxidases
  • Primary sclerosing cholangitis
  • Tight junction

ASJC Scopus subject areas

  • Hepatology

Cite this

Pollheimer, M. J., Racedo, S., Mikels-Vigdal, A., Marshall, D., Bowlus, C., Lackner, C., ... Fickert, P. (Accepted/In press). Lysyl oxidase-like protein 2 (LOXL2) modulates barrier function in cholangiocytes in cholestasis. Journal of Hepatology. https://doi.org/10.1016/j.jhep.2018.04.009

Lysyl oxidase-like protein 2 (LOXL2) modulates barrier function in cholangiocytes in cholestasis. / Pollheimer, Marion J.; Racedo, Silvia; Mikels-Vigdal, Amanda; Marshall, Derek; Bowlus, Christopher; Lackner, Carolin; Madl, Tobias; Karlsen, Tom H.; Hov, Johannes R.; Lyman, Susan K.; Adamkewicz, Joanne; Smith, Victoria; Moreau, Emmanuel; Zollner, Gernot; Eide, Tor Jacob; Stojakovic, Tatjana; Scharnagl, Hubert; Gruber, Hans Jürgen; Stauber, Rudolf E.; Trauner, Michael; Fickert, Peter.

In: Journal of Hepatology, 01.01.2018.

Research output: Contribution to journalArticle

Pollheimer, MJ, Racedo, S, Mikels-Vigdal, A, Marshall, D, Bowlus, C, Lackner, C, Madl, T, Karlsen, TH, Hov, JR, Lyman, SK, Adamkewicz, J, Smith, V, Moreau, E, Zollner, G, Eide, TJ, Stojakovic, T, Scharnagl, H, Gruber, HJ, Stauber, RE, Trauner, M & Fickert, P 2018, 'Lysyl oxidase-like protein 2 (LOXL2) modulates barrier function in cholangiocytes in cholestasis', Journal of Hepatology. https://doi.org/10.1016/j.jhep.2018.04.009
Pollheimer, Marion J. ; Racedo, Silvia ; Mikels-Vigdal, Amanda ; Marshall, Derek ; Bowlus, Christopher ; Lackner, Carolin ; Madl, Tobias ; Karlsen, Tom H. ; Hov, Johannes R. ; Lyman, Susan K. ; Adamkewicz, Joanne ; Smith, Victoria ; Moreau, Emmanuel ; Zollner, Gernot ; Eide, Tor Jacob ; Stojakovic, Tatjana ; Scharnagl, Hubert ; Gruber, Hans Jürgen ; Stauber, Rudolf E. ; Trauner, Michael ; Fickert, Peter. / Lysyl oxidase-like protein 2 (LOXL2) modulates barrier function in cholangiocytes in cholestasis. In: Journal of Hepatology. 2018.
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abstract = "Background & Aims: The lysyl oxidase-like protein 2 (LOXL2) promotes stabilization of the extracellular matrix, chemotaxis, cell growth and cell mobility. We aimed to (i) identify stimuli of LOXL2 in cholangiopathies, (ii) characterize the effects of LOXL2 on biliary epithelial cells’ (BECs) barrier function, (iii) compare LOXL2 expression in primary sclerosing cholangitis (PSC), primary biliary cholangitis, and disease controls, and (iv) to determine LOXL2 expression and its cellular sources in four mouse models of cholangiopathies. Methods: Cultured murine BECs were challenged with well-known triggers of cellular senescence, hypoxia, phospholipid-deficient Abcb4−/− mouse bile and chenodeoxycholic acid and investigated for LOXL2, SNAIL1 and E-cadherin expression and transepithelial electrical resistance with and without LOX-inhibition. In vivo, LOXL2 expression was studied in PSC livers, and controls and mouse models. We compared LOXL2 serum levels in patients with PSC, secondary SC, primary biliary cholangitis, and controls. Results: Cellular senescence, hypoxia, Abcb4−/− bile and chenodeoxycholic acid induced LOXL2 and SNAIL1 expression, repressed E-cadherin expression, and significantly reduced transepithelial electrical resistance in BECs. Notably, all of the pathological changes could be recovered via pharmacological LOX-inhibition. Mouse models showed induced LOXL2 expression in the portal region and in association with ductular reaction. LOXL2 serum levels were significantly elevated in patients with cholangiopathies. In PSC, LOXL2 expression was located to characteristic periductal onion skin-type fibrosis, ductular reaction, Kupffer cells, and fibrotic septa. Importantly, in PSC, LOXL2 overexpression was paralleled by E-cadherin loss in BECs from medium-sized bile ducts. Conclusions: Reactive BECs produce LOXL2, resulting in increased tight junction permeability, which can be ameliorated by pharmacological LOX-inhibition in vitro. Reactive BECs, portal myofibroblasts, and Kupffer cells are the main sources of LOXL2 in cholangiopathies. Lay summary: In this study, we investigate the role of lysyl oxidase-like protein 2 (LOXL2), an enzyme pivotal in the development of organ fibrosis, in the pathogenesis of cholangiopathies (diseases of bile ducts), such as primary sclerosing cholangitis. We found LOXL2 to be expressed in association with bile duct epithelial injury and uncovered mechanisms for its upregulation and the subsequent effects in vitro and in vivo. Our findings support testing of anti-LOXL2 treatment strategies for patients with primary sclerosing cholangitis.",
keywords = "Cholangiopathy, Epithelial barrier function, Liver fibrosis, Lysyl oxidases, Primary sclerosing cholangitis, Tight junction",
author = "Pollheimer, {Marion J.} and Silvia Racedo and Amanda Mikels-Vigdal and Derek Marshall and Christopher Bowlus and Carolin Lackner and Tobias Madl and Karlsen, {Tom H.} and Hov, {Johannes R.} and Lyman, {Susan K.} and Joanne Adamkewicz and Victoria Smith and Emmanuel Moreau and Gernot Zollner and Eide, {Tor Jacob} and Tatjana Stojakovic and Hubert Scharnagl and Gruber, {Hans J{\"u}rgen} and Stauber, {Rudolf E.} and Michael Trauner and Peter Fickert",
year = "2018",
month = "1",
day = "1",
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TY - JOUR

T1 - Lysyl oxidase-like protein 2 (LOXL2) modulates barrier function in cholangiocytes in cholestasis

AU - Pollheimer, Marion J.

AU - Racedo, Silvia

AU - Mikels-Vigdal, Amanda

AU - Marshall, Derek

AU - Bowlus, Christopher

AU - Lackner, Carolin

AU - Madl, Tobias

AU - Karlsen, Tom H.

AU - Hov, Johannes R.

AU - Lyman, Susan K.

AU - Adamkewicz, Joanne

AU - Smith, Victoria

AU - Moreau, Emmanuel

AU - Zollner, Gernot

AU - Eide, Tor Jacob

AU - Stojakovic, Tatjana

AU - Scharnagl, Hubert

AU - Gruber, Hans Jürgen

AU - Stauber, Rudolf E.

AU - Trauner, Michael

AU - Fickert, Peter

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Background & Aims: The lysyl oxidase-like protein 2 (LOXL2) promotes stabilization of the extracellular matrix, chemotaxis, cell growth and cell mobility. We aimed to (i) identify stimuli of LOXL2 in cholangiopathies, (ii) characterize the effects of LOXL2 on biliary epithelial cells’ (BECs) barrier function, (iii) compare LOXL2 expression in primary sclerosing cholangitis (PSC), primary biliary cholangitis, and disease controls, and (iv) to determine LOXL2 expression and its cellular sources in four mouse models of cholangiopathies. Methods: Cultured murine BECs were challenged with well-known triggers of cellular senescence, hypoxia, phospholipid-deficient Abcb4−/− mouse bile and chenodeoxycholic acid and investigated for LOXL2, SNAIL1 and E-cadherin expression and transepithelial electrical resistance with and without LOX-inhibition. In vivo, LOXL2 expression was studied in PSC livers, and controls and mouse models. We compared LOXL2 serum levels in patients with PSC, secondary SC, primary biliary cholangitis, and controls. Results: Cellular senescence, hypoxia, Abcb4−/− bile and chenodeoxycholic acid induced LOXL2 and SNAIL1 expression, repressed E-cadherin expression, and significantly reduced transepithelial electrical resistance in BECs. Notably, all of the pathological changes could be recovered via pharmacological LOX-inhibition. Mouse models showed induced LOXL2 expression in the portal region and in association with ductular reaction. LOXL2 serum levels were significantly elevated in patients with cholangiopathies. In PSC, LOXL2 expression was located to characteristic periductal onion skin-type fibrosis, ductular reaction, Kupffer cells, and fibrotic septa. Importantly, in PSC, LOXL2 overexpression was paralleled by E-cadherin loss in BECs from medium-sized bile ducts. Conclusions: Reactive BECs produce LOXL2, resulting in increased tight junction permeability, which can be ameliorated by pharmacological LOX-inhibition in vitro. Reactive BECs, portal myofibroblasts, and Kupffer cells are the main sources of LOXL2 in cholangiopathies. Lay summary: In this study, we investigate the role of lysyl oxidase-like protein 2 (LOXL2), an enzyme pivotal in the development of organ fibrosis, in the pathogenesis of cholangiopathies (diseases of bile ducts), such as primary sclerosing cholangitis. We found LOXL2 to be expressed in association with bile duct epithelial injury and uncovered mechanisms for its upregulation and the subsequent effects in vitro and in vivo. Our findings support testing of anti-LOXL2 treatment strategies for patients with primary sclerosing cholangitis.

AB - Background & Aims: The lysyl oxidase-like protein 2 (LOXL2) promotes stabilization of the extracellular matrix, chemotaxis, cell growth and cell mobility. We aimed to (i) identify stimuli of LOXL2 in cholangiopathies, (ii) characterize the effects of LOXL2 on biliary epithelial cells’ (BECs) barrier function, (iii) compare LOXL2 expression in primary sclerosing cholangitis (PSC), primary biliary cholangitis, and disease controls, and (iv) to determine LOXL2 expression and its cellular sources in four mouse models of cholangiopathies. Methods: Cultured murine BECs were challenged with well-known triggers of cellular senescence, hypoxia, phospholipid-deficient Abcb4−/− mouse bile and chenodeoxycholic acid and investigated for LOXL2, SNAIL1 and E-cadherin expression and transepithelial electrical resistance with and without LOX-inhibition. In vivo, LOXL2 expression was studied in PSC livers, and controls and mouse models. We compared LOXL2 serum levels in patients with PSC, secondary SC, primary biliary cholangitis, and controls. Results: Cellular senescence, hypoxia, Abcb4−/− bile and chenodeoxycholic acid induced LOXL2 and SNAIL1 expression, repressed E-cadherin expression, and significantly reduced transepithelial electrical resistance in BECs. Notably, all of the pathological changes could be recovered via pharmacological LOX-inhibition. Mouse models showed induced LOXL2 expression in the portal region and in association with ductular reaction. LOXL2 serum levels were significantly elevated in patients with cholangiopathies. In PSC, LOXL2 expression was located to characteristic periductal onion skin-type fibrosis, ductular reaction, Kupffer cells, and fibrotic septa. Importantly, in PSC, LOXL2 overexpression was paralleled by E-cadherin loss in BECs from medium-sized bile ducts. Conclusions: Reactive BECs produce LOXL2, resulting in increased tight junction permeability, which can be ameliorated by pharmacological LOX-inhibition in vitro. Reactive BECs, portal myofibroblasts, and Kupffer cells are the main sources of LOXL2 in cholangiopathies. Lay summary: In this study, we investigate the role of lysyl oxidase-like protein 2 (LOXL2), an enzyme pivotal in the development of organ fibrosis, in the pathogenesis of cholangiopathies (diseases of bile ducts), such as primary sclerosing cholangitis. We found LOXL2 to be expressed in association with bile duct epithelial injury and uncovered mechanisms for its upregulation and the subsequent effects in vitro and in vivo. Our findings support testing of anti-LOXL2 treatment strategies for patients with primary sclerosing cholangitis.

KW - Cholangiopathy

KW - Epithelial barrier function

KW - Liver fibrosis

KW - Lysyl oxidases

KW - Primary sclerosing cholangitis

KW - Tight junction

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