Hedgehog signalling stimulates precursor cell accumulation and impairs epithelial maturation in the murine oesophagus
- Willemijn A van Dop1,2,
- Sanne L Rosekrans1,2,
- Anja Uhmann3,
- Viljar Jaks4,
- G Johan A Offerhaus5,6,
- Marius A van den Bergh Weerman6,
- Maria Kasper4,
- Jarom Heijmans1,2,
- James C H Hardwick7,
- Hein W Verspaget7,
- Daan W Hommes7,8,
- Rune Toftgård4,
- Heidi Hahn3,
- Gijs R van den Brink1,2
- 1Tytgat Institute for Liver and Intestinal Research, Academic Medical Center, Amsterdam, The Netherlands
- 2Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands
- 3Institute of Human Genetics, University Medical Center Göttingen, Göttingen, Germany
- 4Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden
- 5Department of Pathology, University Medical Center Utrecht, Utrecht, The Netherlands
- 6Department of Pathology, Academic Medical Center, Amsterdam, The Netherlands
- 7Department of Gastroenterology and Hepatology, Leiden University Medical Center, Leiden, The Netherlands
- 8Center for Inflammatory Bowel Diseases, University of California Los Angeles, Los Angeles, USA
- Correspondence to Dr Gijs R van den Brink, Tytgat Institute for Liver and Intestinal Research, Academic Medical Center, Amsterdam 1105 AZ, The Netherlands;
Contributors WAvD designed and performed experiments, analysed data and wrote the manuscript. SLR designed and performed experiments, analysed data and wrote the manuscript. AU and HH provided the Ptch1flox/floxCreERT2+/− mice and commented on the manuscript. VJ, RT and MK provided the K5tTA-TreGli1 mice and commented on the manuscript. GJAO helped in judging the histology of the tissue sections. MAvdBW performed the EM. JH designed and performed experiments, analysed data and commented on the manuscript. HWV provided the human tissue samples. DWH and JCHH advised on experimental design and commented on the manuscript. GRvdB designed experiments, analysed data, wrote the manuscript and supervised and mentored all work. WAvD and SLR contributed equally to this work.
- Revised 20 February 2012
- Accepted 22 February 2012
- Published Online First 13 April 2012
Objective In the intestine Hedgehog (Hh) signalling is directed from epithelium to mesenchyme and negatively regulates epithelial precursor cell fate. The role of Hh signalling in the oesophagus has not been studied in vivo. Here the authors examined the role of Hh signalling in epithelial homeostasis of oesophagus.
Design The authors used transgenic mice in which the Hh receptor Patched1 (Ptch1) could be conditionally inactivated in a body-wide manner and mice in which Gli1 could be induced specifically in the epithelium of the skin and oesophagus. Effects on epithelial homeostasis of the oesophagus were examined using immunohistochemistry, in situ hybridisation, transmission electron microscopy and real-time PCR. Hh signalling was examined in patients with oesophageal squamous cell carcinoma (SCC) by quantitative real-time PCR.
Results Sonic Hh is signalled in an autocrine manner in the basal layer of the oesophagus. Activation of Hh signalling resulted in an expansion of the epithelial precursor cell compartment and failure of epithelial maturation and migration. Levels of Hh targets GLI1, HHIP and PTCH1 were increased in SCC compared with normal tissue from the same patients.
Conclusion Here the authors find that Hh signalling positively regulates the precursor cell compartment in the oesophageal epithelium in an autocrine manner. Since Hh signalling targets precursor cells in the oesophageal epithelium and signalling is increased in SCCs, Hh signalling may be involved in oesophageal SCC formation.
- Hedgehog proteins
- stem cells
- oesophageal cancer
- colon carcinogenesis
- colorectal carcinoma
- colorectal cancer screening
- colorectal adenomas
- molecular oncology
- inflammatory bowel disease
- oxidative metabolism
- matrix metalloproteinase
- colonic adenomas
- hedgehog signalling
- signal transduction
Funding This study was supported by a VIDI grant from The Netherlands Organization for Scientific Research, from the European Research Council under the European Community's Seventh Framework Program (FP7/2007-2013)/ERC grant agreement number 241344, by a grant from the Dutch Digestive Foundation and by a grant of the Deutsche Forschungsgemeinschaft HA 2197/5-1 FOR942.
Competing interests None.
Patient consent Twenty patients with oesophageal squamous cell carcinoma were selected from a tissue biobank of the Leiden University Medical Center (LUMC). Tissue stored in the biobank has been collected from excess material from patients operated for gastrointestinal cancer. The study on human material was performed according to the instructions and guidelines of the LUMC Medical Ethics Committee and in compliance with the Helsinki Declaration.
Ethics approval The ethics approval was provided by LUMC Medical Ethics Committee.
Provenance and peer review Not commissioned; externally peer reviewed.