Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 28, 2015; 21(28): 8569-8579
Published online Jul 28, 2015. doi: 10.3748/wjg.v21.i28.8569
Association of hepatocyte-derived growth factor receptor/caudal type homeobox 2 co-expression with mucosal regeneration in active ulcerative colitis
Ferenc Sipos, Miklós Constantinovits, Gábor Valcz, Zsolt Tulassay, Györgyi Műzes
Ferenc Sipos, Miklós Constantinovits, Györgyi Műzes, 2nd Department of Internal Medicine, Semmelweis University, 1088 Budapest, Hungary
Gábor Valcz, Zsolt Tulassay, Molecular Medicine Research Group, Hungarian Academy of Sciences, 1088 Budapest, Hungary
Author contributions: Sipos F, Constantinovits M, Valcz G, Tulassay Z and Műzes G substantially contributed to the conception and design of the study, acquisition, analysis and interpretation of data; all authors drafted the article and made critical revisions related to the intellectual content of the manuscript, and approved the final version of the article to be published.
Institutional review board statement: All routine colonic biopsy specimens and blood samples from the patients were taken after informed consent and ethical permission was obtained for participation in the study.
Conflict-of-interest statement: To the best of our knowledge, no conflict of interest exists.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Ferens Sipos, MD, PhD, Assistant Professor of Gastroenterology, Immunology and Internal Medicine, 2nd Department of Internal Medicine, Semmelweis University, Szentkirályi street 46, 1088 Budapest, Hungary. dr.siposf@gmail.com
Telephone: +36-1-2660926 Fax: +36-1-2660816
Received: December 2, 2014
Peer-review started: December 3, 2014
First decision: April 13, 2015
Revised: April 25, 2015
Accepted: June 15, 2015
Article in press: June 16, 2015
Published online: July 28, 2015
Processing time: 240 Days and 9.1 Hours
Abstract

AIM: 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.

CONCLUSION: 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.

Keywords: Hepatocyte-derived growth factor receptor; Caudal type homeobox 2; CD133; Musashi-1; Leucine-rich repeat-containing G-protein coupled receptor 5; Ulcerative colitis; Regeneration

Core tip: HGFR+ cells in the circulation and lamina propria of active ulcerative colitis (UC) could also co-express caudal type homeobox 2 (CDX2), an epithelial stem cell marker, thus suggesting that HGFR+ cells have committed to the epithelial lineage. The presence of CD133/CDX2 and Musashi-1/CDX2 double positive cells in the subepithelial layer supports that mesenchymal-to-epithelial transition might be a crucial event in tissue regeneration of active UC.