Interleukin-13 Damages Intestinal Mucosa via TWEAK and Fn14 in Mice—A Pathway Associated With Ulcerative Colitis

doi:10.1053/j.gastro.2011.08.040

Gastroenterology

Volume 141, Issue 6, December 2011, Pages 2119-2129.e8

https://doi.org/10.1053/j.gastro.2011.08.040 Get rights and content

Background & Aims

TWEAK, a member of the tumor necrosis factor (TNF) superfamily, promotes intestinal epithelial cell injury and signals through the receptor Fn14 following irradiation-induced tissue damage and during development of colitis in mice. Interleukin (IL)-13, an effector of tissue damage in similar models, has been associated with the pathogenesis of ulcerative colitis (UC). We investigated interactions between TWEAK and IL-13 following mucosal damage in mice.

Methods

We compared patterns of gene expression in intestinal tissues from wild-type and TWEAK knockout mice following γ-irradiation. Intestinal explants from these mice were used to detect cell damage induced by IL-13 and TNF-α. Levels of messenger RNA for IL-13, TWEAK, and Fn14 were measured in mucosal samples from patients with UC.

Results

Based on gene expression analysis, TWEAK mediates γ-irradiation–induced epithelial cell cycle arrest and apoptosis. However, TWEAK alone did not induce damage or apoptosis of primary intestinal epithelial cells. On the other hand, exogenous IL-13 activated caspase-3 in naïve intestinal explants; this process required TWEAK, Fn14, and secretion of endogenous TNF-α which was mediated by ADAM17. Conversely, activation of caspase by exogenous TNF-α required IL-13, TWEAK, and Fn14. In mucosa from patients with UC, messenger RNA levels of IL-13, TWEAK, and Fn14 increased with level of disease severity.

Conclusions

IL-13–induced damage of intestinal epithelial cells requires TWEAK, its receptor (Fn14), and TNF-α. IL-13, TNF-α, TWEAK, and Fn14 could perpetuate and aggravate intestinal inflammation in patients with UC.

Section snippets

Mice

TWEAK¹⁶ or Fn14¹⁷ KO mice with BALB/c background were used for this study, except crossing IL-10 KO mice. IL-10 KO × TWEAK KO or IL-10 KO × Fn14 KO double KO mice were generated in our facility by crossing TWEAK KO mice or Fn14 KO mice with C57BL/6 background with IL-10 KO mice with C57BL/6 background obtained from The Jackson Laboratory (Bar Harbor, ME). All experimental protocols were approved by the local institutional animal care and use committees. For details, see Supplementary Materials

The TWEAK/Fn14 Pathway Is Necessary but Not Sufficient to Induce IEC Death

Recently we have shown that TWEAK/Fn14 promotes IEC death after 3-Gy irradiation in mice.¹⁵ Corresponding expression analysis showed that TWEAK messenger RNA (mRNA) was constitutively present in both the small and large intestine as expected. TWEAK mRNA tended to increase at 24 hours after γ-irradiation (Supplementary Figure 1A). Fn14 mRNA was also detected in naïve small and large intestine and significantly up-regulated 6 to 24 hours in jejunum and colon (Supplementary Figure 1A). We further

Discussion

In this study, we clearly showed a new linkage in that TWEAK/Fn14 is indispensable for IL-13–induced IEC damage. Importantly, our results indicate that IL-13 is able to trigger significant cell death using endogenous TWEAK/Fn14 and a latent form of TNF-α. In addition to establishing this novel link between IL-13 and the TWEAK/Fn14 pathway in IEC apoptosis, our data further elucidate that the proapoptotic mechanism of IL-13 in IECs is the promotion of TACE-mediated TNF-α shedding. The relevance

Acknowledgments

L.C.B. and T.D. share senior authorship.

The microarray files and information have been uploaded to the National Center for Biotechnology Information GEO public repository (accession number GSE25029).

References (33)

F. Heller et al.
Epithelial apoptosis is a prominent feature of the epithelial barrier disturbance in intestinal inflammation: effect of pro-inflammatory interleukin-13 on epithelial cell function
Mucosal Immunol
(2008)
R. Kawashima et al.
IL-13 receptor α2 promotes epithelial cell regeneration from radiation-induced small intestinal injury in mice
Gastroenterology
(2006)
F. Heller et al.
Interleukin-13 is the key effector Th2 cytokine in ulcerative colitis that affects epithelial tight junctions, apoptosis, and cell restitution
Gastroenterology
(2005)
I.J. Fuss et al.
The role of IL-13 and NK T cells in experimental and human ulcerative colitis
Mucosal Immunol
(2008)
H. Maecker et al.
TWEAK attenuates the transition from innate to adaptive immunity
Cell
(2005)
L.C. Burkly et al.
TWEAKing tissue remodeling by a multifunctional cytokine: role of TWEAK/Fn14 pathway in health and disease
Cytokine
(2007)
T. Saitoh et al.
TWEAK induces NF-kappaB2 p100 processing and long lasting NF-kappaB activation
J Biol Chem
(2003)
Y. Chicheportiche et al.
TWEAK, a new secreted ligand in the tumor necrosis factor family that weakly induces apoptosis
J Biol Chem
(1997)
A. Ikner et al.
TWEAK induces apoptosis through a death-signaling complex comprising receptor-interacting protein 1 (RIP1), Fas-associated death domain (FADD), and caspase-8
J Biol Chem
(2011)
D. Mandal et al.
REDOX regulation of IL-13 signaling in intestinal epithelial cells: usage of alternate pathways mediates distinct gene expression patterns
Cell Signal
(2010)

A. Roessner et al.

Oxidative stress in ulcerative colitis-associated carcinogenesis

Pathol Res Pract

(2008)

J.D. Schulzke et al.

Epithelial tight junctions in intestinal inflammation

Ann N Y Acad Sci

(2009)

I.J. Fuss et al.

Nonclassical CD1d-restricted NK T cells that produce IL-13 characterize an atypical Th2 response in ulcerative colitis

J Clin Invest

(2004)

W. Strober et al.

The signaling function of the IL-13Ralpha2 receptor in the development of gastrointestinal fibrosis and cancer surveillance

Curr Mol Med

(2009)

M.J. Kaplan et al.

The apoptotic ligands TRAIL, TWEAK, and Fas ligand mediate monocyte death induced by autologous lupus T cells

J Immunol

(2002)

S.A. Brown et al.

The Fn14 cytoplasmic tail binds tumour-necrosis-factor-receptor-associated factors 1, 2, 3 and 5 and mediates nuclear factor-kappaB activation

Biochem J

(2003)

Cited by (78)

Targeting uPA-uPAR interaction to improve intestinal epithelial barrier integrity in inflammatory bowel disease
2022, EBioMedicine
Citation Excerpt :
Moreover, crosstalk of these cell surface interactions is also important. For instance, TNF activates TNFR2 and induces redistribution of adhesion proteins to basolateral membranes of intestinal epithelial cells 11, while IL-13-induced damage of intestinal epithelial cells requires TWEAK and its receptor (Fn14).12 While a broad spectrum of IBD-omic studies look for disease relevant genes at the individual level, assessment of co-ordinately regulated ligand-receptor pairs is still lacking.
Loss of intestinal epithelial barrier integrity is a critical component of Inflammatory Bowel Disease (IBD) pathogenesis. Co-expression regulation of ligand-receptor pairs in IBD mucosa has not been systematically studied. Targeting ligand-receptor pairs which are induced in IBD mucosa and function in intestinal epithelial barrier integrity may provide novel therapeutics for IBD.
We performed transcriptomic meta-analysis on public IBD datasets combined with cell surface protein-protein-interaction (PPI) databases. We explored primary human/mouse intestinal organoids and Caco-2 cells for expression and function studies of uPA-uPAR (prime hits from the meta-analysis). Epithelial barrier integrity was measured by Trans-Epithelial Electrical Resistance (TEER), FITC-Dextran permeability and tight junction assessment. Genetic (CRISPR, siRNA and KO mice) and pharmacological (small molecules, neutralizing antibody and peptide inhibitors) approaches were applied. Mice deficient of uPAR were studied using the Dextran Sulfate Sodium (DSS)-induced colitis model.
The IBD ligand-receptor meta-analysis led to the discovery of a coordinated upregulation of uPA and uPAR in IBD mucosa. Both genes were significantly upregulated during epithelial barrier breakdown in primary intestinal organoids and decreased during barrier formation. Genetic inhibition of uPAR or uPA, or pharmacologically blocking uPA-uPAR interaction protects against cytokine-induced barrier breakdown. Deficiency of uPAR in epithelial cells leads to enhanced EGF/EGFR signalling, a known regulator of epithelial homeostasis and repair. Mice deficient of uPAR display improved intestinal barrier function in vitro and during DSS-induced colitis in vivo.
Our findings suggest that blocking uPA-uPAR interaction via pharmacological agents protects the epithelial barrier from inflammation-induced damage, indicating a potential therapeutic target for IBD.
The study was funded by Boehringer Ingelheim.
Patchouli oil ameliorates 5-fluorouracil-induced intestinal mucositis in rats via protecting intestinal barrier and regulating water transport
2020, Journal of Ethnopharmacology
Citation Excerpt :
Following 5-FU administration, the secretions of pro-inflammatory cytokines (TNF-α, IFN-γ, etc.) would increase, resulting in the infiltration of neutrophils and the promotion of the development of intestinal barrier injury. Besides, MPO, a marker of neutrophil infiltration, has been shown to be involved in the inflammatory process by causing oxidative reactions in inflammatory areas (Chami et al., 2018), and excessive expression of IL-13 has been shown to be involved in the inflammatory cascade, further enhancing gastrointestinal injury (Kawashima et al., 2011). Moreover, the alterations of inflammatory cytokines have been reported to be regulated by both NF-κB p65 and p38 MAPK activation (Huang et al., 2018; Justino et al., 2014).
Pogostemon cablin, commonly named “Guang-Huo-Xiang” in China, has long been renowned for its ability to dispel dampness and regulate gastrointestinal functions. Patchouli oil (P.oil), the major active fraction of Pogostemon cablin, has been traditionally used as the principal component of Chinese medicinal formulae to treat exterior syndrome and diarrhea. However, the effects of P.oil in treating 5-fluorouracil (5-FU)-induced intestinal mucositis have not yet been reported.
To investigate the protective effects of P.oil against 5-FU-induced intestinal mucositis and the mechanisms underlying these effects.
Sprague-Dawley rats were intraperitoneally injected with 5-FU (30 mg/kg) to establish an intestinal mucositis model. Meanwhile, rats with intestinal mucositis were orally administered with P.oil (25, 50, and 100 mg/kg). Histological analysis, ELISA (for detecting inflammatory cytokines and aquaporins), immunohistochemistry analysis (for examining caspases), qRT-PCR analysis (for assessment tight junctions), and western blotting analysis (for the assessment of TLR2/TLR4-MyD88 and VIP-cAMP-PKA signaling pathway-related proteins) were performed to estimate the protective effects of P.oil against intestinal mucositis and the mechanisms underlying these effects.
The histopathological assessment preliminarily exhibited that P.oil alleviated the 5-FU-induced damage to the intestinal structure. After P.oil administration, the elevation of the expression of cytokines (TNF-α, IFN-γ, and IL-13) decreased markedly and the activation of NF-κB and MAPK signaling was significantly inhibited. P.oil also increased the mRNA expression of ZO-1 and Occludin, thereby stabilizing intestinal barrier. In addition, P.oil decreased the expressions of caspase-8, caspase-3, and Bax, and increased the expression of Bcl-2, thereby reducing the apoptosis of the intestinal mucosa. These results were closely related to the regulation of the TLR2/TLR4-MyD88 signaling pathway. It has been indicated that P.oil possibly protected the intestinal barrier by reducing inflammation and apoptosis. Furthermore, this study showed that P.oil inhibited the abnormal expression of AQP3, AQP7, and AQP11 by regulating the VIP-cAMP-PKA signaling pathway. Furthermore, it restored the intestinal water absorption, thereby alleviating diarrhea.
P.oil ameliorated 5-FU-induced intestinal mucositis in rats via protecting intestinal barrier and regulating water transport.
FN14 Signaling Plays a Pathogenic Role in a Mouse Model of Experimental Autoimmune Myocarditis
2019, Journal of Cardiac Failure
Citation Excerpt :
Furthermore, it has been previously shown that cardiac fibroblasts as well as cardiomyocytes themselves can secrete proinflammatory cytokines.40 In addition, a connection between FN14 activation and increased cytokine expression could already be observed in other tissue injury models.11,28,30 Thus, by an increased FN14 expression, injured cardiac fibroblasts as well as cardiomyocytes seem to maintain an initial myocardial inflammation via activation of downstream signaling cascades resulting in proinflammatory cytokine expression.
The pathogenesis of inflammatory cardiomyopathy is affected by the activation of autoimmune-mediated cascades. To study these cascades, we developed an experimental model of troponin I (TnI)-induced autoimmune myocarditis (EAM). One factor playing a pivotal role in the context of autoimmune disorders is the receptor fibroblast growth factor-inducible 14 (FN14). Thus, the impact of FN14 in the development of autoimmune myocarditis was investigated.
TnI-immunization led to a significantly increased myocardial FN14 mRNA and protein expression in wild-type (wt) mice. To investigate the precise role of FN14 in EAM, FN14 knockout (ko) and wt littermates were immunized with TnI or control buffer. The animals were evaluated for cardiac parameters and indicators of myocardial injury. FN14 deficiency resulted in better cardiac performance, less myocardial inflammation, fibrosis, and cardiac damage. A lower myocardial mRNA expression of inflammatory cytokines and chemokines as well as their receptors could be demonstrated in TnI-immunized FN14ko compared to wt mice also immunized with TnI. Western blot analysis revealed a contribution of nuclear factor kappa-light-chain-enhancer of activated B cells to FN14-induced signaling cascades.
In the pathogenesis of autoimmune myocarditis, the inflammatory response to cardiac injury is attenuated in FN14ko mice. Thus, inhibition of FN14 in patients might represent a novel therapeutic strategy in the treatment of inflammatory cardiomyopathy.
Interleukin-13: A promising therapeutic target for autoimmune disease
2019, Cytokine and Growth Factor Reviews
Interleukin-13 (IL-13) was previously thought to be a redundant presence of IL-4, but in recent years its role in immunity, inflammation, fibrosis, and allergic diseases has become increasingly prominent. IL-13 can regulate several subtypes of T helper (Th) cells and affect their transformation, including Th1, Th2, T17, etc., thus it may play an important role in immune system. Previous studies have revealed that IL-13 is implicated in the pathogenesis of autoimmune diseases, such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), systemic sclerosis (SSc), ulcerative colitis (UC), type 1 diabetes (T1D), sjogren's syndrome (SS), etc. In this review, we will briefly discuss the biological features of IL-13 and summarize recent advances in the role of IL-13 in the development and pathogenesis of autoimmune diseases. This information may provide new perspectives and suggestions for the selection of therapeutic targets for autoimmune diseases.
TWEAK/Fn14 Is Overexpressed in Crohn's Disease and Mediates Experimental Ileitis by Regulating Critical Innate and Adaptive Immune Pathways
2019, Cellular and Molecular Gastroenterology and Hepatology
Crohn’s disease (CD) is a debilitating inflammatory disorder that affects more than 1.6 million people in North America alone. Members of the tumor necrosis factor superfamily are key regulators of intestinal inflammation; specifically, tumor necrosis factor–like weak inducer of apoptosis (TWEAK) and its receptor, fibroblast growth factor–inducible 14 (Fn14), are involved in normal and pathologic tissue remodeling. Our aim was to determine the role of TWEAK/Fn14 in CD and a murine model of CD-like ileitis (ie, SAMP1/YitFc [SAMP] strain).
SAMP mice deficient in Fn14 (SAMP × Fn14^-/-) were developed and a detailed time-course study was performed evaluating ileal tissues by histology and stereomicroscopy, as well as quantitative polymerase chain reaction and NanoString technology (Seattle, WA). Reciprocal bone marrow chimeras were generated to assess the relevance of Fn14 in hematopoietic vs nonhematopoietic compartments. Surgically resected intestinal tissues and mucosal biopsy specimens from patients with CD, ulcerative colitis, and healthy controls were analyzed for the expression of TWEAK/Fn14 by quantitative polymerase chain reaction, Western blot, immunohistochemistry, and immunofluorescence.
SAMP × Fn14^-/- showed a marked decrease in ileitis severity at 20 weeks of age compared with SAMP WT controls. Bone marrow chimeras showed that Fn14 was required in both hematopoietic and nonhematopoietic compartments for ileitis to develop. Transcriptome data showed multiple cellular pathways regulated by Fn14 signaling. Finally, increased expression of TWEAK and Fn14 was observed in tissue lesions from CD patients compared with ulcerative colitis and healthy controls.
TWEAK/Fn14 are up-regulated in CD, and also mediate experimental CD-like ileitis, by regulation of multiple innate and adaptive cellular pathways. Therefore, TWEAK/Fn14 may represent a novel therapeutic target for the treatment of small intestinal inflammation in CD.
Mucosal Expression of Type 2 and Type 17 Immune Response Genes Distinguishes Ulcerative Colitis From Colon-Only Crohn's Disease in Treatment-Naive Pediatric Patients
2017, Gastroenterology
There is controversy regarding the role of the type 2 immune response in the pathogenesis of ulcerative colitis (UC)—few data are available from treatment-naive patients. We investigated whether genes associated with a type 2 immune response in the intestinal mucosa are up-regulated in treatment-naive pediatric patients with UC compared with patients with Crohn’s disease (CD)-associated colitis or without inflammatory bowel disease (IBD), and whether expression levels are associated with clinical outcomes.
We used a real-time reverse-transcription quantitative polymerase chain reaction array to analyze messenger RNA (mRNA) expression patterns in rectal mucosal samples from 138 treatment-naive pediatric patients with IBD and macroscopic rectal disease, as well as those from 49 children without IBD (controls), enrolled in a multicenter prospective observational study from 2008 to 2012. Results were validated in real-time reverse-transcription quantitative polymerase chain reaction analyses of rectal RNA from an independent cohort of 34 pediatric patients with IBD and macroscopic rectal disease and 17 controls from Cincinnati Children’s Hospital Medical Center.
We measured significant increases in mRNAs associated with a type 2 immune response (interleukin [IL]5 gene, IL13, and IL13RA2) and a type 17 immune response (IL17A and IL23) in mucosal samples from patients with UC compared with patients with colon-only CD. In a regression model, increased expression of IL5 and IL17A mRNAs distinguished patients with UC from patients with colon-only CD (P = .001; area under the receiver operating characteristic curve, 0.72). We identified a gene expression pattern in rectal tissues of patients with UC, characterized by detection of IL13 mRNA, that predicted clinical response to therapy after 6 months (odds ratio [OR], 6.469; 95% confidence interval [CI], 1.553–26.94), clinical response after 12 months (OR, 6.125; 95% CI, 1.330–28.22), and remission after 12 months (OR, 5.333; 95% CI, 1.132–25.12).
In an analysis of rectal tissues from treatment-naive pediatric patients with IBD, we observed activation of a type 2 immune response during the early course of UC. We were able to distinguish patients with UC from those with colon-only CD based on increased mucosal expression of genes that mediate type 2 and type 17 immune responses. Increased expression at diagnosis of genes that mediate a type 2 immune response is associated with response to therapy and remission in pediatric patients with UC.

View all citing articles on Scopus

: Conflicts of interest The authors disclose no conflicts.

: Funding Supported in part by grants and contracts from the program Grants-in-Aid for Scientific Research (B) and Grants-in Aid for Young Scientists (B) from the Ministry of Education, Cultures, Sports, Science, and Technology; the Grant of National Center for Global Health and Medicine (21-110, 22-205), the Ministry of Health, Labor, and Welfare; Japan Health Sciences Foundation and Organization; and Health and Labor Sciences Research Grants for research on intractable diseases from the Ministry of Health, Labor and Welfare of Japan.

View full text

Original ResearchBasic and Translational—Alimentary TractInterleukin-13 Damages Intestinal Mucosa via TWEAK and Fn14 in Mice—A Pathway Associated With Ulcerative Colitis

Background & Aims

Methods

Results

Conclusions

Section snippets

Mice

The TWEAK/Fn14 Pathway Is Necessary but Not Sufficient to Induce IEC Death

Discussion

Acknowledgments

Mucosal Immunol

Gastroenterology

Gastroenterology

Mucosal Immunol

Cell

Cytokine

J Biol Chem

J Biol Chem

J Biol Chem

Cell Signal

Pathol Res Pract

Epithelial tight junctions in intestinal inflammation

Ann N Y Acad Sci

Nonclassical CD1d-restricted NK T cells that produce IL-13 characterize an atypical Th2 response in ulcerative colitis

J Clin Invest

The signaling function of the IL-13Ralpha2 receptor in the development of gastrointestinal fibrosis and cancer surveillance

Curr Mol Med

The apoptotic ligands TRAIL, TWEAK, and Fas ligand mediate monocyte death induced by autologous lupus T cells

J Immunol

The Fn14 cytoplasmic tail binds tumour-necrosis-factor-receptor-associated factors 1, 2, 3 and 5 and mediates nuclear factor-kappaB activation

Biochem J

Original Research
Basic and Translational—Alimentary Tract
Interleukin-13 Damages Intestinal Mucosa via TWEAK and Fn14 in Mice—A Pathway Associated With Ulcerative Colitis