Fibrogenesis in Human Mucosa and Muscularis Precision-Cut Intestinal Slices

Overview

Journal Cells

Publisher MDPI

Specialties Biochemistry
Biophysics
Cell Biology
Molecular Biology

Date 2024 Jul 12

PMID 38994938

Authors

Carin Biel

Anniek Kastermans

Janneke Heidema

Martin Pehrsson

Charlotte Henstra

Joachim Mortensen

Klaas Nico Faber

Peter Olinga

Affiliations

Soon will be listed here.

Abstract

In Crohn's Disease (CD), intestinal fibrosis is a prevalent yet unresolved complication arising from chronic and transmural inflammation. The histological assessment of CD intestines shows changes in tissue morphology in all the layers, including the mucosa and muscularis. This study aimed to determine the differences in fibrogenesis between mucosa and muscularis. Human precision-cut intestinal slices (hPCIS) were prepared from human intestine mucosa and muscularis and treated with TGF-β1 and/or PDGF-BB for 72 h. Gene and protein expression and matrix metalloproteinase (MMP) activity were determined. The basal gene expression of various fibrosis markers was higher in muscularis compared to mucosa hPCIS. During incubation, Pro-Collagen-1A1 secretion increased in muscularis but not in mucosa hPCIS. MMP gene expression increased during incubation in mucosa and muscularis hPCIS, except for MMP9, MMP12, and MMP13 in muscularis hPCIS. Incubation with TGF-β1 caused increased COL1A1 expression in the mucosa but not in muscularis hPCIS. In muscularis hPCIS, TGF-β1 treatment caused a decrease in MMP1 and CTSK expression, while MMP13 was increased. In the presence of TGF-β1, protease inhibitor expression was stable, except for SERPINE1, which was increased in muscularis hPCIS. We conclude that fibrogenesis is more pronounced in muscularis hPCIS compared to mucosa hPCIS, especially when stimulated with TGF-β1.

References

Hinz B, Gabbiani G . Cell-matrix and cell-cell contacts of myofibroblasts: role in connective tissue remodeling. Thromb Haemost. 2003; 90(6):993-1002. DOI: 10.1160/TH03-05-0328. View

Pucilowska J, McNaughton K, Mohapatra N, Hoyt E, Zimmermann E, Sartor R . IGF-I and procollagen alpha1(I) are coexpressed in a subset of mesenchymal cells in active Crohn's disease. Am J Physiol Gastrointest Liver Physiol. 2000; 279(6):G1307-22. DOI: 10.1152/ajpgi.2000.279.6.G1307. View

Kumagai S, Ohtani H, Nagai T, Funa K, Hiwatashi N, Shimosegawa . Platelet-derived growth factor and its receptors are expressed in areas of both active inflammation and active fibrosis in inflammatory bowel disease. Tohoku J Exp Med. 2002; 195(1):21-33. DOI: 10.1620/tjem.195.21. View

de Bruyn J, van den Brink G, Steenkamer J, Buskens C, Bemelman W, Meisner S . Fibrostenotic Phenotype of Myofibroblasts in Crohn's Disease is Dependent on Tissue Stiffness and Reversed by LOX Inhibition. J Crohns Colitis. 2018; 12(7):849-859. DOI: 10.1093/ecco-jcc/jjy036. View

Iswandana R, Irianti M, Oosterhuis D, Hofker H, Merema M, de Jager M . Regional Differences in Human Intestinal Drug Metabolism. Drug Metab Dispos. 2018; 46(12):1879-1885. DOI: 10.1124/dmd.118.083428. View

Pham B, van Haaften W, Oosterhuis D, Nieken J, de Graaf I, Olinga P . Precision-cut rat, mouse, and human intestinal slices as novel models for the early-onset of intestinal fibrosis. Physiol Rep. 2015; 3(4). PMC: 4425951. DOI: 10.14814/phy2.12323. View

Pozzer D, Invernizzi R, Blaauw B, Cantoni O, Zito E . Ascorbic Acid Route to the Endoplasmic Reticulum: Function and Role in Disease. Antioxid Redox Signal. 2019; 34(11):845-855. DOI: 10.1089/ars.2019.7912. View

Alfredsson J, Wick M . Mechanism of fibrosis and stricture formation in Crohn's disease. Scand J Immunol. 2020; 92(6):e12990. PMC: 7757243. DOI: 10.1111/sji.12990. View

van Haaften W, Blokzijl T, Hofker H, Olinga P, Dijkstra G, Bank R . Intestinal stenosis in Crohn's disease shows a generalized upregulation of genes involved in collagen metabolism and recognition that could serve as novel anti-fibrotic drug targets. Therap Adv Gastroenterol. 2020; 13:1756284820952578. PMC: 7457685. DOI: 10.1177/1756284820952578. View

10.

Leeming D, Larsen D, Zhang C, Hi Y, Veidal S, Nielsen R . Enzyme-linked immunosorbent serum assays (ELISAs) for rat and human N-terminal pro-peptide of collagen type I (PINP)--assessment of corresponding epitopes. Clin Biochem. 2010; 43(15):1249-56. DOI: 10.1016/j.clinbiochem.2010.07.025. View

11.

Chen W, Lu C, Hirota C, Iacucci M, Ghosh S, Gui X . Smooth Muscle Hyperplasia/Hypertrophy is the Most Prominent Histological Change in Crohn's Fibrostenosing Bowel Strictures: A Semiquantitative Analysis by Using a Novel Histological Grading Scheme. J Crohns Colitis. 2016; 11(1):92-104. DOI: 10.1093/ecco-jcc/jjw126. View

12.

Severi C, Sferra R, Scirocco A, Vetuschi A, Pallotta N, Pronio A . Contribution of intestinal smooth muscle to Crohn's disease fibrogenesis. Eur J Histochem. 2015; 58(4):2457. PMC: 4289851. DOI: 10.4081/ejh.2014.2457. View

13.

di Sabatino A, Jackson C, Pickard K, Buckley M, Rovedatti L, Leakey N . Transforming growth factor beta signalling and matrix metalloproteinases in the mucosa overlying Crohn's disease strictures. Gut. 2009; 58(6):777-89. DOI: 10.1136/gut.2008.149096. View

14.

Mao R, Doyon G, Gordon I, Li J, Lin S, Wang J . Activated intestinal muscle cells promote preadipocyte migration: a novel mechanism for creeping fat formation in Crohn's disease. Gut. 2021; 71(1):55-67. PMC: 8286985. DOI: 10.1136/gutjnl-2020-323719. View

15.

Van Haaften W, Mortensen J, Karsdal M, Bay-Jensen A, Dijkstra G, Olinga P . Misbalance in type III collagen formation/degradation as a novel serological biomarker for penetrating (Montreal B3) Crohn's disease. Aliment Pharmacol Ther. 2017; 46(1):26-39. PMC: 6221070. DOI: 10.1111/apt.14092. View

16.

Afratis N, Selman M, Pardo A, Sagi I . Emerging insights into the role of matrix metalloproteases as therapeutic targets in fibrosis. Matrix Biol. 2018; 68-69:167-179. DOI: 10.1016/j.matbio.2018.02.007. View

17.

di Mola F, Friess H, Scheuren A, Di Sebastiano P, Graber H, Egger B . Transforming growth factor-betas and their signaling receptors are coexpressed in Crohn's disease. Ann Surg. 1999; 229(1):67-75. PMC: 1191610. DOI: 10.1097/00000658-199901000-00009. View

18.

Li C, Kuemmerle J . The fate of myofibroblasts during the development of fibrosis in Crohn's disease. J Dig Dis. 2020; 21(6):326-331. DOI: 10.1111/1751-2980.12852. View

19.

Ortega M, Rios-Navarro C, Gavara J, de Dios E, Perez-Sole N, Marcos-Garces V . Meta-Analysis of Extracellular Matrix Dynamics after Myocardial Infarction Using RNA-Sequencing Transcriptomic Database. Int J Mol Sci. 2022; 23(24). PMC: 9779146. DOI: 10.3390/ijms232415615. View

20.

Bourgonje A, Alexdottir M, Otten A, Loveikyte R, Bay-Jensen A, Pehrsson M . Serological biomarkers of type I, III and IV collagen turnover are associated with the presence and future progression of stricturing and penetrating Crohn's disease. Aliment Pharmacol Ther. 2022; 56(4):675-693. PMC: 9544881. DOI: 10.1111/apt.17063. View