TL1A Promotes Fibrogenesis in Colonic Fibroblasts via the TGF-β1/Smad3 Signaling Pathway

Abstract

Objective

Intestinal fibrosis is a refractory complication of inflammatory bowel disease (IBD). Tumor necrosis factor ligand-related molecule-1A (TL1A) is important for IBD-related intestinal fibrosis in a dextran sodium sulfate (DSS)-induced experimental colitis model. This study aimed to explore the effects of TL1A on human colonic fibroblasts.

Methods

A trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis model of LCK-CD2-TL1A-GFP transgenic (Tg) or wild-type (WT) mice was established to determine the effect and mechanism of TL1A on intestinal fibrosis. The human colonic fibroblast CCD-18Co cell line was treated concurrently with TL1A and human peripheral blood mononuclear cell (PBMC) supernatant. The proliferation and activation of CCD-18Co cells were detected by BrdU assays, flow cytometry, immunocytochemistry and Western blotting. Collagen metabolism was tested by Western blotting and real-time quantitative polymerase chain reaction (RT-qPCR).

Results

The level of collagen metabolism in the TNBS+ethyl alcohol (EtOH)/Tg group was greater than that in the TNBS+EtOH/WT group. Transforming growth factor-β1 (TGF-β1) and p-Smad3 in the TNBS+EtOH/Tg group were upregulated as compared with those in the TNBS+EtOH/WT group. The proliferation of CCD-18Co cells was promoted by the addition of human PBMC supernatant supplemented with 20 ng/mL TL1A, and the addition of human PBMC supernatant and TL1A increased CCD-18Co proliferation by 24.4% at 24 h. TL1A promoted cell activation and increased the levels of COL1A2, COL3A1, and TIMP-1 in CCD-18Co cells. Treatment of CCD-18Co cells with TL1A increased the expression of TGF-β1 and p-Smad3.

Conclusion

TL1A promotes TGF-β1-mediated intestinal fibroblast activation, proliferation, and collagen deposition and is likely related to an increase in the TGF-β1/Smad3 signaling pathway.

References

[1] YunSM, KimSH, KimEH. The Molecular Mechanism of Transforming Growth Factor-beta Signaling for Intestinal Fibrosis: A Mini-Review. Front Pharmacol, 2019, 10: 162

[2] D’HaensG, RiederF, FeaganBG, et al.. Challenges in the Pathophysiology, Diagnosis, and Management of Intestinal Fibrosis in Inflammatory Bowel Disease. Gastroenterology, 2022, 162(1): 26-31

[3] RiederF, FiocchiC. Intestinal fibrosis in IBD—a dynamic, multifactorial process. Nat Rev Gastroenterol Hepatol, 2009, 6(4): 228-235

[4] LiC, KuemmerleJF. The fate of myofibroblasts during the development of fibrosis in Crohn’s disease. J Dig Dis, 2020, 21(6): 326-331

[5] XuWD, LiR, HuangAF. Role of TL1A in Inflammatory Autoimmune Diseases: A Comprehensive Review. Front Immunol, 2022, 13: 891328

[6] HiranoA, YamazakiK, UmenoJ, et al.. Association study of 71 European Crohn’s disease susceptibility loci in a Japanese population. Inflamm Bowel Dis, 2013, 19: 526-533

[7] BarrettR, ZhangXL, KoonHW, et al.. Constitutive TL1A expression under colitogenic conditions modulates the severity and location of gut mucosal inflammation and induces fibrostenosis. Am J Pathol, 2012, 180: 636-649

[8] ZhengL, ZhangX, ChenJ, et al.. Sustained Tl1a (Tnfsf15) Expression on Both Lymphoid and Myeloid Cells Leads to Mild Spontaneous Intestinal Inflammation and Fibrosis. Eur J Microbiol Immunol (Bp), 2013, 3(1): 11-20

[9] JiaWX, YangMY, HanF, et al.. Effect and Mechanism of TL1A Expression on Epithelial Mesenchymal Transition during Chronic Colitis-Related Intestinal Fibrosis. Mediators Inflamm, 2021, 2021: 5927064

[10] LiH, SongJ, NiuGC, et al.. TL1A blocking ameliorates intestinal fibrosis in the T-cell transfer model of chronic colitis in mice. Pathol Res Pract, 2018, 214(2): 217-227

[11] AndohA, NishidaA. Molecular Basis of Intestinal Fibrosis in Inflammatory Bowel Disease. Inflamm Intest Dis, 2022, 7(3–4): 119-127

[12] TwohigJP, MarsdenM, CuffSM, et al.. The death receptor 3/TL1A pathway is essential for efficient development of antiviral CD4(+) and CD8(+) T-cell immunity. FASEB J, 2012, 26(8): 3575-3586

[13] WangJ, LinS, BrownJM, et al.. Novel mechanisms and clinical trial endpoints in intestinal fibrosis. Immunol Rev, 2021, 302(1): 211-227

[14] JonesGW, StumhoferJS, FosterT, et al.. Naive and activated T cells display differential responsiveness to TL1A that affects Th17 generation, maintenance, and proliferation. FASEB J, 2011, 25(1): 409-419

[15] SaikiaP, CrabbJS, DibbinLL, et al.. Quantitative proteomic comparison of myofibroblasts derived from bone marrow and cornea. Sci Rep, 2020, 10(1): 16717

[16] ChengRH, WangYP, ChangJY, et al.. Genetic Susceptibility and Protein Expression of Extracellular Matrix Turnover-Related Genes in Oral Submucous Fibrosis. Int J Mol Sci, 2020, 21(21): 8104

[17] DuY, MaJ, FanY, et al.. Naringenin: A Promising Therapeutic Agent against Organ Fibrosis. Oxid Med Cell Longev, 2021, 2021: 1210675

[18] WangR, WangD, WangH, et al.. Therapeutic Targeting of Nrf2 Signaling by Maggot Extracts Ameliorates Inflammation-Associated Intestinal Fibrosis in Chronic DSS-Induced Colitis. Front Immunol, 2021, 12: 670159

[19] MengXM, Nikolic-PatersonDJ, LanHY. TGF-beta: the master regulator of fibrosis. Nat Rev Nephrol, 2016, 12(6): 325-338

[20] ZhangH, ZhanY, ZhangY, et al.. Dual roles of TGF-beta signaling in the regulation of dental epithelial cell proliferation. J Mol Histol, 2021, 52(1): 77-86