An INF-STAT Axis Augments Tissue Damage and Inflammation in a Mouse Model of Crohn's Disease

Blocking interferon-function by therapeutic intervention of the JAK-STAT-axis is a novel promising treatment option for inflammatory bowel disease (IBD). Although JAK inhibitors have proven efficacy in patients with active ulcerative colitis (UC), they failed to induce clinical remission in patients with Crohn's disease (CD). This finding strongly implicates a differential contribution of JAK signaling in both entities. Here, we dissected the contribution of different STAT members downstream of JAK to inflammation and barrier dysfunction in a mouse model of Crohn's disease like ileitis and colitis (Casp8ΔIEC mice). Deletion of STAT1 in Casp8ΔIEC mice was associated with reduced cell death and a partial rescue of Paneth cell function in the small intestine. Likewise, organoids derived from the small intestine of these mice were less sensitive to cell death triggered by IBD-key cytokines such as TNFα or IFNs. Further functional in vitro and in vivo analyses revealed the impairment of MLKL-mediated necrosis as a result of deficient STAT1 function, which was in turn associated with improved cell survival. However, a decrease in inflammatory cell death was still associated with mild inflammation in the small intestine. The impact of STAT1 signaling on gastrointestinal inflammation dependent on the localization of inflammation, as STAT1 is essential for intestinal epithelial cell death regulation in the small intestine, whereas it is not the key factor for intestinal epithelial cell death in the context of colitis. Of note, additional deletion of STAT2 was not sufficient to restore Paneth cell function but strongly ameliorated ileitis. In summary, we provide here compelling molecular evidence that STAT1 and STAT2, both contribute to intestinal homeostasis, but have non-redundant functions. Our results further demonstrate that STATs individually affect the distinct pathophysiology of inflammation in the ileum and colon, respectively, which might explain the diverse outcome of JAK inhibitors on inflammatory bowel diseases.

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Crohn's disease-derived monocytes fail to induce Paneth cell defensins

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1510084112 ◽

2015 ◽

Vol 112 (45) ◽

pp. 14000-14005 ◽

Cited By ~ 34

Author(s):

Lioba F. Courth ◽

Maureen J. Ostaff ◽

Daniela Mailänder-Sánchez ◽

Nisar P. Malek ◽

Eduard F. Stange ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Mononuclear Cells ◽

Cell Function ◽

Paneth Cell ◽

Constitutive Expression ◽

Paneth Cells ◽

Intestinal Epithelial ◽

Peripheral Blood Mononuclear ◽

Wnt Ligands

Crohn’s disease (CD) is associated with a multitude of genetic defects, many of which likely affect Paneth cell function. Paneth cells reside in the small intestine and produce antimicrobial peptides essential for the host barrier, principally human α-defensin 5 (HD5) and HD6. Patients with CD of the ileum are characterized by reduced constitutive expression of these peptides and, accordingly, compromised antimicrobial barrier function. Here, we present a previously unidentified regulatory mechanism of Paneth cell defensins. Using cultures of human ileal tissue, we showed that the secretome of peripheral blood mononuclear cells (PBMCs) from healthy controls restored the attenuated Paneth cell α-defensin expression characteristic of patients with ileal CD. Analysis of the Wnt pathway in both cultured biopsies and intestinal epithelial cells implicated Wnt ligands driving the PBMC effect, whereas various tested cytokines were ineffective. We further detected another defect in patients with ileal CD, because the PBMC secretomes derived from patients with CD were unable to restore the reduced HD5/HD6 expression. Accordingly, analysis of PBMC subtypes showed that monocytes of patients with CD express significantly lower levels of canonical Wnt ligands, including Wnt3, Wnt3a, Wnt1, and wntless Wnt ligand secretion mediator (Evi/Wls). These studies reveal an important cross-talk between bone marrow-derived cells and epithelial secretory Paneth cells. Defective Paneth cell-mediated innate immunity due to inadequate Wnt ligand stimulation by monocytes provides an additional mechanism in CD. Because defects of Paneth cell function stemming from various etiologies are overcome by Wnt ligands, this mechanism is a potential therapeutic target for this disease.

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Faecal Biomarkers in Inflammatory Bowel Diseases: Calprotectin Versus Lipocalin-2—a Comparative Study

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jjaa124 ◽

2020 ◽

Cited By ~ 3

Author(s):

Andreas Zollner ◽

Andreas Schmiderer ◽

Simon J Reider ◽

Georg Oberhuber ◽

Alexandra Pfister ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Inflammatory Bowel Diseases ◽

Low Grade ◽

Intestinal Epithelial ◽

Lipocalin 2 ◽

Monitoring Tools ◽

Bowel Diseases ◽

Inflammatory Bowel ◽

Histological Remission

Abstract Background and Aims Faecal biomarkers, particularly calprotectin [FCAL], have become important diagnostic and monitoring tools in inflammatory bowel diseases [IBD]. As FCAL is mainly produced by neutrophils, we hypothesised that faecal lipocalin-2 [FLCN2], also expressed by intestinal epithelial cells [IEC], could be beneficial in specific clinical situations. Methods We compared clinical and endoscopic activity-related correlations between FCAL and FLCN2, assayed from the same sample, in a cohort of 132 patients (72 Crohn’s disease [CD]) and 40 controls. A detailed analysis of cellular origins was done by confocal microscopy and flow cytometry. To evaluate the potential to detect low-grade inflammation, we studied faecal and tissue concentrations in a cohort with clinical, endoscopic, and histological remission. Results There was an excellent correlation between FCAL and FLCN2 [rS = 0.87, p <0.001] and comparable sensitivity and specificity to predict clinical and endoscopic disease activity, with optimal thresholds for endoscopic activity of 73.4 and 1.98 µg/g in ulcerative colitis [UC] and 78.4 and 0.56 µg/g in Crohn’s disease for FCAL and FLCN2, respectively. Strong co-expression of both proteins was observed in granulocytes and macrophages. IECs expressed LCN2 but not CAL. In our IBD cohort in deep remission neither FCAL nor FLCN2 was different from controls; yet mucosal LCN2 but not CAL expressions remained elevated in the rectum of UC and the ileum of CD patients. Conclusions This study corroborates the diagnostic equivalence of FLCN2 and FCAL in IBD. In remission, persistent mucosal overexpression renders LCN2 an attractive candidate for molecular inflammation warranting further investigation.

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Single-cell analysis of colonic epithelium reveals unexpected shifts in cellular composition and molecular phenotype in treatment-naïve adult Crohn’s disease

10.1101/2021.01.13.426602 ◽

2021 ◽

Author(s):

Matt Kanke ◽

Meaghan M. Kennedy ◽

Sean Connelly ◽

Matthew Schaner ◽

Michael T. Shanahan ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Single Cell ◽

Association Studies ◽

Paneth Cell ◽

Genome Wide Association Studies ◽

Intestinal Epithelial ◽

Cell Type ◽

Aberrant Expression ◽

Treatment Naïve

AbstractThe intestinal epithelial barrier is comprised of a monolayer of specialized intestinal epithelial cells (IECs) that are critical in maintaining gut mucosal homeostasis. Dysfunction within various IEC fractions can increase intestinal permeability, resulting in a chronic and debilitating condition known as Crohn’s disease (CD). Defining the molecular changes in each IEC type in CD will contribute to an improved understanding of the pathogenic processes and the identification of potential therapeutic targets. Here we performed, for the first time at single-cell resolution, a direct comparison of the colonic epithelial cellular and molecular landscape between treatment-naïve adult CD and non-IBD control patients. Our analysis revealed that in CD patients there is a significant skew in the colonic epithelial cellular distribution away from canonical LGR5+ stem cells, located at the crypt-bottom, and toward one specific subtype of mature colonocytes, located at the crypt-top. Further analysis revealed unique changes to gene expression programs in every major cell type, including a previously undescribed suppression in CD of most enteroendocrine driver genes as well as L-cell markers including GCG. We also dissect a previously poorly understood SPIB+ cell cluster, revealing at least four sub-clusters that exhibit unique features. One of these SPIB+ sub-clusters expresses crypt-top colonocyte markers and is significantly up-regulated in CD, whereas another sub-cluster strongly expresses and stains positive for lysozyme (albeit no other canonical Paneth cell marker), which surprisingly is greatly reduced in expression in CD. Finally, through integration with data from genome-wide association studies, we show that genes implicated in CD risk exhibit heretofore unknown cell-type specific patterns of aberrant expression in CD, providing unprecedented insight into the potential biological functions of these genes.

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Lipid and Bile Acid Dysmetabolism in Crohn’s Disease

Journal of Immunology Research ◽

10.1155/2018/7270486 ◽

2018 ◽

Vol 2018 ◽

pp. 1-6 ◽

Cited By ~ 10

Author(s):

Koji Uchiyama ◽

Hisashi Kishi ◽

Wataru Komatsu ◽

Masanori Nagao ◽

Shuji Ohhira ◽

...

Keyword(s):

Fatty Acids ◽

Small Intestine ◽

Crohn’S Disease ◽

Bile Acid ◽

Crohn's Disease ◽

Terminal Ileum ◽

Cell Function ◽

Loss Of Function ◽

Bile Acid Malabsorption

Crohn’s disease is one of the systemic autoimmune diseases. It commonly affects the small intestine and colon but may involve any portion of the gastrointestinal tract from the mouth to the anus. The most affected area by Crohn’s disease is the distal part of the small intestine, in which the bile acid molecules are most efficiently reabsorbed. Bile acids form mixed micelles together with fatty acids, which function as a transport vehicle to deliver fatty acids to the apical membrane of enterocytes for absorption. Therefore, if the terminal ileum is impaired, bile acid malabsorption may occur, which may cause congenital diarrhoea in Crohn’s disease. Similarly, the impairment of the terminal ileum also induces fatty acid malabsorption, which may influence the role of fatty acids in Crohn’s disease. In contrast, a recent study reported that multidrug resistance protein 1 (MDR1) regulated effector T-cell function in the ileum from bile acid-driven oxidative stress and MDR1 loss of function in a subset of patients with Crohn’s disease. However, the role of consumption of fatty acids in Crohn’s disease remains to be fully elucidated. This review is aimed at providing an overview of some recent developments in research of Crohn’s disease from comprehensive perspective with a focus on the connection between disease location and behaviour, lipid diets, and bile acid malabsorption.

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P061 VALUE OF US AND MRE IMAGES IN THE DIAGNOSTICS OF ACTIVE CROHN’S DISEASE IN CHILDREN

Inflammatory Bowel Diseases ◽

10.1093/ibd/zaa010.036 ◽

2020 ◽

Vol 26 (Supplement_1) ◽

pp. S15-S16

Author(s):

Monika Zbroja ◽

Weronika Cyranka ◽

Maryla Kuczynska ◽

Monika Piekarska ◽

Karolina Siejka ◽

...

Keyword(s):

Small Intestine ◽

Crohn’S Disease ◽

Crohn's Disease ◽

Active Phase ◽

Chronic Inflammatory Bowel Disease ◽

Inflammatory Infiltration ◽

Inflammatory Reactions ◽

Linear Probe ◽

The Us ◽

Inflammatory Bowel

Abstract Introduction Crohn’s disease is classified as chronic inflammatory bowel disease. The incidence of it in Europe ranges from 1 to almost 11.4 per 100,000 population per year. Ultrasound and magnetic resonance enterography examinations play an important role in imaging diagnostics of inflammatory bowel lesions. They allow for recognising and monitoring changes during therapy as well as assessing complications such as fistula or abscess. Materials and Methods 36 children were included in the study: 16 boys and 20 girls with an active phase of Crohn’s disease. Each patient underwent intestinal ultrasound examination with a high frequency 7–12 Mhz linear probe and MRE with intravenous administration of a contrast agent. Results All patients showed a significant correlation between ultrasound and MRE. In the US examination all children had thickened, low echogenic wall showing varying degrees of vascular flow signals. Additionally, in 8 patients Bauhin’ valve edema was visible. In 16 children, inflammatory infiltration of the periintestinal fat around the affected segment of the intestine was found. In all patients a mesenteric lymphadenopathy in the short axis of 10-15mm was visible. MRE confirmed the presence of the confirmed ideal lesions and Bauhin’ valve edema in all 36 children. In addition, in 4 patients small intestine fistulas were found whereas abscess was observed in another 4 patients. In contrast-enhanced images, a vivid enhancement of the affected bowel section was revealed and in 10 children inflammatory reactions of peri-intestinal fat was demonstrated. Conclusion US and MRE are reliable tools in diagnosis of enteric inflammatory disease of the small intestine, evaluation of disease activity and assessment of potential complications. They are complementary elements in diagnostics of Crohn’s disease.

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Interferon Lambda Promotes Paneth Cell Death Via STAT1 Signaling in Mice and Is Increased in Inflamed Ileal Tissues of Patients With Crohn’s Disease

Gastroenterology ◽

10.1053/j.gastro.2019.07.031 ◽

2019 ◽

Vol 157 (5) ◽

pp. 1310-1322.e13 ◽

Cited By ~ 14

Author(s):

Claudia Günther ◽

Barbara Ruder ◽

Iris Stolzer ◽

Heidrun Dorner ◽

Gui-Wei He ◽

...

Keyword(s):

Cell Death ◽

Crohn’S Disease ◽

Crohn's Disease ◽

Paneth Cell ◽

Interferon Lambda ◽

Stat1 Signaling

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Differential expression of TGF-β isoforms by normal and inflammatory bowel disease intestinal myofibroblasts

AJP Cell Physiology ◽

10.1152/ajpcell.00048.2001 ◽

2002 ◽

Vol 282 (1) ◽

pp. C172-C182 ◽

Cited By ~ 104

Author(s):

B. C. McKaig ◽

K. Hughes ◽

P. J. Tighe ◽

Y. R. Mahida

Keyword(s):

Ulcerative Colitis ◽

Crohn’S Disease ◽

Crohn's Disease ◽

Wound Repair ◽

Transforming Growth Factor ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Proliferative Capacity ◽

Inflammatory Bowel ◽

Lower Expression

First published September 5, 2001; 10.1152/ajpcell. 00048.2001.—Intestinal strictures are frequent in Crohn's disease but not ulcerative colitis. We investigated the expression of transforming growth factor (TGF)-β isoforms by isolated and cultured primary human intestinal myofibroblasts and the responsiveness of these cells and intestinal epithelial cells to TGF-β isoforms. Normal intestinal myofibroblasts released predominantly TGF-β3 and ulcerative colitis myofibroblasts expressed both TGF-β1 and TGF-β3, whereas in myofibroblast cultures from fibrotic Crohn's disease tissue, there was significantly lower expression of TGF-β3 but enhanced release of TGF-β2. These distinctive patterns of TGF-β isoform release were sustained through several myofibroblast passages. Proliferation of Crohn's disease myofibroblasts was significantly greater than that of myofibroblasts derived from normal and ulcerative colitis tissue. In contrast to cells from normal and ulcerative colitis tissue, neutralization of the three TGF-β isoforms did not affect the proliferation of Crohn's disease intestinal myofibroblasts. Studies on the effect of recombinant TGF-β isoforms on epithelial restitution and proliferation suggest that TGF-β2 may be the least effective of the three isoforms in intestinal wound repair. In conclusion, the enhanced release of TGF-β2 but reduced expression of TGF-β3 by Crohn's disease intestinal myofibroblasts, together with their enhanced proliferative capacity, may lead to the development of intestinal strictures.

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Defective ATG16L1-mediated removal of IRE1α drives Crohn’s disease–like ileitis

Journal of Experimental Medicine ◽

10.1084/jem.20160791 ◽

2017 ◽

Vol 214 (2) ◽

pp. 401-422 ◽

Cited By ~ 79

Author(s):

Markus Tschurtschenthaler ◽

Timon E. Adolph ◽

Jonathan W. Ashcroft ◽

Lukas Niederreiter ◽

Richa Bharti ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Er Stress ◽

Paneth Cell ◽

Protective Role ◽

Dependent Manner ◽

Intestinal Epithelial ◽

Unfolded Protein ◽

Antimicrobial Function ◽

The Unfolded Protein Response

ATG16L1T300A, a major risk polymorphism in Crohn’s disease (CD), causes impaired autophagy, but it has remained unclear how this predisposes to CD. In this study, we report that mice with Atg16l1 deletion in intestinal epithelial cells (IECs) spontaneously develop transmural ileitis phenocopying ileal CD in an age-dependent manner, driven by the endoplasmic reticulum (ER) stress sensor IRE1α. IRE1α accumulates in Paneth cells of Atg16l1ΔIEC mice, and humans homozygous for ATG16L1T300A exhibit a corresponding increase of IRE1α in intestinal epithelial crypts. In contrast to a protective role of the IRE1β isoform, hyperactivated IRE1α also drives a similar ileitis developing earlier in life in Atg16l1;Xbp1ΔIEC mice, in which ER stress is induced by deletion of the unfolded protein response transcription factor XBP1. The selective autophagy receptor optineurin interacts with IRE1α, and optineurin deficiency amplifies IRE1α levels during ER stress. Furthermore, although dysbiosis of the ileal microbiota is present in Atg16l1;Xbp1ΔIEC mice as predicted from impaired Paneth cell antimicrobial function, such structural alteration of the microbiota does not trigger ileitis but, rather, aggravates dextran sodium sulfate–induced colitis. Hence, we conclude that defective autophagy in IECs may predispose to CD ileitis via impaired clearance of IRE1α aggregates during ER stress at this site.

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miR-802 regulates Paneth cell function and enterocyte differentiation in the mouse small intestine

Nature Communications ◽

10.1038/s41467-021-23298-3 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Algera Goga ◽

Büsra Yagabasan ◽

Karolin Herrmanns ◽

Svenja Godbersen ◽

Pamuditha N. Silva ◽

...

Keyword(s):

Small Intestine ◽

Epithelial Cell ◽

Intestinal Epithelium ◽

Cell Function ◽

Paneth Cell ◽

Cell Types ◽

Intestinal Epithelial ◽

Genetic Ablation ◽

Epithelial Homeostasis ◽

Enterocyte Differentiation

AbstractThe intestinal epithelium is a complex structure that integrates digestive, immunological, neuroendocrine, and regenerative functions. Epithelial homeostasis is maintained by a coordinated cross-talk of different epithelial cell types. Loss of integrity of the intestinal epithelium plays a key role in inflammatory diseases and gastrointestinal infection. Here we show that the intestine-enriched miR-802 is a central regulator of intestinal epithelial cell proliferation, Paneth cell function, and enterocyte differentiation. Genetic ablation of mir-802 in the small intestine of mice leads to decreased glucose uptake, impaired enterocyte differentiation, increased Paneth cell function and intestinal epithelial proliferation. These effects are mediated in part through derepression of the miR-802 target Tmed9, a modulator of Wnt and lysozyme/defensin secretion in Paneth cells, and the downstream Wnt signaling components Fzd5 and Tcf4. Mutant Tmed9 mice harboring mutations in miR-802 binding sites partially recapitulate the augmented Paneth cell function of mice lacking miR-802. Our study demonstrates a broad miR-802 network that is important for the integration of signaling pathways of different cell types controlling epithelial homeostasis in the small intestine.

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