¹Antolić M., ¹Ognjenović A., ²Aralica G., ²Banić M., …...stem cell niche and...
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Crohn disease is a chronic inflammatory disease that can discontinu- ously affect different parts of the human digestive tract, but the most common are small intestine and colon. The incidence of Crohn disease is increasing annually in Western world countries (1). Crohn disease is characterized by transmural chronic inflammation including the forma- tion of granulomas, serositis and irregularity of crypt architecture. Pseudopyloric metaplasia is a very common histology feature in Crohn disease (2). The Wnt signaling pathway, whose part is Wnt-3α, has an important part in intestinal epithelial stem cell proliferation (3). In- creased expression of OLFM4 protein, marker of stem cell niche and Wnt-3α target gene, has been reported in the mucosal part of the in- testine of patients with active Crohn disease (4,5). The aim of the study was to explore changes in Wnt-3α distribution and ex- pression of OLFM4 protein in one of the Crohn disease features, pseudopy- loric metaplasia. Wnt-3α is part of the Wnt signaling pathway that is in- volved in the regulation of OLFM4 expression in the small intestine. OLFM4 plays a role in anti-inflammatory, apoptotic and proliferative processes in diseases of the digestive system (6). Pseudopyloric metaplasia is characteristic of intestinal ulcers in Crohn dis- ease, where it represents nonspecific reaction (7). The main pattern of pseudopyloric metaplasia in Crohn disease is the appearance of gastric glands/mucin-secreting cells characteristic for gastric pylorus, in small in- testine mucosa (8). Pseudopyloric metaplasia reflects the chronicity of in- flammatory processes in intestine mucosa where dilated intestine glands transit into aberrant gland form (9). One micron control and affected small intestine sections were stained using basic H&E, immunofluorescence (WNT3A antibody, Biorbyt, orb49054) and immunohistochemistry method (Anti-OLFM4 antibody, Abcam, ab85046). A study was performed on formalin-fixed paraf- fin-embedded intestinal tissues, obtained by resection either from Crohn patients (n=8) or carcinoma patients (controls). Out of these, three patients had pseudopyloric metaplasia. Histology slides were scanned on Zeiss Axioscan Z1 scanner (Zeiss, Jena, Germany). In the non-IBD small intestine, Wnt-3α is secreted by at least two types of intestinal epithelial secretory cells; Paneth cells situated in OLFM4 positive stem cell niche and enteroendocrine cells. In fulminant Crohn disease, elon- gated crypts showed an increased number of Wnt-3α positive Paneth cells within crypt stem cell niche and in ectopic position. OLFM4 labeled enlarged stem cell crypt zone. Most of the cells in pseudopyloric gland metaplasia were strongly Wnt-3α positive, while a small number of single cells were OLFM4 positive. In this study, we have shown that Wnt-3α secreting cells and OLFM4 positive cells are co-localized within intestine stem cell niche in control small intestine tissue. In Crohn disease Wnt-3α positive Paneth cells were found within elongated OLFM4 positive crypt stem cell niche and in ectopic position. Within pseudopyloric metaplastic glands Wnt-3α was strongly positive, while there was only small number of OLFM4 expressing epithelial cells. Hovde Ø., Moum B.A. (2012): Epidemiology and clinical course of Crohn's disease: results from observational studies. World J Gastroenterol, 18(15): 1723-1731 Magro F., Langner C., Driessen A., Ensari A., Geboes K., Mantzaris G.J., Villanacci V., Becheanu G., Borralho Nunes P., Cathomas G., Fries W., Jouret-Mourin A., Mescoli C., de Petris G., Rubio C.A., Shepherd N.A., Vieth M., Eliakim R.(2013): European consensus on the histopathology of inflammatory bowel disease. J Crohns Colitis, 7 (10): 827-51 Armbruster N. S., Stange E. F., Wehkamp J. (2017): In the Wnt of Paneth Cells: Immune-Epithelial Crosstalk in Small Intestinal Crohn’s Disease, Frontiers in Immunology, 8: 1204 Zou W.Y., Blutt S.E., Zeng X.L., Chen M.S., Lo Y.H. , Castillo-Azofeifa D., Klein O.D., Shroyer N.F., Donowitz M., Estes M. K.: Epithelial WNT Ligands Are Essential Drivers of Intestinal Stem Cell Activation. Cell Rep. 2018 23;22(4):1003-1015 Gersemann M., Becker S., Nuding S., Antoni L., Ott G., Fritz P., Oue N., Yasui W., Wehkamp J., Stange E. F.: Journal of Crohn's and Colitis (2012): Olfactomedin-4 is a glycoprotein secreted into mucus in active IBD, Journal of Crohn's and Colitis; 6, 425–434 Wang Xin-Yu, Chen Sheng-Hui, Zhang Ya-Nan, Xu Cheng-Fu (2018): Olfactomedin-4 in digestive diseases: A mini-review, World Journal of Gastroenterology, 24 (17): 1881-1887 Koukoulis, George K., Ke, Yong, Henley, John D.: Detection of Pyloric Metaplasia May Improve the Biopsy Diagnosis of Crohn's Ileitis. Journal of Clinical Gastroenterology. 34(2):141-143, February 2002. Ramai D., Changela K., Reddy M. (2017): Pyloric Gland Metaplasia of the Ileocecal Valve: Clinicopathologic Correlates of Inflammatory Bowel Disease, Cureus. 2017 Nov 3;9(11) Yokoyama I., Kozuka S., Ito K., Kubota K., Yokoyama Y. (1977): Gastric gland metaplasia in the small and large intestine, Gut, 18, 214-218 Study on formalin-fixed paraffin-embedded resected intestinal tissue from Crohn patients was approved by Hospital Ethical Committee. 1) 2) 3) 4) 5) 6) 7) 8) 9) Figure 1.: H&E stained tissue of control human small intestine. Figure 4.: H&E stained tissue of Crohn disease in human small intestine. Diffuse inflammation that consists of mononuclears, plasma cells and eosinophils in lamina propria and pseudopyloric metaplasia. Figure 2.: OLFM4 positive cells in stem cell niche in control human small intestine. Figure 5.: Strongly OLFM4 positive cells in stem cell niche of small intestine in patient with Crohn disease. Figure 3.: Wnt-3α positive Paneth and enteroendocrine cells in small intestine of control human small intestine. Figure 6.: Increased number of Wnt-3α positive Paneth cells in crypt stem cell niche in small intestine of patient with Crohn disease and Wnt-3α positive pseudopyloric metaplasia. Figure 7.: OLFM4 expression by elongated crypts and metaplastic epithelium in small intestine of patient with Crohn disease. Figure 8.: Wnt-3α expression by metaplastic epithelium in small intestine of patient with Crohn disease. ¹Antolić M., ¹Ognjenović A., ²Aralica G., ²Banić M., ¹Eraković Haber V., ¹Glojnarić I. and ¹Čužić S. ¹Fidelta Ltd., Prilaz baruna Filipovića 29, 10 000 Zagreb, Croatia ²Clinical Hospital Dubrava, Avenija Gojka Šuška 6, 10 000 Zagreb, Croatia info: [email protected]
Transcript of ¹Antolić M., ¹Ognjenović A., ²Aralica G., ²Banić M., …...stem cell niche and...
Crohn disease is a chronic inflammatory disease that can discontinu-ously affect different parts of the human digestive tract, but the most common are small intestine and colon. The incidence of Crohn disease is increasing annually in Western world countries (1). Crohn disease is characterized by transmural chronic inflammation including the forma-tion of granulomas, serositis and irregularity of crypt architecture. Pseudopyloric metaplasia is a very common histology feature in Crohn disease (2). The Wnt signaling pathway, whose part is Wnt-3α, has an important part in intestinal epithelial stem cell proliferation (3). In-creased expression of OLFM4 protein, marker of stem cell niche and Wnt-3α target gene, has been reported in the mucosal part of the in-testine of patients with active Crohn disease (4,5).
The aim of the study was to explore changes in Wnt-3α distribution and ex-pression of OLFM4 protein in one of the Crohn disease features, pseudopy-loric metaplasia. Wnt-3α is part of the Wnt signaling pathway that is in-volved in the regulation of OLFM4 expression in the small intestine. OLFM4 plays a role in anti-inflammatory, apoptotic and proliferative processes in diseases of the digestive system (6).Pseudopyloric metaplasia is characteristic of intestinal ulcers in Crohn dis-ease, where it represents nonspecific reaction (7). The main pattern of pseudopyloric metaplasia in Crohn disease is the appearance of gastric glands/mucin-secreting cells characteristic for gastric pylorus, in small in-testine mucosa (8). Pseudopyloric metaplasia reflects the chronicity of in-flammatory processes in intestine mucosa where dilated intestine glands transit into aberrant gland form (9).
One micron control and affected small intestine sections were stained using basic H&E, immunofluorescence (WNT3A antibody, Biorbyt, orb49054) and immunohistochemistry method (Anti-OLFM4 antibody, Abcam, ab85046). A study was performed on formalin-fixed paraf-fin-embedded intestinal tissues, obtained by resection either from Crohn patients (n=8) or carcinoma patients (controls). Out of these, three patients had pseudopyloric metaplasia. Histology slides were scanned on Zeiss Axioscan Z1 scanner (Zeiss, Jena, Germany).
In the non-IBD small intestine, Wnt-3α is secreted by at least two types of intestinal epithelial secretory cells; Paneth cells situated in OLFM4 positive stem cell niche and enteroendocrine cells. In fulminant Crohn disease, elon-gated crypts showed an increased number of Wnt-3α positive Paneth cells within crypt stem cell niche and in ectopic position. OLFM4 labeled enlarged stem cell crypt zone. Most of the cells in pseudopyloric gland metaplasia were strongly Wnt-3α positive, while a small number of single cells were OLFM4 positive.
In this study, we have shown that Wnt-3α secreting cells and OLFM4 positive cells are co-localized within intestine stem cell niche in control small intestine tissue. In Crohn disease Wnt-3α positive Paneth cells were found within elongated OLFM4 positive crypt stem cell niche and in ectopic position. Within pseudopyloric metaplastic glands Wnt-3α was strongly positive, while there was only small number of OLFM4 expressing epithelial cells.
Hovde Ø., Moum B.A. (2012): Epidemiology and clinical course of Crohn's disease: results from observational studies. World J Gastroenterol, 18(15): 1723-1731Magro F., Langner C., Driessen A., Ensari A., Geboes K., Mantzaris G.J., Villanacci V., Becheanu G., Borralho Nunes P., Cathomas G., Fries W., Jouret-Mourin A., Mescoli C., de Petris G., Rubio C.A., Shepherd N.A., Vieth M., Eliakim R.(2013): European consensus on the histopathology of inflammatory bowel disease. J Crohns Colitis, 7 (10): 827-51Armbruster N. S., Stange E. F., Wehkamp J. (2017): In the Wnt of Paneth Cells: Immune-Epithelial Crosstalk in Small Intestinal Crohn’s Disease, Frontiers in Immunology, 8: 1204Zou W.Y., Blutt S.E., Zeng X.L., Chen M.S., Lo Y.H. , Castillo-Azofeifa D., Klein O.D., Shroyer N.F., Donowitz M., Estes M. K.: Epithelial WNT Ligands Are Essential Drivers of Intestinal Stem Cell Activation. Cell Rep. 2018 23;22(4):1003-1015Gersemann M., Becker S., Nuding S., Antoni L., Ott G., Fritz P., Oue N., Yasui W., Wehkamp J., Stange E. F.: Journal of Crohn's and Colitis (2012): Olfactomedin-4 is a glycoproteinsecreted into mucus in active IBD, Journal of Crohn's and Colitis; 6, 425–434Wang Xin-Yu, Chen Sheng-Hui, Zhang Ya-Nan, Xu Cheng-Fu (2018): Olfactomedin-4 in digestive diseases: A mini-review, World Journal of Gastroenterology, 24 (17): 1881-1887Koukoulis, George K., Ke, Yong, Henley, John D.: Detection of Pyloric Metaplasia May Improve the Biopsy Diagnosis of Crohn's Ileitis. Journal of Clinical Gastroenterology. 34(2):141-143, February 2002.Ramai D., Changela K., Reddy M. (2017): Pyloric Gland Metaplasia of the Ileocecal Valve: Clinicopathologic Correlates of Inflammatory Bowel Disease, Cureus. 2017 Nov 3;9(11)Yokoyama I., Kozuka S., Ito K., Kubota K., Yokoyama Y. (1977): Gastric gland metaplasia in the small and large intestine, Gut, 18, 214-218
Study on formalin-fixed paraffin-embeddedresected intestinal tissue from Crohn patients was approved by Hospital Ethical Committee. 1)
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3)4)
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6)7)
8)9)
Figure 1.: H&E stained tissue of control human smallintestine.
Figure 4.: H&E stained tissue of Crohn disease in human small intestine. Diffuse inflammation that consists of mononuclears, plasma cells and eosinophils in lamina
propria and pseudopyloric metaplasia.
Figure 2.: OLFM4 positive cells in stem cell niche in control human small intestine.
Figure 5.: Strongly OLFM4 positive cells in stem cell niche of small intestine in patient with Crohn disease.
Figure 3.: Wnt-3α positive Paneth and enteroendocrine cells in small intestine of control human small intestine.
Figure 6.: Increased number of Wnt-3α positive Paneth cells in crypt stem cell niche in small intestine of patient with Crohn disease and Wnt-3α positive
pseudopyloric metaplasia.
Figure 7.: OLFM4 expression by elongated crypts and metaplasticepithelium in small intestine of patient with Crohn disease.
Figure 8.: Wnt-3α expression by metaplastic epithelium in smallintestine of patient with Crohn disease.
¹Antolić M., ¹Ognjenović A., ²Aralica G., ²Banić M., ¹Eraković Haber V., ¹Glojnarić I. and ¹Čužić S.
¹Fidelta Ltd., Prilaz baruna Filipovića 29, 10 000 Zagreb, Croatia²Clinical Hospital Dubrava, Avenija Gojka Šuška 6, 10 000 Zagreb, Croatia
info: [email protected]
