Loss of Nckx3 Exacerbates Experimental DSS-Induced Colitis in Mice through p53/NF-κB Pathway

Inflammatory bowel diseases (IBDs) comprises a range of chronic inflammatory conditions of the intestinal tract. The incidence and prevalence of IBDs are increasing worldwide, but the precise etiology of these diseases is not completely understood. Calcium signaling plays a regulatory role in cellular proliferation. Nckx3, a potassium-dependent Na+/Ca2+ exchanger, is not only expressed in the brain but also in the aortic, uterine, and intestinal tissues, which contain abundant smooth muscle cells. This study investigated the role of Nckx3 in intestinal inflammation. Microarray analyses revealed the upregulation of the innate immune response-associated genes in the duodenum of Nckx3 knockout (KO) mice. The Nckx3 KO mice also showed an increase in IBD- and tumorigenesis-related genes. Using dextran sodium sulfate (DSS)-induced experimental colitis mice models, the Nckx3 KO mice showed severe colitis. Furthermore, the pathways involving p53 and NF-κB signaling were significantly upregulated by the absence of Nckx3. Overall, Nckx3 plays a critical role in the innate immune and immune response and may be central to the pathogenesis of IBD.

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Microbiota-independent immunological effects of non-digestible oligosaccharides in the context of inflammatory bowel diseases

Proceedings of The Nutrition Society ◽

10.1017/s0029665120006953 ◽

2020 ◽

Vol 79 (4) ◽

pp. 468-478 ◽

Cited By ~ 1

Author(s):

Stefania Del Fabbro ◽

Philip C. Calder ◽

Caroline E. Childs

Keyword(s):

Inflammatory Bowel Diseases ◽

Immune Cell ◽

Intestinal Inflammation ◽

Mechanisms Of Action ◽

Mucosal Inflammation ◽

Disease States ◽

Inflammatory Conditions ◽

Bowel Diseases ◽

Inflammatory Bowel

The aim of the present paper is to review the effects of non-digestible oligosaccharides (NDO) on immunity, focusing on their microbiota-independent mechanisms of action, as well as to explore their potential beneficial role in inflammatory bowel diseases (IBD). IBD are chronic, inflammatory conditions of the gastrointestinal tract. Individuals with IBD have an aberrant immune response to commensal microbiota, resulting in extensive mucosal inflammation and increased intestinal permeability. NDO are prebiotic fibres well known for their role in supporting intestinal health through modulation of the gut microbiota. NDO reach the colon intact and are fermented by commensal bacteria, resulting in the production of SCFA with immunomodulatory properties. In disease states characterised by increased gut permeability, prebiotics may also bypass the gut barrier and directly interact with intestinal and systemic immune cells, as demonstrated in patients with IBD and in infants with an immature gut. In vitro models show that fructooligosaccharides, inulin and galactooligosaccharides exert microbiota-independent effects on immunity by binding to toll-like receptors on monocytes, macrophages and intestinal epithelial cells and by modulating cytokine production and immune cell maturation. Moreover, animal models and human supplementation studies demonstrate that some prebiotics, including inulin and lactulose, might reduce intestinal inflammation and IBD symptoms. Although there are convincing preliminary data to support NDO as immunomodulators in the management of IBD, their mechanisms of action are still unclear and larger standardised studies need to be performed using a wider range of prebiotics.

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Myeloid-derived miR-223 regulates intestinal inflammation via repression of the NLRP3 inflammasome

Journal of Experimental Medicine ◽

10.1084/jem.20160462 ◽

2017 ◽

Vol 214 (6) ◽

pp. 1737-1752 ◽

Cited By ~ 139

Author(s):

Viola Neudecker ◽

Moritz Haneklaus ◽

Owen Jensen ◽

Ludmila Khailova ◽

Joanne C. Masterson ◽

...

Keyword(s):

Inflammatory Bowel Disease ◽

Nlrp3 Inflammasome ◽

Untranslated Region ◽

Intestinal Inflammation ◽

Experimental Colitis ◽

Innate Immune ◽

Mouse Line ◽

Preclinical Models ◽

Inflammatory Monocytes ◽

Inflammatory Bowel

MicroRNA (miRNA)-mediated RNA interference regulates many immune processes, but how miRNA circuits orchestrate aberrant intestinal inflammation during inflammatory bowel disease (IBD) is poorly defined. Here, we report that miR-223 limits intestinal inflammation by constraining the nlrp3 inflammasome. miR-223 was increased in intestinal biopsies from patients with active IBD and in preclinical models of intestinal inflammation. miR-223-/y mice presented with exacerbated myeloid-driven experimental colitis with heightened clinical, histopathological, and cytokine readouts. Mechanistically, enhanced NLRP3 inflammasome expression with elevated IL-1β was a predominant feature during the initiation of colitis with miR-223 deficiency. Depletion of CCR2+ inflammatory monocytes and pharmacologic blockade of IL-1β or NLRP3 abrogated this phenotype. Generation of a novel mouse line, with deletion of the miR-223 binding site in the NLRP3 3′ untranslated region, phenocopied the characteristics of miR-223-/y mice. Finally, nanoparticle-mediated overexpression of miR-223 attenuated experimental colitis, NLRP3 levels, and IL-1β release. Collectively, our data reveal a previously unappreciated role for miR-223 in regulating the innate immune response during intestinal inflammation.

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Cell Death and Inflammatory Bowel Diseases: Apoptosis, Necrosis, and Autophagy in the Intestinal Epithelium

BioMed Research International ◽

10.1155/2014/218493 ◽

2014 ◽

Vol 2014 ◽

pp. 1-12 ◽

Cited By ~ 39

Author(s):

Tiago Nunes ◽

Claudio Bernardazzi ◽

Heitor S. de Souza

Keyword(s):

Cell Death ◽

Inflammatory Bowel Diseases ◽

Intestinal Inflammation ◽

Critical Role ◽

Receptor Activation ◽

Tnf Receptor ◽

Programmed Necrosis ◽

Bowel Diseases ◽

Inflammatory Bowel ◽

Cell Death Mechanisms

Cell death mechanisms have been associated with the development of inflammatory bowel diseases in humans and mice. Recent studies suggested that a complex crosstalk between autophagy/apoptosis, microbe sensing, and enhanced endoplasmic reticulum stress in the epithelium could play a critical role in these diseases. In addition, necroptosis, a relatively novel programmed necrosis-like pathway associated with TNF receptor activation, seems to be also present in the pathogenesis of Crohn’s disease and in specific animal models for intestinal inflammation. This review attempts to cover new data related to cell death mechanisms and inflammatory bowel diseases.

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Identification and characterization of a novel Enterococcus bacteriophage with potential to ameliorate murine colitis

Scientific Reports ◽

10.1038/s41598-021-99602-4 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Junko Nishio ◽

Hideo Negishi ◽

Mika Yasui-Kato ◽

Shoji Miki ◽

Kazuhiko Miyanaga ◽

...

Keyword(s):

Intestinal Inflammation ◽

Experimental Colitis ◽

Murine Colitis ◽

Gut Homeostasis ◽

Bowel Diseases ◽

Mucin 2 ◽

Inflammatory Bowel ◽

Enterococcus Gallinarum ◽

Identification And Characterization

AbstractIncrease of the enteric bacteriophages (phage), components of the enteric virome, has been associated with the development of inflammatory bowel diseases. However, little is known about how a given phage contributes to the regulation of intestinal inflammation. In this study, we isolated a new phage associated with Enterococcus gallinarum, named phiEG37k, the level of which was increased in C57BL/6 mice with colitis development. We found that, irrespective of the state of inflammation, over 95% of the E. gallinarum population in the mice contained phiEG37k prophage within their genome and the phiEG37k titers were proportional to that of E. gallinarum in the gut. To explore whether phiEG37k impacts intestinal homeostasis and/or inflammation, we generated mice colonized either with E. gallinarum with or without the prophage phiEG37k. We found that the mice colonized with the bacteria with phiEG37k produced more Mucin 2 (MUC2) that serves to protect the intestinal epithelium, as compared to those colonized with the phage-free bacteria. Consistently, the former mice were less sensitive to experimental colitis than the latter mice. These results suggest that the newly isolated phage has the potential to protect the host by strengthening mucosal integrity. Our study may have clinical implication in further understanding of how bacteriophages contribute to the gut homeostasis and pathogenesis.

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A Dietary Cholesterol-Based Intestinal Inflammation Assay for Improving Drug-Discovery on Inflammatory Bowel Diseases

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.674749 ◽

2021 ◽

Vol 9 ◽

Author(s):

Nuno-Valério Silva ◽

Diogo Carregosa ◽

Catarina Gonçalves ◽

Otília V. Vieira ◽

Cláudia Nunes dos Santos ◽

...

Keyword(s):

Drug Discovery ◽

Inflammatory Bowel Diseases ◽

Intestinal Inflammation ◽

Feeding Strategy ◽

Critical Role ◽

Dietary Cholesterol ◽

High Cholesterol Diet ◽

Transgenic Zebrafish ◽

Bowel Diseases ◽

Inflammatory Bowel

Inflammatory bowel diseases (IBD) with chronic infiltration of immune cells in the gastrointestinal tract are common and largely incurable. The therapeutic targeting of IBD has been hampered by the complex causality of the disease, with environmental insults like cholesterol-enriched Western diets playing a critical role. To address this drug development challenge, we report an easy-to-handle dietary cholesterol-based in vivo assay that allows the screening of immune-modulatory therapeutics in transgenic zebrafish models. An improvement in the feeding strategy with high cholesterol diet (HCD) selectively induces a robust and consistent infiltration of myeloid cells in larvae intestines that is highly suitable for compound discovery efforts. Using transgenics with fluorescent reporter expression in neutrophils, we take advantage of the unique zebrafish larvae clarity to monitor an acute inflammatory response in a whole organism context with a fully functional innate immune system. The use of semi-automated image acquisition and processing combined with quantitative image analysis allows categorizing anti- or pro-inflammatory compounds based on a leukocytic inflammation index. Our HCD gut inflammation (HCD-GI) assay is simple, cost- and time-effective as well as highly physiological which makes it unique when compared to chemical-based zebrafish models of IBD. Besides, diet is a highly controlled, selective and targeted trigger of intestinal inflammation that avoids extra-intestinal outcomes and reduces the chances of chemical-induced toxicity during screenings. We show the validity of this assay for a screening platform by testing two dietary phenolic acids, namely gallic acid (GA; 3,4,5-trihydroxybenzoic acid) and ferulic acid (FA; 4-hydroxy-3-methoxycinnamic acid), with well described anti-inflammatory actions in animal models of IBD. Analysis of common IBD therapeutics (Prednisolone and Mesalamine) proved the fidelity of our IBD-like intestinal inflammation model. In conclusion, the HCD-GI assay can facilitate and accelerate drug discovery efforts on IBD, by identification of novel lead molecules with immune modulatory action on intestinal neutrophilic inflammation. This will serve as a jumping-off point for more profound analyses of drug mechanisms and pathways involved in early IBD immune responses.

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Dichotomous roles of neutrophils in modulating pathogenic and repair processes of inflammatory bowel diseases

Precision Clinical Medicine ◽

10.1093/pcmedi/pbab025 ◽

2021 ◽

Author(s):

Huimin Chen ◽

Xiaohan Wu ◽

Chunjin Xu ◽

Jian Lin ◽

Zhanju Liu

Keyword(s):

Immune Cells ◽

Inflammatory Bowel Diseases ◽

Intestinal Inflammation ◽

Critical Role ◽

Mucosal Inflammation ◽

Repair Processes ◽

Microbial Invasion ◽

Increased Risk ◽

Bowel Diseases ◽

Inflammatory Bowel

Abstract Neutrophils are considered as complex innate immune cells and play a critical role in maintaining intestinal mucosal homeostasis. They exert robust pro-inflammatory effects and recruit other immune cells in the acute phase of pathogen infection and intestinal inflammation, but paradoxically, they also limit exogenous microbial invasion and facilitate mucosal restoration. Hyperactivation or dysfunction of neutrophils results in abnormal immune responses, leading to multiple autoimmune and inflammatory diseases including systemic lupus erythematosus, rheumatoid arthritis, and inflammatory bowel diseases (IBD). As a refractory intestinal inflammatory disease, the pathogenesis and progression of IBD are associated with complicated immune response processes in which neutrophils are profoundly involved. However, the consensus on potential roles of neutrophils in modulating pathogenic and repair processes of IBD remains not fully understood. Accumulated infiltrating neutrophils cross the epithelial barrier and contribute to microbial dysbiosis, aggravated intestinal architectural damage, compromised resolution of intestinal inflammation and increased risk of thrombosis during IBD. Paradoxically, activated neutrophils are also associated with effective elimination of invaded microbiota, promoted angiogenesis and tissue restoration of gut mucosa in IBD. Here, we discuss the beneficial and detrimental roles of neutrophils in the onset and resolution of intestinal mucosal inflammation and provide a precise overview of neutrophil functions in the pathogenesis of IBD.

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Absence of GdX/UBL4A protects against inflammatory bowel diseases by regulating NF-κB signaling in DCs and macrophages

10.1101/376103 ◽

2018 ◽

Author(s):

Chunxiao Liu ◽

Yifan Zhou ◽

Mengdi Li ◽

Ying Wang ◽

Shigao Yang ◽

...

Keyword(s):

Immune Response ◽

Inflammatory Diseases ◽

Adaptor Protein ◽

Endotoxin Shock ◽

Innate Immune ◽

Mucosal Inflammation ◽

Innate Immune Responses ◽

Bowel Diseases ◽

Inflammatory Bowel ◽

Deficient Mice

AbstractNuclear factor-kappa B (NF-κB) activation is critical for innate immune responses. Here we report that the UBL4A (Ubiquitin-like protein 4A, also named GdX) enhances dendritic cells (DCs) and macrophages (Mφ)-mediated innate immune defenses by positively regulating NF-κB signaling. GdX-deficient mice were resistant to LPS-induced endotoxin shock and DSS-induced colitis. DC- or Mφ-specific GdX-deficient mice displayed alleviated mucosal inflammation, and the production of pro-inflammatory cytokines by GdX-deficient DCs and Mφ was reduced. Mechanistically, we found that PTPN2 (TC45) and PP2A form a complex with RelA (p65) to mediate its dephosphorylation whereas GdX interrupts the TC45/PP2A/p65 complex formation and restrict p65 dephosphorylation by trapping TC45. Our study provides a mechanism by which NF-κB signaling is positively regulated by an adaptor protein GdX in DC or Mφ to maintain the innate immune response. Targeting GdX could be a strategy to reduce over-activated immune response in inflammatory diseases.

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Protectin D1n-3 DPA and resolvin D5n-3 DPA are effectors of intestinal protection

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1617290114 ◽

2017 ◽

Vol 114 (15) ◽

pp. 3963-3968 ◽

Cited By ~ 68

Author(s):

Thomas Gobbetti ◽

Jesmond Dalli ◽

Romain A. Colas ◽

Donata Federici Canova ◽

Marius Aursnes ◽

...

Keyword(s):

Intestinal Inflammation ◽

Tissue Injury ◽

Leukocyte Adhesion ◽

Ischemia Reperfusion ◽

Experimental Colitis ◽

Protective Effects ◽

Inflammatory Conditions ◽

Bowel Diseases ◽

Affected Tissue ◽

Intestinal Ischemia Reperfusion

The resolution of inflammation is an active process orchestrated by specialized proresolving lipid mediators (SPM) that limit the host response within the affected tissue; failure of effective resolution may lead to tissue injury. Because persistence of inflammatory signals is a main feature of chronic inflammatory conditions, including inflammatory bowel diseases (IBDs), herein we investigate expression and functions of SPM in intestinal inflammation. Targeted liquid chromatography-tandem mass spectrometry-based metabololipidomics was used to identify SPMs from n-3 polyunsaturated fatty acids in human IBD colon biopsies, quantifying a significant up-regulation of the resolvin and protectin pathway compared with normal gut tissue. Systemic treatment with protectin (PD)1n-3 DPA or resolvin (Rv)D5n-3 DPA protected against colitis and intestinal ischemia/reperfusion-induced inflammation in mice. Inhibition of 15-lipoxygenase activity reduced PD1n-3 DPA and augmented intestinal inflammation in experimental colitis. Intravital microscopy of mouse mesenteric venules demonstrated that PD1n-3 DPA and RvD5n-3 DPA decreased the extent of leukocyte adhesion and emigration following ischemia-reperfusion. These data were translated by assessing human neutrophil–endothelial interactions under flow: PD1n-3 DPA and RvD5n-3 DPA reduced cell adhesion onto TNF-α–activated human endothelial monolayers. In conclusion, we propose that innovative therapies based on n-3 DPA-derived mediators could be developed to enable antiinflammatory and tissue protective effects in inflammatory pathologies of the gut.

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Mycobacterial Hsp65 antigen delivered by invasive Lactococcus lactis reduces intestinal inflammation and fibrosis in TNBS-induced chronic colitis model

Scientific Reports ◽

10.1038/s41598-020-77276-8 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Vanessa Pecini da Cunha ◽

Tatiane Melo Preisser ◽

Mariana Passos Santana ◽

Denise Carmona Cara Machado ◽

Vanessa Bastos Pereira ◽

...

Keyword(s):

Lactococcus Lactis ◽

Intestinal Inflammation ◽

Experimental Colitis ◽

Trinitrobenzene Sulfonic Acid ◽

Intestinal Fibrosis ◽

Inflammatory Damage ◽

Bowel Diseases ◽

Inflammatory Bowel ◽

Shock Proteins ◽

Chronic Intestinal Inflammation

AbstractIntestinal fibrosis associated with Crohn’s disease (CD), which a common and serious complication of inflammatory bowel diseases. In this context, heat shock proteins (HSPs) might serve as an alternative treatment because these antigens play important roles in the regulation of effector T cells. We thus evaluated the anti-inflammatory and antifibrotic capacities of an invasive and Hsp65-producing strain—Lactococcus lactis NCDO2118 FnBPA+ (pXYCYT:Hsp65)—in chronic intestinal inflammation to assess its potential as an alternative therapeutic strategy against fibrotic CD. Experimental colitis was induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS) in BALB/c mice, and the mice were treated orally with L. lactis NCDO2118 FnBPA+ (pXYCYT:Hsp65) via intragastric gavage. The oral administration of this strain significantly attenuated the severity of inflammation and intestinal fibrosis in mice (p < 0.05). These results are mainly justified by reductions in the levels of the pro-fibrotic cytokines IL-13 and TGF-β and increases in the concentration of the regulatory cytokine IL-10. The L. lactis NCDO2118 FnBPA+ (pXYCYT:Hsp65) strain contributed to reductions in the severity of inflammatory damage in chronic experimental CD, and these findings confirm the effectiveness of this new antifibrotic strategy based on the delivery of therapeutic proteins to inside cells of the host intestinal mucosa.

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Mitochondria and Inflammatory Bowel Diseases: Toward a Stratified Therapeutic Intervention

Annual Review of Physiology ◽

10.1146/annurev-physiol-060821-083306 ◽

2021 ◽

Vol 84 (1) ◽

Author(s):

Gwo-tzer Ho ◽

Arianne L. Theiss

Keyword(s):

Mitochondrial Dysfunction ◽

Intestinal Inflammation ◽

Critical Role ◽

Annual Review ◽

Publication Date ◽

Cellular Functions ◽

Targeted Interventions ◽

Bowel Diseases ◽

Key Factor ◽

Inflammatory Bowel

Mitochondria serve numerous critical cellular functions, rapidly responding to extracellular stimuli and cellular demands while dynamically communicating with other organelles. Mitochondrial function in the gastrointestinal epithelium plays a critical role in maintaining intestinal health. Emerging studies implicate the involvement of mitochondrial dysfunction in inflammatory bowel disease (IBD). This review presents mitochondrial metabolism, function, and quality control that converge in intestinal epithelial stemness, differentiation programs, barrier integrity, and innate immunity to influence intestinal inflammation. Intestinal and disease characteristics that set the stage for mitochondrial dysfunction being a key factor in IBD, and in turn, pathogenic mitochondrial mechanisms influencing and potentiating the development of IBD, are discussed. These findings establish the basis for potential mitochondrial-targeted interventions for IBD therapy. Expected final online publication date for the Annual Review of Physiology, Volume 84 is February 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

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