Widespread GLI expression but limited canonical hedgehog signaling restricted to the ductular reaction in human chronic liver disease

Canonical Hedgehog (Hh) signaling in vertebrate cells occurs following Smoothened activation/translocation into the primary cilia (Pc), followed by a GLI transcriptional response. Nonetheless, GLI activation can occur independently of the canonical Hh pathway. Using a murine model of liver injury, w...

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Main Authors: Grzelak, C., Sigglekow, N., Tirnitz-Parker, Nina, Hamson, E., Warren, A., Maneck, B., Chen, J., Patkunanathan, B., Boland, J., Cheng, R., Shackel, N., Seth, D., Bowen, D., Martelotto, L., Watkins, D., McCaughan, G.
Format: Journal Article
Published: Public Library of Science 2017
Online Access:http://hdl.handle.net/20.500.11937/50290
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author Grzelak, C.
Sigglekow, N.
Tirnitz-Parker, Nina
Hamson, E.
Warren, A.
Maneck, B.
Chen, J.
Patkunanathan, B.
Boland, J.
Cheng, R.
Shackel, N.
Seth, D.
Bowen, D.
Martelotto, L.
Watkins, D.
McCaughan, G.
author_facet Grzelak, C.
Sigglekow, N.
Tirnitz-Parker, Nina
Hamson, E.
Warren, A.
Maneck, B.
Chen, J.
Patkunanathan, B.
Boland, J.
Cheng, R.
Shackel, N.
Seth, D.
Bowen, D.
Martelotto, L.
Watkins, D.
McCaughan, G.
author_sort Grzelak, C.
building Curtin Institutional Repository
collection Online Access
description Canonical Hedgehog (Hh) signaling in vertebrate cells occurs following Smoothened activation/translocation into the primary cilia (Pc), followed by a GLI transcriptional response. Nonetheless, GLI activation can occur independently of the canonical Hh pathway. Using a murine model of liver injury, we previously identified the importance of canonical Hh signaling within the Pc+ liver progenitor cell (LPC) population and noted that SMO-independent, GLI-mediated signals were important in multiple Pc-ve GLI2+ intrahepatic populations. This study extends these observations to human liver tissue, and analyses the effect of GLI inhibition on LPC viability/gene expression. Human donor and cirrhotic liver tissue specimens were evaluated for SHH, GLI2 and Pc expression using immunofluorescence and qRT-PCR. Changes to viability and gene expression in LPCs in vitro were assessed following GLI inhibition. Identification of Pc (as a marker of canonical Hh signaling) in human cirrhosis was predominantly confined to the ductular reaction and LPCs. In contrast, GLI2 was expressed in multiple cell populations including Pc-ve endothelium, hepatocytes, and leukocytes. HSCs/myofibroblasts (gt;99%) expressed GLI2, with only 1.92% displaying Pc. In vitro GLI signals maintained proliferation/viability within LPCs and GLI inhibition affected the expression of genes related to stemness, hepatocyte/biliary differentiation and Hh/Wnt signaling. At least two mechanisms of GLI signaling (Pc/SMOdependent and Pc/SMO-independent) mediate chronic liver disease pathogenesis. This may have significant ramifications for the choice of Hh inhibitor (anti-SMO or anti-GLI) suitable for clinical trials. We also postulate GLI delivers a pro-survival signal to LPCs whilst maintaining stemness.
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institution Curtin University Malaysia
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spelling curtin-20.500.11937-502902017-09-13T15:37:23Z Widespread GLI expression but limited canonical hedgehog signaling restricted to the ductular reaction in human chronic liver disease Grzelak, C. Sigglekow, N. Tirnitz-Parker, Nina Hamson, E. Warren, A. Maneck, B. Chen, J. Patkunanathan, B. Boland, J. Cheng, R. Shackel, N. Seth, D. Bowen, D. Martelotto, L. Watkins, D. McCaughan, G. Canonical Hedgehog (Hh) signaling in vertebrate cells occurs following Smoothened activation/translocation into the primary cilia (Pc), followed by a GLI transcriptional response. Nonetheless, GLI activation can occur independently of the canonical Hh pathway. Using a murine model of liver injury, we previously identified the importance of canonical Hh signaling within the Pc+ liver progenitor cell (LPC) population and noted that SMO-independent, GLI-mediated signals were important in multiple Pc-ve GLI2+ intrahepatic populations. This study extends these observations to human liver tissue, and analyses the effect of GLI inhibition on LPC viability/gene expression. Human donor and cirrhotic liver tissue specimens were evaluated for SHH, GLI2 and Pc expression using immunofluorescence and qRT-PCR. Changes to viability and gene expression in LPCs in vitro were assessed following GLI inhibition. Identification of Pc (as a marker of canonical Hh signaling) in human cirrhosis was predominantly confined to the ductular reaction and LPCs. In contrast, GLI2 was expressed in multiple cell populations including Pc-ve endothelium, hepatocytes, and leukocytes. HSCs/myofibroblasts (gt;99%) expressed GLI2, with only 1.92% displaying Pc. In vitro GLI signals maintained proliferation/viability within LPCs and GLI inhibition affected the expression of genes related to stemness, hepatocyte/biliary differentiation and Hh/Wnt signaling. At least two mechanisms of GLI signaling (Pc/SMOdependent and Pc/SMO-independent) mediate chronic liver disease pathogenesis. This may have significant ramifications for the choice of Hh inhibitor (anti-SMO or anti-GLI) suitable for clinical trials. We also postulate GLI delivers a pro-survival signal to LPCs whilst maintaining stemness. 2017 Journal Article http://hdl.handle.net/20.500.11937/50290 10.1371/journal.pone.0171480 https://creativecommons.org/publicdomain/zero/1.0/ Public Library of Science fulltext
spellingShingle Grzelak, C.
Sigglekow, N.
Tirnitz-Parker, Nina
Hamson, E.
Warren, A.
Maneck, B.
Chen, J.
Patkunanathan, B.
Boland, J.
Cheng, R.
Shackel, N.
Seth, D.
Bowen, D.
Martelotto, L.
Watkins, D.
McCaughan, G.
Widespread GLI expression but limited canonical hedgehog signaling restricted to the ductular reaction in human chronic liver disease
title Widespread GLI expression but limited canonical hedgehog signaling restricted to the ductular reaction in human chronic liver disease
title_full Widespread GLI expression but limited canonical hedgehog signaling restricted to the ductular reaction in human chronic liver disease
title_fullStr Widespread GLI expression but limited canonical hedgehog signaling restricted to the ductular reaction in human chronic liver disease
title_full_unstemmed Widespread GLI expression but limited canonical hedgehog signaling restricted to the ductular reaction in human chronic liver disease
title_short Widespread GLI expression but limited canonical hedgehog signaling restricted to the ductular reaction in human chronic liver disease
title_sort widespread gli expression but limited canonical hedgehog signaling restricted to the ductular reaction in human chronic liver disease
url http://hdl.handle.net/20.500.11937/50290