title: Association of hepatocyte-derived growth factor receptor/caudal type homeobox 2 co-expression with mucosal regeneration in active ulcerative colitis.
abstract: OBJECTIVE
  To characterize the regeneration-associated stem cell-related phenotype of hepatocyte-derived growth factor receptor (HGFR)-expressing cells in active ulcerative colitis (UC).
  
  
  METHODS
  On the whole 38 peripheral blood samples and 38 colonic biopsy samples from 18 patients with histologically proven active UC and 20 healthy control subjects were collected. After preparing tissue microarrays and blood smears HGFR, caudal type homeobox 2 (CDX2), prominin-1 (CD133) and Musashi-1 conventional and double fluorescent immunolabelings were performed. Immunostained samples were digitalized using high-resolution Mirax Desk instrument, and analyzed with the Mirax TMA Module software. For semiquantitative counting of immunopositive lamina propria (LP) cells 5 fields of view were counted at magnification × 200 in each sample core, then mean ± SD were determined. In case of peripheral blood smears, 30 fields of view with 100 μm diameter were evaluated in every sample and the number of immunopositive cells (mean ± SD) was determined. Using 337 nm UVA Laser MicroDissection system at least 5000 subepithelial cells from the lamina propria were collected. Gene expression analysis of HGFR, CDX2, CD133, leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5), Musashi-1 and cytokeratin 20 (CK20) were performed in both laser-microdisscted samples and blood samples by using real time reverse transcription polymerase chain reaction (RT-PCR).
  
  
  RESULTS
  By performing conventional and double fluorescent immunolabelings confirmed by RT-PCR, higher number of HGFR (blood: 6.7 ± 1.22 vs 38.5 ± 3.18; LP: 2.25 ± 0.85 vs 9.22 ± 0.65; P < 0.05), CDX2 (blood: 0 vs 0.94 ± 0.64; LP: 0.75 ± 0.55 vs 2.11 ± 0.75; P < 0.05), CD133 (blood: 1.1 ± 0.72 vs 8.3 ± 1.08; LP: 11.1 ± 0.85 vs 26.28 ± 1.71; P < 0.05) and Musashi-1 (blood and LP: 0 vs scattered) positive cells were detected in blood and lamina propria of UC samples as compared to controls. HGFR/CDX2 (blood: 0 vs 1 ± 0.59; LP: 0.8 ± 0.69 vs 2.06 ± 0.72, P < 0.05) and Musashi-1/CDX2 (blood and LP: 0 vs scattered) co-expressions were found in blood and lamina propria of UC samples. HGFR/CD133 and CD133/CDX2 co-expressions appeared only in UC lamina propria samples. CDX2, Lgr5 and Musashi-1 expressions in UC blood samples were not accompanied by CK20 mRNA expression.
  
  
  CONCLUSIONS
  In active UC, a portion of circulating HGFR-expressing cells are committed to the epithelial lineage, and may participate in mucosal regeneration by undergoing mesenchymal-to-epithelial transition.
authors.affiliation: Ferenc Sipos, Miklós Constantinovits, Györgyi Műzes, 2 Department of Internal Medicine, Semmelweis University, 1088 Budapest, Hungary.
authors.forename: Ferenc
authors.initials: F
authors.lastname: Sipos
authors.affiliation: Ferenc Sipos, Miklós Constantinovits, Györgyi Műzes, 2 Department of Internal Medicine, Semmelweis University, 1088 Budapest, Hungary.
authors.forename: Miklós
authors.initials: M
authors.lastname: Constantinovits
authors.affiliation: Ferenc Sipos, Miklós Constantinovits, Györgyi Műzes, 2 Department of Internal Medicine, Semmelweis University, 1088 Budapest, Hungary.
authors.forename: Gábor
authors.initials: G
authors.lastname: Valcz
authors.affiliation: Ferenc Sipos, Miklós Constantinovits, Györgyi Műzes, 2 Department of Internal Medicine, Semmelweis University, 1088 Budapest, Hungary.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Tulassay
authors.affiliation: Ferenc Sipos, Miklós Constantinovits, Györgyi Műzes, 2 Department of Internal Medicine, Semmelweis University, 1088 Budapest, Hungary.
authors.forename: Györgyi
authors.initials: G
authors.lastname: Műzes
mesh_terms: D000071916:AC133 Antigen; D015703:Antigens, CD / Q000032:analysis; D000071616:CDX2 Transcription Factor; D016022:Case-Control Studies; D002454:Cell Differentiation; D019070:Cell Lineage; D003093:Colitis, Ulcerative / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D003106:Colon / Q000737:chemistry* / Q000473:pathology / Q000503:physiopathology; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000032:analysis; D018398:Homeodomain Proteins / Q000032:analysis* / Q000097:blood / Q000235:genetics; D006801:Humans; D007413:Intestinal Mucosa / Q000737:chemistry* / Q000473:pathology / Q000503:physiopathology; D061346:Laser Capture Microdissection; D009419:Nerve Tissue Proteins / Q000032:analysis; D010455:Peptides / Q000032:analysis; D010641:Phenotype; D019859:Proto-Oncogene Proteins c-met / Q000032:analysis* / Q000097:blood / Q000235:genetics; D016601:RNA-Binding Proteins / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction; D012038:Regeneration; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013234:Stem Cells / Q000737:chemistry* / Q000473:pathology
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D000071616:CDX2 Transcription Factor; C491270:CDX2 protein, human; D006023:Glycoproteins; D018398:Homeodomain Proteins; C491297:MSI1 protein, human; D009419:Nerve Tissue Proteins; C000606023:PROM1 protein, human; D010455:Peptides; D016601:RNA-Binding Proteins; C491743:MET protein, human; D019859:Proto-Oncogene Proteins c-met
keywords: CD133; Caudal type homeobox 2; Hepatocyte-derived growth factor receptor; Leucine-rich repeat-containing G-protein coupled receptor 5; Musashi-1; Regeneration; Ulcerative colitis