Expression Of Inflammatory Bowel Disease Research Articles

Exploring Microbial Metabolite Receptors in Inflammatory Bowel Disease: An In Silico Analysis of Their Potential Role in Inflammation and Fibrosis.

Metabolites produced by dysbiotic intestinal microbiota can influence disease pathophysiology by participating in ligand-receptor interactions. Our aim was to investigate the differential expression of metabolite receptor (MR) genes between inflammatory bowel disease (IBD), healthy individuals (HIs), and disease controls in order to identify possible interactions with inflammatory and fibrotic pathways in the intestine. RNA-sequencing datasets containing 643 Crohn's disease (CD) patients, 467 ulcerative colitis (UC) patients and 295 HIs, and 4 Campylobacter jejuni-infected individuals were retrieved from the Sequence Read Archive, and differential expression was performed using the RaNA-seq online platform. The identified differentially expressed MR genes were used for correlation analysis with up- and downregulated genes in IBD, as well as functional enrichment analysis using a R based pipeline. Overall, 15 MR genes exhibited dysregulated expression in IBD. In inflamed CD, the hydroxycarboxylic acid receptors 2 and 3 (HCAR2, HCAR3) were upregulated and were associated with the recruitment of innate immune cells, while, in the non-inflamed CD ileum, the cannabinoid receptor 1 (CNR1) and the sphingosine-1-phospate receptor 4 (S1PR4) were downregulated and were involved in the regulation of B-cell activation. In inflamed UC, the upregulated receptors HCAR2 and HCAR3 were more closely associated with the process of TH-17 cell differentiation, while the pregnane X receptor (NR1I2) and the transient receptor potential vanilloid 1 (TRPV1) were downregulated and were involved in epithelial barrier maintenance. Our results elucidate the landscape of metabolite receptor expression in IBD, highlighting associations with disease-related functions that could guide the development of new targeted therapies.

"Decoding inflammation: glycoprotein a repetition predominant, microRNA-142-3-p, and metastasis associated lung adenocarcinoma transcript 1: as novel inflammatory biomarkers of inflammatory bowel disease".

Inflammatory bowel disease (IBD) is a chronic gastrointestinal (GI) condition comprising Crohn's disease (CD) and ulcerative colitis (UC). The pathogenesis involves immune system dysregulation, with increased Th (T helper cell)17 cells and reduced regulatory T cell (Treg) differentiation. Transforming growth factor-β (TGF-β) secretion from Tregs helps control inflammation, and its production is regulated by glycoprotein-A repetition predominant (GARP) protein along with non-coding RNAs (ncRNAs) like microRNA(miR)-142-3p and metastasis associated lung adenocarcinoma transcript 1 (MALAT1) long non-coding RNAs (LncRNAs). This study analyzed their expression in IBD. Blood samples were collected from 44 IBD patients, and 22 healthy controls (HC). RNA extraction and circular DNA (cDNA) synthesis were performed. Real-time polymerase chain reaction (RT-PCR) measured gene expression of GARP, MALAT1, and miR-142-3p. Correlations and group differences were statistically analyzed. Compared to controls, GARP was downregulated while MALAT1 and miR-142-3p were upregulated significantly in IBD group. GARP and MALAT1 expressions positively correlated in controls. MALAT1 and miR-142-3p expressions positively correlated in IBD group. MALAT1 was downregulated in aged HC but upregulated with smoking history across groups. No correlations occurred between gene expression and gender, diet, infections, or disease activity scores. Dysregulation of GARP, MALAT1, and miR-142-3p likely contributes to inflammation in IBD by reducing TGF-β. MALAT1 is linked to smoking and age-related changes. These genes have potential as diagnostic markers or therapeutic targets for personalized IBD treatment.

P379 PD-1-positive cells contribute to the diagnosis of Inflammatory Bowel Disease and can aid in predicting response to vedolizumab

Abstract Background Differentiating inflammatory bowel disease (IBD) from other inflammatory diseases is often challenging. Programmed cell death protein-1 (PD-1) is expressed in T cells and is an indicator of their exhaustion. The role of PD-1 expression in diagnosing IBD and predicting the response of biologic agents remains inconclusive. Methods In this study, endoscopic biopsy samples of 19 patients diagnosed with IBD and other inflammatory diseases were analysed using multiplexed immunohistochemistry . Other inflammatory diseases comprised five patients with intestinal tuberculosis, and five with intestinal Behçet’s disease. Additionally, a separate prospectively maintained "vedolizumab (VDZ) cohort", established in 12 ulcerative colitis(UC) patients who underwent endoscopic biopsy before VDZ administration was analysed to predict response to VDZ. Results In the immunohistochemistry analysis, the cell density of T cell subsets, including helper T cell (Th), cytotoxic T cell (Tc), regulatory T cell (Treg), PD-1+ cell, PD-1+ Th, PD-1+ Tc, and PD-1+ Treg was investigated and compared between IBD and other inflammatory disease. Cell densities of PD-1+ cells (p=0.028), PD-1+ Th (p=0.008), and PD-1+ Treg (p=0.024) were higher in IBD compared with other inflammatory disease.(Figure 1) In addition, the proportion of PD-1+ cells in Th and Treg was higher in the IBD group than in the other inflammatory disease group (median 7% vs. 3%, p=0.008; median 6% vs. 2%, p=0.004, respectively). Comparing within IBD, the cell density of Treg was higher in Crohn’s disease(CD) than in UC (p=0.042), whilst the cell density of Tc was higher in CD than in UC with marginal statistical significance (p=0.066). In the VDZ cohort, patients with a 14-week steroid-free clinical response had higher levels of PD-1+ cells (p=0.026), PD-1+ Th (p=0.026), and PD-1+ Treg (p=0.041) than the no response group.(Table 1) Conclusion We explored the relationship between PD-1 expression and IBD using multiplexed immunohistochemistry. We compared IBD and other inflammatory disease in their expression of various immune cells including PD-1+ cells, and found IBD had higher PD-1 expression, as well as higher PD-1+ Th and PD-1+ Treg compared with other inflammatory diseases. In addition, VDZ-responsive patients had higher PD-1 expression. Our study confirmed the difference in PD-1 expression in IBD and other inflammatory diseases and revealed that PD-1 expression could be a predictive marker for VDZ responsiveness.

Faecalibacterium prausnitzii promotes intestinal epithelial IL-18 production through activation of the HIF1α pathway.

Intestinal epithelial cells produce interleukin-18 (IL-18), a key factor in promoting epithelial barrier integrity. Here, we analyzed the potential role of gut bacteria and the hypoxia-inducible factor 1α (HIF1α) pathway in regulating mucosal IL18 expression in inflammatory bowel disease (IBD). Mucosal samples from patients with IBD (n = 760) were analyzed for bacterial composition, IL18 levels and HIF1α pathway activation. Wild-type Caco-2 and CRISPR/Cas9-engineered Caco-2-HIF1A-null cells were cocultured with Faecalibacterium prausnitzii in a "Human oxygen-Bacteria anaerobic" in vitro system and analyzed by RNA sequencing. Mucosal IL18 mRNA levels correlated positively with the abundance of mucosal-associated butyrate-producing bacteria, in particular F. prausnitzii, and with HIF1α pathway activation in patients with IBD. HIF1α-mediated expression of IL18, either by a pharmacological agonist (dimethyloxallyl glycine) or F. prausnitzii, was abrogated in Caco-2-HIF1A-null cells. Butyrate-producing gut bacteria like F. prausnitzii regulate mucosal IL18 expression in a HIF1α-dependent manner that may aid in mucosal healing in IBD.

Upregulated Tβ4 expression in inflammatory bowel disease impairs the intestinal mucus barrier by inhibiting autophagy in mice

Disrupted intestinal barrier homeostasis is fundamental to inflammatory bowel disease. Thymosin β4 (Tβ4) improves inflammation and has beneficial effects in dry-eye diseases, but its effects on the intestinal mucus barrier remain unknown. Therefore, this study evaluated the underlying regulatory mechanisms and effects of Tβ4 by examining Tβ4 expression in a mouse model with dextran sodium sulfate (DSS)-induced colitis and colonic barrier damage. Additionally, we intraperitoneally injected C57BL/6 mice with Tβ4 to assess barrier function, microtubule-associated protein 1 light chain 3 (LC3II) protein expression, and autophagy. Finally, normal human colon tissue and colon carcinoma cells (Caco2) were cultured to verify Tβ4-induced barrier function and autophagy changes. Mucin2 levels decreased, microbial infiltration increased, and Tβ4 expression increased in the colitis mouse model versus the control mice, indicating mucus barrier damage. Moreover, Tβ4-treated C57BL/6 mice had damaged intestinal mucus barriers and decreased LC3II levels. Tβ4 also inhibited colonic mucin2 production, disrupted tight junctions, and downregulated autophagy; these results were confirmed in Caco2 cells and normal human colon tissue. In summary, Tβ4 may be implicated in colitis by compromising the integrity of the intestinal mucus barrier and inhibiting autophagy. Thus, Tβ4 could be a new diagnostic marker for intestinal barrier defects.

Loss of claudin-3 expression increases colitis risk by promoting Gut Dysbiosis

ABSTRACT Dysregulation of both the gut barrier and microbiota (dysbiosis) promotes susceptibility to and severity of Inflammatory Bowel Diseases (IBD). Leaky gut and dysbiosis often coexist; however, potential interdependence and molecular regulation are not well understood. Robust expression of claudin-3 (CLDN3) characterizes the gut epithelium, and studies have demonstrated a positive association between CLDN3 expression and gut barrier maturity and integrity, including in response to probiotics. However, the exact status and causal role of CLDN3 in IBD and regulation of gut dysbiosis remain unknown. Analysis of mouse and human IBD cohorts helped examine CLDN3 expression in IBD. The causal role was determined by modeling CLDN3 loss of expression during experimental colitis. 16S sequencing and in silico analysis helped examine gut microbiota diversity between Cldn3KO and WT mice and potential host metabolic responses. Fecal microbiota transplant (FMT) studies were performed to assess the role of gut dysbiosis in the increased susceptibility of Cldn3KO mice to colitis. A significant decrease in CLDN3 expression characterized IBD and CLDN3 loss of expression promoted colitis. 16S sequencing analysis suggested gut microbiota changes in Cldn3KO mice that were capable of modulating fatty acid metabolism and oxidative stress response. FMT from naïve Cldn3KO mice promoted colitis susceptibility in recipient germ-free mice (GFM) compared with GFM-receiving microbiota from WT mice. Our data demonstrate a critical role of CLDN3 in maintaining normal gut microbiota and inflammatory responses, which can be harnessed to develop novel therapeutic opportunities for patients with IBD.

Decoding the IBD paradox: A triadic interplay between REG3, enterococci, and NOD2

In this issue of Cell Host & Microbe, Jang etal. decode an intriguing paradox: excessive antimicrobial peptide-regenerating family member 3 (REG3) expression in inflammatory bowel disease (IBD) patients depletes protective enterococci. This depletion impairs the NOD2 anti-inflammatory pathway and perpetuates inflammation, thus providing insight into IBD pathogenesis.

Characterization of canine intestinal microRNA expression in inflammatory bowel disease and T-cell lymphoma

Differentiating between canine inflammatory bowel disease (IBD) and intestinal T-cell lymphoma by histopathological examination of endoscopically-derived intestinal biopsies can be challenging and involves an invasive procedure requiring specialized equipment and training. A rapid, non-invasive method of diagnosis, such as blood or faecal analysis for a conserved and stable biomarker, would be a useful adjunct or replacement. Studies on dogs and humans with various types of lymphoma have shown altered microRNA (miRNA) expression patterns in blood, faeces and tissues indicating their potential use as biomarkers of disease. The present study used residual archived endoscopically-derived, formalin-fixed, paraffin-embedded (FFPE) duodenal tissue taken from pet dogs undergoing routine investigation of gastrointestinal disease. The dogs had previously been diagnosed with either normal/minimal intestinal inflammation, severe IBD or intestinal T-cell lymphoma. Next generation sequencing with qPCR validation was used to elucidate differentially expressed miRNAs between groups. Our results show that miRNA can be extracted from archived endoscopically-derived FFPE tissues from the canine duodenum and used to differentiate normal/minimally inflamed canine duodenal tissue from severe lymphoplasmacytic IBD and T-cell lymphoma.

Loss of Claudin-3 Impairs Intestinal Barrier Function and Gut Microbiota to Promote Colitis

Objective: Inflammatory Bowel Disease (IBD) is multifactorial autoimmune disorder where dysregulation of the gut barrier and gut microbiota (Dysbiosis) play critical role in disease onset and progression. A leaky gut often coexists with the dysbiotic gut microbiota, however a pathobiological integration is not well understood. Intestinal claudin-3 expression is robust, and we have reported that its loss promotes gut permeability and colon cancer. Increased gut permeability and colon cancer risk characterize Inflammatory Bowel Disease (IBD). The current study was undertaken to delineate the status and role of claudin-3 expression in IBD. Methods: Multiple murine models of colitis and IBD patient biopsy were used. Claudin-3KO mice were used to determine causal role. Multiplex ELISA, RNAseq and 16S DNA sequencing helped determine inflammatory cytokines, transcriptomic changes, and gut microbiota diversity, respectively. Fecal microbiota transplantation (FMT) and antibiotic treatment studies were done to test the role of claudin-3/gut dysbiosis axis in promoting IBD. Results: A significant decrease in claudin-3 expression was observed in IBD patient samples or the colon of mice subjected to DSS- (2.5% w/v) or C. rodentium-colitis (5 x108 CFU/oral). colitis (p<0.001). To test the causal role, claudin-3KO mice were subjected to DSS-colitis (acute and chronic), and infectious colitis. A significant change in the body weight, colon thickness, and mucosal injury score (p<0.001) characterized the colitis challenged claudin-3KO versus WT mice. P-Stat3 expression was highly upregulated in colitis challenged claudin-3KO mice. Interestingly, administration of an antibiotic cocktail rescued the colitis severity in claudin-3KO mice. The 16s metagenomics further revealed that the gut microbiota of claudin-3KO mice is diverse and clustered distinctly from the WT mice. To determine if gut dysbiosis in claudin-3KO mice promotes susceptibility to colitis, we performed FMT from naïve Claudin-3KO mice to germ-free WT mice. When subjected to DSS-colitis, germ-free mice transplanted with Claudin3-KO gut microbiota showed significantly higher colitis severity versus WT (p<0.05). Conclusion: We summarize that the loss of colonic Cldn3 deregulates mucosal barrier integrity to induce gut dysbiosis and susceptibility to IBD. We propose that claudin-3 can be a novel therapeutic target in strengthening the gut barrier integrity to diminish IBD susceptibility. National Institute of Health, Veterans Administration This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Correlations between the symptoms of insomnia, depression, sleep quality, antitumor necrosis factor therapy, and disrupted circadian clock gene expression in inflammatory bowel disease.

Inflammatory bowel disease (IBD) might be accompanied by emotional disturbances. Circadian rhythm genes, such as brain and muscle ARNT‑Like 1 (BMAL1), circadian locomotor output cycles kaput (CLOCK), neuronal PAS domain protein 2 (NPAS2), or nuclear receptor subfamily 1 group D member 1 (NR1D1) are related to inflammation and psychiatric symptoms that might modulate their expression. The study aimed to compare the expression of the BMAL1, CLOCK, NPAS2, NR1D1 mRNA in IBD patients and healthy controls (HCs). We evaluated the association between the gene expression and the disease severity, antitumor necrosis factor (TNF) therapy, sleep quality, insomnia, and depression. A total of 81 IBD patients and 44 HCs were recruited and classified according to the disease activity and IBD type (ulcerative colitis [UC] or Crohn disease [CD]). The participants filled out questionnaires assessing their sleep quality, daytime sleepiness, insomnia, and depression. Venous blood samples were collected, and the IBD patients on the anti‑TNF therapy had their blood drawn before and after 14 weeks of the treatment. In comparison with HCs, the IBD group had decreased expression of all studied genes apart from the BMAL1 gene. UC individuals with exacerbation had decreased expression of the CLOCK and the NPAS2 genes, as compared with the remission group. UC severity negatively correlated with the CLOCK, NPAS2, and NR1D1 mRNA levels. The IBD participants with depression symptoms had a decreased expression of the CLOCK and the NR1D1 genes, as compared with those without mood disturbances. Poor sleep quality was associated with a decreased expression of the NR1D1 gene. Biologic treatment decreased the expression of the BMAL1 gene. Disruption of the clock gene expression might constitute a molecular background of sleep disorders and depression in IBD and might contribute to UC exacerbation.

IL-22-Activated MUC13 Impacts on Colonic Barrier Function through JAK1/STAT3, SNAI1/ZEB1 and ROCK2/MAPK Signaling.

Overexpression of the transmembrane mucin MUC13, as seen in inflammatory bowel diseases (IBD), could potentially impact barrier function. This study aimed to explore how inflammation-induced MUC13 disrupts epithelial barrier integrity by affecting junctional protein expression in IBD, thereby also considering the involvement of MUC1. RNA sequencing and permeability assays were performed using LS513 cells transfected with MUC1 and MUC13 siRNA and subsequently stimulated with IL-22. In vivo intestinal permeability and MUC13-related signaling pathways affecting barrier function were investigated in acute and chronic DSS-induced colitis wildtype and Muc13-/- mice. Finally, the expression of MUC13, its regulators and other barrier mediators were studied in IBD and control patients. Mucin knockdown in intestinal epithelial cells affected gene expression of several barrier mediators in the presence/absence of inflammation. IL-22-induced MUC13 expression impacted barrier function by modulating the JAK1/STAT3, SNAI1/ZEB1 and ROCK2/MAPK signaling pathways, with a cooperating role for MUC1. In response to DSS, MUC13 was protective during the acute phase whereas it caused more harm upon chronic colitis. The pathways accounting for the MUC13-mediated barrier dysfunction were also altered upon inflammation in IBD patients. These novel findings indicate an active role for aberrant MUC13 signaling inducing intestinal barrier dysfunction upon inflammation with MUC1 as collaborating partner.

MiRNA-21 Regulates Multiple Factors in the Pathogenesis of IBD: A Literature Review

Introduction: Inflammatory Bowel Disease (IBD) is characterized by chronic inflammation in the gastrointestinal tract (GIT) via dysbiosis of the gut microbiome and weakening of the epithelial and mucosal barriers. Although the causative nature of IBD remains unknown, several studies have demonstrated that aberrant host-microRNA (miRNA) activity contributes to its pathophysiology. For example, miRNA-21 contributes to IBD through three mechanisms. Firstly, by increasing gut permeability via the ARF4 pathway. Secondly, by decreasing gut mucosal secretions via interfering with host goblet cells. Finally, by regulating proteins that inhibit host autophagy and decreasing immune response via the Phosphatase and Tensin Homolog (PTEN) and Akt pathway. The proposed study aims to answer the following question: how does aberrant expression of miRNA-21 in IBD contribute to the disease? Methods: This review highlights and summarizes relevant studies on the miRNA-21 regulation of key pathways in the pathogenesis of IBD. Searches used electronic databases including PubMed, and Google Scholar for keywords such as “Inflammatory bowel disease” and “miRNA-21” and other additional relevant terms from years 2016 to 2022. Review papers that met our criteria and the relevant papers they referenced, regardless of their publication date, were manually searched for. Figures were made in part using KeyNote and Serveir Medical Art. Discussion: Intracellular pathways contribute to chronic inflammation in IBD such as the PTEN pathway and pro-inflammatory cytokines like TNF-α. These pathways have been shown to influence the mucosal barrier, epithelial barrier, and the immune system in the gut. These pathways are regulated by miRNA-21, demonstrating miRNA-21 as a key regulator in IBD. Both PTEN and TNF-α also contribute to levels of angiogenesis in the gut through the regulation of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor (HIF), further demonstrating the intricacies of the miRNA-21 pathway regulation in IBD. Conclusion: The research highlighted in this review provides insight into the mechanisms of aberrant miRNA expression in IBD. Furthermore, knowledge of such molecular mechanisms has clinical and research applications, including identifying diagnostic biomarkers, less invasive screening techniques, and novel drug therapies.

MiRNA-21 Regulates Multiple Factors in the Pathogenesis of IBD: A Literature Review

Introduction: Inflammatory Bowel Disease (IBD) is characterized by chronic inflammation in the gastrointestinal tract (GIT) via dysbiosis of the gut microbiome and weakening of the epithelial and mucosal barriers. Although the causative nature of IBD remains unknown, several studies have demonstrated that aberrant host-microRNA (miRNA) activity contributes to its pathophysiology. For example, miRNA-21 contributes to IBD through three mechanisms. Firstly, by increasing gut permeability via the ARF4 pathway. Secondly, by decreasing gut mucosal secretions via interfering with host goblet cells. Finally, by regulating proteins that inhibit host autophagy and decreasing immune response via the Phosphatase and Tensin Homolog (PTEN) and Akt pathway. The proposed study aims to answer the following question: how does aberrant expression of miRNA-21 in IBD contribute to the disease? Methods: This review highlights and summarizes relevant studies on the miRNA-21 regulation of key pathways in the pathogenesis of IBD. Searches used electronic databases including PubMed, and Google Scholar for keywords such as “Inflammatory bowel disease” and “miRNA-21” and other additional relevant terms from years 2016 to 2022. Review papers that met our criteria and the relevant papers they referenced, regardless of their publication date, were manually searched for. Figures were made in part using KeyNote and Serveir Medical Art. Discussion: Intracellular pathways contribute to chronic inflammation in IBD such as the PTEN pathway and pro-inflammatory cytokines like TNF-α. These pathways have been shown to influence the mucosal barrier, epithelial barrier, and the immune system in the gut. These pathways are regulated by miRNA-21, demonstrating miRNA-21 as a key regulator in IBD. Both PTEN and TNF-α also contribute to levels of angiogenesis in the gut through the regulation of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor (HIF), further demonstrating the intricacies of the miRNA-21 pathway regulation in IBD. Conclusion: The research highlighted in this review provides insight into the mechanisms of aberrant miRNA expression in IBD. Furthermore, knowledge of such molecular mechanisms has clinical and research applications, including identifying diagnostic biomarkers, less invasive screening techniques, and novel drug therapies.

A distal super-enhancer activates oncogenic ETS2 via recruiting MECOM in inflammatory bowel disease and colorectal cancer

Abnormal activities of distal cis-regulatory elements (CREs) contribute to the initiation and progression of cancer. Gain of super-enhancer (SE), a highly active distal CRE, is essential for the activation of key oncogenes in various cancers. However, the mechanism of action for most tumor-specific SEs still largely remains elusive. Here, we report that a candidate oncogene ETS2 was activated by a distal SE in inflammatory bowel disease (IBD) and colorectal cancer (CRC). The SE physically interacted with the ETS2 promoter and was required for the transcription activation of ETS2. Strikingly, the ETS2-SE activity was dramatically upregulated in both IBD and CRC tissues when compared to normal colon controls and was strongly correlated with the level of ETS2 expression. The tumor-specific activation of ETS2-SE was further validated by increased enhancer RNA transcription from this region in CRC. Intriguingly, a known IBD-risk SNP resides in the ETS2-SE and the genetic variant modulated the level of ETS2 expression through affecting the binding of an oncogenic transcription factor MECOM. Silencing of MECOM induced significant downregulation of ETS2 in CRC cells, and the level of MECOM and ETS2 correlated well with each other in CRC and IBD samples. Functionally, MECOM and ETS2 were both required for maintaining the colony-formation and sphere-formation capacities of CRC cells and MECOM was crucial for promoting migration. Taken together, we uncovered a novel disease-specific SE that distantly drives oncogenic ETS2 expression in IBD and CRC and delineated a mechanistic link between non-coding genetic variation and epigenetic regulation of gene transcription.

Increased stromal PFKFB3-mediated glycolysis in inflammatory bowel disease contributes to intestinal inflammation

Inflammatory bowel disease (IBD) is a chronic relapsing inflammation of the intestinal tract with currently not well-understood pathogenesis. In addition to the involvement of immune cells, increasing studies show an important role for fibroblasts in the pathogenesis of IBD. Previous work showed that glycolysis is the preferred energy source for fibroblasts in fibrotic diseases. 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase 3 (PFKFB3) is a key kinase supporting glycolysis. Increased expression of PFKFB3 in several cancers and inflammatory diseases has been previously reported, but the metabolic status of fibroblasts and the role of PFKFB3 in patients with IBD are currently unknown. Therefore, in this study, we evaluated the role of glycolysis and PFKFB3 expression in IBD. Single-sample gene set enrichment analysis (ssGSEA) revealed that glycolysis was significantly higher in IBD intestinal samples, compared to healthy controls, which was confirmed in the validation cohorts of IBD patients. Single-cell sequencing data indicated that PFKFB3 expression was higher in IBD-derived stromal cells. In vitro, PFKFB3 expression in IBD-derived fibroblasts was increased after the stimulation with pro-inflammatory cytokines. Using seahorse real-time cell metabolic analysis, inflamed fibroblasts were shown to have a higher extracellular acidification rate and a lower oxygen consumption rate, which could be reversed by inhibition of JAK/STAT pathway. Furthermore, increased expression of pro-inflammatory cytokines and chemokines in fibroblasts could be reverted by PFK15, a specific inhibitor of PFKFB3. In vivo experiments showed that PFK15 reduced the severity of dextran sulfate sodium (DSS)- and Tcell transfer induced colitis, which was accompanied by a reduction in immune cell infiltration in the intestines. These findings suggest that increased stromal PFKFB3 expression contributes to inflammation and the pathological function of fibroblasts in IBD. Inhibition of PFKFB3 suppressed their inflammatory characteristics.

NF-kappa B expression in resected specimen of colonic cancer is higher compared to its expression in inflammatory bowel diseases and polyps

NF-Kappa B has a significant role in inflammatory processes as well as in colorectal cancer. The aim of this study was to compare the expression of NF-kappa B in colonic adenocarcinoma specimen, colonic adenomas and inflammatory colonic tissues. Patients with colorectal cancer (CRC), colonic adenomas and inflammatory processes undergoing surgery were recruited. Following a routine pathological evaluation tissue samples were stained using anti NF-κB monoclonal antibodies. Expression of NF-κB was quantified using IMAGEJ program for immunohistochemistry staining. Samples were also stained and quantified for CEA expression. Fifty-six patients were included. 30 cancers, 6 polyps and 20 inflammatory processes. Expression of NF-κB was similar between polypoid and inflammation etiologies. However, it was significantly higher in CRC compared to both (p < 0.05). In cancer patients, NF-κB expression in the resection margins was correlated with positive node status. CEA expression was higher in the cancer group, less in the IBD group and the lowest in the colonic non diseased margins. Our results provide a supportive evidence that NF-κB pathway is strongly involved in colon cancer development and metastasis. Interestingly, expression of NF-κB in benign polypoid lesions was as high as in inflammatory etiologies. This support the role of NF-κB early in the adenoma to carcinoma sequence. Further research is needed to evaluate the exact role of NF-κB in tumor progression in order to look for diagnostic and therapeutic possibilities.

Intramucosal Calprotectin Expression in Inflammatory Bowel Disease (IBD) and Non-IBD Colorectal Inflammation

BACKGROUND: Inflammatory bowel disease (IBD) diagnosis remains a challenge accompanied with high numbers of misdiagnosis causing suboptimal management. Tons of trials have been conducted to improve the diagnostic accuracy, one of which is the use of biomarker such as calprotectin. Calprotectin can be detected in tissue (intramucosal) and is becoming a potential marker of IBD. AIM: This study aims to determine intramucosal calprotectin expression in IBD, non-IBD colitis, and control. METHODS: This analytic retrospective study included consecutively sampled IBD and non-IBD colitis colorectal biopsy specimens, and control group obtained from Cipto Mangunkusumo Hospital registered from 2017 to 2019. Cases were included in the study if specimens were indicative of IBD and non-IBD clinically and histopathologically and no abnormality were found histopathologically in the control group. Specimens with non-adequate data from the hospital medical records or with missing tissue slides were excluded from the study. Calprotectin immunostaining was conducted to evaluate mean intramucosal calprotectin expression (cell/HPF) in each group. RESULTS: Most of the samples from IBD and non-IBD group (45 samples each) showed mild active inflammation. Mucosal calprotectin expression in aforementioned groups was higher than that of control group (p &lt; 0.001). Subjects with active inflammation showed higher calprotectin expression compared to those with inactive inflammation (p &lt; 0.001). Calprotectin expression was also related to activity grade. CONCLUSION: Higher calprotectin expression showed significant association with the presence of inflammation and disease activity. However, the application of intramucosal calprotectin immunohistochemistry test to determine inflammatory etiology (IBD vs. non-IBD) still needs to be further evaluated.

Interleukin-26 Expression in Inflammatory Bowel Disease and Its Immunoregulatory Effects on Macrophages.

Background and AimInterleukin-26 (IL-26) has been implicated in several chronic inflammatory diseases. However, its role in inflammatory bowel disease (IBD) remains to be elucidated. We aimed to investigate IL-26 expression in IBD and its immunoregulatory effects on macrophages.MethodsWe assessed IL-26 expression in the intestinal mucosa and blood samples of IBD patients and healthy controls (HC). The associations between the clinical characteristics of IBD and IL-26 expression levels in serum and peripheral blood mononuclear cells (PBMCs) were investigated. In addition, the transcriptional changes in THP-1 macrophages exposed to IL-26 were determined by RNA sequencing and validated with qRT-PCR, ELISA and western blots.ResultsCompared with HC, in IBD patients, IL-26 expression levels were elevated in the inflamed intestinal mucosa, and reduced in serum and PBMCs. IL-26 mRNA levels in PBMCs, but not serum IL-26 levels, were inversely correlated with disease activity in IBD. Furthermore, IL-26 mRNA levels in PBMCs were significantly lower in patients with complicated Crohn’s disease. A total of 1,303 differentially expressed protein-coding genes were identified between untreated and IL-26-treated macrophages. The up-regulated genes showed enrichment in some inflammatory and immune-related processes and pathways. Additionally, GSEA showed that neutrophil, monocyte, and lymphocyte chemotaxis was significantly enriched in IL-26-treated macrophages. Further validation revealed that IL-26 promotes the secretion of multiple inflammatory cytokines and chemokines and upregulates the expression of adhesion molecules, MMP-8, and MMP-9 while inhibiting MMP-1 in macrophages.ConclusionCompared with HC, in IBD patients, IL-26 levels were elevated in the inflamed intestinal mucosa, and reduced in the peripheral blood. The transcriptional changes in macrophages exposed to IL-26 suggest that IL-26 may amplify the aberrant immune response in IBD by activating macrophages.

MicroRNAs as potential biomarkers for the diagnosis of inflammatory bowel disease: a systematic review and meta-analysis.

ObjectiveThe clinical importance of aberrantly expressed microRNAs (miRNAs) in diagnosing inflammatory bowel disease (IBD) has not been well established, so was investigated in this systematic review and meta-analysis.MethodsArticles in online databases from inception to March 17, 2021 were retrieved. Random effects meta-analysis was used to obtain sensitivity, specificity, positive (PLRs) and negative likelihood ratios (NLRs), diagnostic odds ratios (DORs), and areas under the curve (AUC) with 95% confidence intervals (CI) for IBD diagnosis.ResultsOf 117 studies reporting altered miRNA expression in IBD included in the systematic review, 15 involving 937 patients with IBD and 707 controls, 22 miRNAs, and two miRNA panels were eligible for meta-analysis. Pooled analyses showed a moderate diagnostic accuracy for miRNAs in the IBD diagnosis, with a sensitivity of 0.80 (95% CI: 0.79–0.82), specificity of 0.84 (95% CI: 0.82–0.86), DOR of 21.19 (95% CI: 13.90–32.31), and AUC of 0.89. Subgroup analyses revealed a better performance in patients with ulcerative colitis (AUC, 0.93) than Crohn’s disease (AUC, 0.84). Consistent upregulation of miR-21, miR-16, and miR-192 in blood with a high-moderate diagnostic accuracy was found in at least two studies.ConclusionsThese findings suggest miRNAs are credible diagnostic biomarkers in IBD.

Mucosal Gene Transcript Signatures in Treatment Naïve Inflammatory Bowel Disease: A Comparative Analysis of Disease to Symptomatic and Healthy Controls in the European IBD-Character Cohort.

BackgroundStudies of the mucosal transcriptomic landscape have given new insight into the pathogenesis of inflammatory bowel disease (IBD). Recently, the predictive biomarker potential of gene expression signatures has been explored. To further investigate the mucosal gene expression in IBD, we recruited a cohort of treatment naïve patients and compared them to both symptomatic and healthy controls.MethodsAltogether, 323 subjects were included: Crohn’s disease (N = 75), ulcerative colitis (N = 87) and IBD unclassified (N = 3). Additionally, there were two control groups: symptomatic controls (N = 131) and healthy controls (N = 27). Mucosal biopsies were collected during ileocolonoscopy and gene expression in inflamed and non-inflamed mucosa was explored. Gene expression profiling was performed using Agilent G3 Human Gene Expression 860K v3 One-Color microarray. We recorded information about treatment escalation to anti-TNF agents or surgery, and anti-TNF response, to explore predictive opportunities of the mucosal transcriptome.ResultsGene expression profiles in symptomatic controls in whom IBD had been excluded resembled that of IBD patients and diverged from that of healthy controls. In non-inflamed Crohn’s disease and ulcerative colitis, gene set enrichment analysis revealed dysregulation of pathways involved in basic cellular biological processes. Mitochondria-associated pathways were dysregulated both in non-inflamed and inflamed Crohn’s disease and ulcerative colitis (>2.6 normalized enrichment scores <−1.8). Gene expression signatures of Crohn’s disease and ulcerative colitis did not predict time for treatment escalation (p = 0.175). No significant association was found between gene expression signatures and anti-TNF response.ConclusionNon-inflamed samples are probably superior to inflamed samples when exploring gene expression signatures in IBD and might reveal underlying mechanisms central for disease initiation. The gene expression signatures of the control groups were related to if they were symptomatic or not, which may have important implications for future study designs.