Taurocholate Induces Biliary Differentiation of Liver Progenitor Cells Causing Hepatic Stellate Cell Chemotaxis in the Ductular Reaction: Role in Pediatric Cystic Fibrosis Liver Disease

Katarzyna N. Pozniak, Michael A. Pearen, Tamara N. Pereira, Cynthia S.M. Kramer, Priyakshi Kalita-De Croft, Sujeevi K. Nawaratna, Manuel A. Fernandez-Rojo, Geoffrey N. Gobert, Janina E.E. Tirnitz-Parker, John K. Olynyk, Ross W. Shepherd, Peter J. Lewindon, Grant A. Ramm

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Abstract

Cystic fibrosis liver disease (CFLD) in children causes progressive fibrosis leading to biliary cirrhosis; however, its cause(s) and early pathogenesis are unclear. We hypothesized that a bile acid–induced ductular reaction (DR) drives fibrogenesis. The DR was evaluated by cytokeratin-7 immunohistochemistry in liver biopsies, staged for fibrosis, from 60 children with CFLD, and it demonstrated that the DR was significantly correlated with hepatic fibrosis stage and biliary taurocholate levels. To examine the mechanisms involved in DR induction, liver progenitor cells (LPCs) were treated with taurocholate, and key events in DR evolution were assessed: LPC proliferation, LPC biliary differentiation, and hepatic stellate cell (HSC) chemotaxis. Taurocholate induced a time-dependent increase in LPC proliferation and expression of genes associated with cholangiocyte differentiation (cytokeratin 19, connexin 43, integrin β4, and γ-glutamyltranspeptidase), whereas the hepatocyte specification marker HNF4α was suppressed. Functional cholangiocyte differentiation was demonstrated via increased acetylated α-tubulin and SOX9 proteins, the number of primary cilia+ LPCs, and increased active γ-glutamyltranspeptidase enzyme secretion. Taurocholate induced LPCs to release MCP-1, MIP1α, and RANTES into conditioned medium causing HSC chemotaxis, which was inhibited by anti-MIP1α. Immunofluorescence confirmed chemokine expression localized to CK7+ DR and LPCs in CFLD liver biopsies. This study suggests that taurocholate is involved in initiating functional LPC biliary differentiation and the development of the DR, with subsequent induction of chemokines that drive HSC recruitment in CFLD.

Original languageEnglish
Pages (from-to)2744-2757
Number of pages14
JournalAmerican Journal of Pathology
Volume187
Issue number12
DOIs
Publication statusPublished - 1 Dec 2017

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Hepatic Stellate Cells
Taurocholic Acid
Chemotaxis
Cystic Fibrosis
Liver Diseases
Stem Cells
Pediatrics
Liver
Fibrosis
Chemokines
Cell Differentiation
Cell Proliferation
Keratin-7
Keratin-19
Biopsy
Chemokine CCL5
Connexin 43
Biliary Liver Cirrhosis
Cilia
Tubulin

Cite this

Pozniak, Katarzyna N. ; Pearen, Michael A. ; Pereira, Tamara N. ; Kramer, Cynthia S.M. ; Kalita-De Croft, Priyakshi ; Nawaratna, Sujeevi K. ; Fernandez-Rojo, Manuel A. ; Gobert, Geoffrey N. ; Tirnitz-Parker, Janina E.E. ; Olynyk, John K. ; Shepherd, Ross W. ; Lewindon, Peter J. ; Ramm, Grant A. / Taurocholate Induces Biliary Differentiation of Liver Progenitor Cells Causing Hepatic Stellate Cell Chemotaxis in the Ductular Reaction : Role in Pediatric Cystic Fibrosis Liver Disease. In: American Journal of Pathology. 2017 ; Vol. 187, No. 12. pp. 2744-2757.
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abstract = "Cystic fibrosis liver disease (CFLD) in children causes progressive fibrosis leading to biliary cirrhosis; however, its cause(s) and early pathogenesis are unclear. We hypothesized that a bile acid–induced ductular reaction (DR) drives fibrogenesis. The DR was evaluated by cytokeratin-7 immunohistochemistry in liver biopsies, staged for fibrosis, from 60 children with CFLD, and it demonstrated that the DR was significantly correlated with hepatic fibrosis stage and biliary taurocholate levels. To examine the mechanisms involved in DR induction, liver progenitor cells (LPCs) were treated with taurocholate, and key events in DR evolution were assessed: LPC proliferation, LPC biliary differentiation, and hepatic stellate cell (HSC) chemotaxis. Taurocholate induced a time-dependent increase in LPC proliferation and expression of genes associated with cholangiocyte differentiation (cytokeratin 19, connexin 43, integrin β4, and γ-glutamyltranspeptidase), whereas the hepatocyte specification marker HNF4α was suppressed. Functional cholangiocyte differentiation was demonstrated via increased acetylated α-tubulin and SOX9 proteins, the number of primary cilia+ LPCs, and increased active γ-glutamyltranspeptidase enzyme secretion. Taurocholate induced LPCs to release MCP-1, MIP1α, and RANTES into conditioned medium causing HSC chemotaxis, which was inhibited by anti-MIP1α. Immunofluorescence confirmed chemokine expression localized to CK7+ DR and LPCs in CFLD liver biopsies. This study suggests that taurocholate is involved in initiating functional LPC biliary differentiation and the development of the DR, with subsequent induction of chemokines that drive HSC recruitment in CFLD.",
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Pozniak, KN, Pearen, MA, Pereira, TN, Kramer, CSM, Kalita-De Croft, P, Nawaratna, SK, Fernandez-Rojo, MA, Gobert, GN, Tirnitz-Parker, JEE, Olynyk, JK, Shepherd, RW, Lewindon, PJ & Ramm, GA 2017, 'Taurocholate Induces Biliary Differentiation of Liver Progenitor Cells Causing Hepatic Stellate Cell Chemotaxis in the Ductular Reaction: Role in Pediatric Cystic Fibrosis Liver Disease' American Journal of Pathology, vol. 187, no. 12, pp. 2744-2757. https://doi.org/10.1016/j.ajpath.2017.08.024

Taurocholate Induces Biliary Differentiation of Liver Progenitor Cells Causing Hepatic Stellate Cell Chemotaxis in the Ductular Reaction : Role in Pediatric Cystic Fibrosis Liver Disease. / Pozniak, Katarzyna N.; Pearen, Michael A.; Pereira, Tamara N.; Kramer, Cynthia S.M.; Kalita-De Croft, Priyakshi; Nawaratna, Sujeevi K.; Fernandez-Rojo, Manuel A.; Gobert, Geoffrey N.; Tirnitz-Parker, Janina E.E.; Olynyk, John K.; Shepherd, Ross W.; Lewindon, Peter J.; Ramm, Grant A.

In: American Journal of Pathology, Vol. 187, No. 12, 01.12.2017, p. 2744-2757.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Taurocholate Induces Biliary Differentiation of Liver Progenitor Cells Causing Hepatic Stellate Cell Chemotaxis in the Ductular Reaction

T2 - Role in Pediatric Cystic Fibrosis Liver Disease

AU - Pozniak, Katarzyna N.

AU - Pearen, Michael A.

AU - Pereira, Tamara N.

AU - Kramer, Cynthia S.M.

AU - Kalita-De Croft, Priyakshi

AU - Nawaratna, Sujeevi K.

AU - Fernandez-Rojo, Manuel A.

AU - Gobert, Geoffrey N.

AU - Tirnitz-Parker, Janina E.E.

AU - Olynyk, John K.

AU - Shepherd, Ross W.

AU - Lewindon, Peter J.

AU - Ramm, Grant A.

PY - 2017/12/1

Y1 - 2017/12/1

N2 - Cystic fibrosis liver disease (CFLD) in children causes progressive fibrosis leading to biliary cirrhosis; however, its cause(s) and early pathogenesis are unclear. We hypothesized that a bile acid–induced ductular reaction (DR) drives fibrogenesis. The DR was evaluated by cytokeratin-7 immunohistochemistry in liver biopsies, staged for fibrosis, from 60 children with CFLD, and it demonstrated that the DR was significantly correlated with hepatic fibrosis stage and biliary taurocholate levels. To examine the mechanisms involved in DR induction, liver progenitor cells (LPCs) were treated with taurocholate, and key events in DR evolution were assessed: LPC proliferation, LPC biliary differentiation, and hepatic stellate cell (HSC) chemotaxis. Taurocholate induced a time-dependent increase in LPC proliferation and expression of genes associated with cholangiocyte differentiation (cytokeratin 19, connexin 43, integrin β4, and γ-glutamyltranspeptidase), whereas the hepatocyte specification marker HNF4α was suppressed. Functional cholangiocyte differentiation was demonstrated via increased acetylated α-tubulin and SOX9 proteins, the number of primary cilia+ LPCs, and increased active γ-glutamyltranspeptidase enzyme secretion. Taurocholate induced LPCs to release MCP-1, MIP1α, and RANTES into conditioned medium causing HSC chemotaxis, which was inhibited by anti-MIP1α. Immunofluorescence confirmed chemokine expression localized to CK7+ DR and LPCs in CFLD liver biopsies. This study suggests that taurocholate is involved in initiating functional LPC biliary differentiation and the development of the DR, with subsequent induction of chemokines that drive HSC recruitment in CFLD.

AB - Cystic fibrosis liver disease (CFLD) in children causes progressive fibrosis leading to biliary cirrhosis; however, its cause(s) and early pathogenesis are unclear. We hypothesized that a bile acid–induced ductular reaction (DR) drives fibrogenesis. The DR was evaluated by cytokeratin-7 immunohistochemistry in liver biopsies, staged for fibrosis, from 60 children with CFLD, and it demonstrated that the DR was significantly correlated with hepatic fibrosis stage and biliary taurocholate levels. To examine the mechanisms involved in DR induction, liver progenitor cells (LPCs) were treated with taurocholate, and key events in DR evolution were assessed: LPC proliferation, LPC biliary differentiation, and hepatic stellate cell (HSC) chemotaxis. Taurocholate induced a time-dependent increase in LPC proliferation and expression of genes associated with cholangiocyte differentiation (cytokeratin 19, connexin 43, integrin β4, and γ-glutamyltranspeptidase), whereas the hepatocyte specification marker HNF4α was suppressed. Functional cholangiocyte differentiation was demonstrated via increased acetylated α-tubulin and SOX9 proteins, the number of primary cilia+ LPCs, and increased active γ-glutamyltranspeptidase enzyme secretion. Taurocholate induced LPCs to release MCP-1, MIP1α, and RANTES into conditioned medium causing HSC chemotaxis, which was inhibited by anti-MIP1α. Immunofluorescence confirmed chemokine expression localized to CK7+ DR and LPCs in CFLD liver biopsies. This study suggests that taurocholate is involved in initiating functional LPC biliary differentiation and the development of the DR, with subsequent induction of chemokines that drive HSC recruitment in CFLD.

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U2 - 10.1016/j.ajpath.2017.08.024

DO - 10.1016/j.ajpath.2017.08.024

M3 - Article

VL - 187

SP - 2744

EP - 2757

JO - The American Journal of Pathology

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