title: Stromal Indian hedgehog signaling is required for intestinal adenoma formation in mice.
abstract: OBJECTIVE
  Indian hedgehog (IHH) is an epithelial-derived signal in the intestinal stroma, inducing factors that restrict epithelial proliferation and suppress activation of the immune system. In addition to these rapid effects of IHH signaling, IHH is required to maintain a stromal phenotype in which myofibroblasts and smooth muscle cells predominate. We investigated the role of IHH signaling during development of intestinal neoplasia in mice.
  
  
  METHODS
  Glioma-associated oncogene (Gli1)-CreERT2 and Patched (Ptch)-lacZ reporter mice were crossed with Apc(Min) mice to generate Gli1CreERT2-Rosa26-ZSGreen-Apc(Min) and Ptch-lacZ-Apc(Min) mice, which were used to identify hedgehog-responsive cells. Cyp1a1Cre-Apc (Apc(HET)) mice, which develop adenomas after administration of β-naphthoflavone, were crossed with mice with conditional disruption of Ihh in the small intestine epithelium. Apc(Min) mice were crossed with mice in which sonic hedgehog (SHH) was overexpressed specifically in the intestinal epithelium. Intestinal tissues were collected and analyzed histologically and by immunohistochemistry and quantitative reverse-transcription polymerase chain reaction. We also analyzed levels of IHH messenger RNA and expression of IHH gene targets in intestinal tissues from patients with familial adenomatous polyposis (n = 18) or sessile serrated adenomas (n = 15) and normal colonic tissue from control patients (n = 12).
  
  
  RESULTS
  Expression of IHH messenger RNA and its targets were increased in intestinal adenomas from patients and mice compared with control colon tissues. In mice, IHH signaling was exclusively paracrine, from the epithelium to the stroma. Loss of IHH from Apc(HET) mice almost completely blocked adenoma development, and overexpression of SHH increased the number and size of adenomas that developed. Loss of IHH from Apc(HET) mice changed the composition of the adenoma stroma; cells that expressed α-smooth muscle actin or desmin were lost, along with expression of cyclooxygenase-2, and the number of vimentin-positive cells increased.
  
  
  CONCLUSIONS
  Apc mutant epithelial cells secrete IHH to maintain an intestinal stromal phenotype that is required for adenoma development in mice.
authors.affiliation: Tytgat Institute for Liver and Intestinal Research and Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Nikè V J A
authors.initials: NV
authors.lastname: Büller
authors.affiliation: Tytgat Institute for Liver and Intestinal Research and Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Sanne L
authors.initials: SL
authors.lastname: Rosekrans
authors.affiliation: Molecular Oncology Department, Genentech, South San Francisco, California.
authors.forename: Ciara
authors.initials: C
authors.lastname: Metcalfe
authors.affiliation: Tytgat Institute for Liver and Intestinal Research and Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Jarom
authors.initials: J
authors.lastname: Heijmans
authors.affiliation: Tytgat Institute for Liver and Intestinal Research and Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Willemijn A
authors.initials: WA
authors.lastname: van Dop
authors.affiliation: Laboratory for Experimental Oncology and Radiobiology (LEXOR), Center for Experimental and Molecular Medicine, Academic Medical Center (AMC), Amsterdam, The Netherlands.
authors.forename: Evelyn
authors.initials: E
authors.lastname: Fessler
authors.affiliation: Barts Cancer Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK.
authors.forename: Marnix
authors.initials: M
authors.lastname: Jansen
authors.affiliation: Molecular Oncology Department, Genentech, South San Francisco, California.
authors.forename: Christina
authors.initials: C
authors.lastname: Ahn
authors.affiliation: Tytgat Institute for Liver and Intestinal Research and Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Jacqueline L M
authors.initials: JL
authors.lastname: Vermeulen
authors.affiliation: Tytgat Institute for Liver and Intestinal Research and Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: B Florien
authors.initials: BF
authors.lastname: Westendorp
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Els C
authors.initials: EC
authors.lastname: Robanus-Maandag
authors.affiliation: Department of Pathology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: G Johan
authors.initials: GJ
authors.lastname: Offerhaus
authors.affiliation: Laboratory for Experimental Oncology and Radiobiology (LEXOR), Center for Experimental and Molecular Medicine, Academic Medical Center (AMC), Amsterdam, The Netherlands.
authors.forename: Jan Paul
authors.initials: JP
authors.lastname: Medema
authors.affiliation: Tytgat Institute for Liver and Intestinal Research and Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Geert R A M
authors.initials: GR
authors.lastname: D'Haens
authors.affiliation: Tytgat Institute for Liver and Intestinal Research and Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Manon E
authors.initials: ME
authors.lastname: Wildenberg
authors.affiliation: Molecular Oncology Department, Genentech, South San Francisco, California.
authors.forename: Frederic J
authors.initials: FJ
authors.lastname: de Sauvage
authors.affiliation: Tytgat Institute for Liver and Intestinal Research and Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Vanesa
authors.initials: V
authors.lastname: Muncan
authors.affiliation: Tytgat Institute for Liver and Intestinal Research and Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands. Electronic address: g.r.vandenbrink@amc.nl.
authors.forename: Gijs R
authors.initials: GR
authors.lastname: van den Brink
mesh_terms: D000236:Adenoma / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D011125:Adenomatous Polyposis Coli / Q000378:metabolism / Q000473:pathology; D000818:Animals; D019898:Autocrine Communication; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D016022:Case-Control Studies; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000378:metabolism / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D017491:Genes, APC; D020022:Genetic Predisposition to Disease; D053823:Hedgehog Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006965:Hyperplasia; D019426:Integrases / Q000235:genetics; D007413:Intestinal Mucosa / Q000378:metabolism / Q000473:pathology; D007414:Intestinal Neoplasms / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008822:Mice, Transgenic; D009154:Mutation; D019899:Paracrine Communication; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D017154:Stromal Cells / Q000378:metabolism* / Q000473:pathology; D047368:Tumor Burden; D019324:beta-Naphthoflavone
chemical_list: D014408:Biomarkers, Tumor; D053823:Hedgehog Proteins; C509246:IHH protein, human; D012333:RNA, Messenger; C465570:Shh protein, mouse; C514061:ihh protein, mouse; D019324:beta-Naphthoflavone; D019363:Cytochrome P-450 CYP1A1; C045073:Cre recombinase; D019426:Integrases
keywords: COX2; Colon Cancer; Mouse Model; Prostaglandin