Forms Of Inflammatory Bowel Disease Research Articles

Ulcerative colitis: functional analysis of the in-depth proteome

BackgroundUlcerative colitis (UC) is one major form of inflammatory bowel disease. The cause and the pathophysiology of the disease are not fully understood and we therefor aim in this study to identify important pathophysiological features in UC from proteomics data.MethodsColon mucosa biopsies from inflamed tissue of untreated UC patients at diagnosis and from healthy controls were obtained during colonoscopy. Quantitative protein data was acquired by bottom-up proteomics and furthermore processed with MaxQuant. The quantitative proteome data was analyzed with Perseus and enrichment data was analyzed by ClueGO for Cytoscape.ResultsThe generated proteome dataset is to-date the deepest from colon mucosa biopsies with 8562 identified proteins whereof 6818 were quantified in > 70% of the samples. We report abundance differences between UC and healthy controls and the respective p values for all quantified proteins in the supporting information. From this data set enrichment analysis revealed decreased protein abundances in UC for metallothioneins, PPAR-inducible proteins, fibrillar collagens and proteins involved in bile acid transport as well as metabolic functions of nutrients, energy, steroids, xenobiotics and carbonate. On the other hand increased abundances were enriched in immune response and protein processing in the endoplasmic reticulum, e.g. unfolded protein response and signal peptidase complex proteins.ConclusionsThis explorative study describes the most affected functions in UC tissue. Our results complemented previous findings substantially. Decreased abundances of signal peptidase complex proteins in UC are a new discovery.

Redox modulatory protective effects of ω-3 fatty acids rich fish oil against experimental colitis

Ulcerative colitis (UC), a form of inflammatory bowel disease (IBD), is an immune-modulated disorder characterized by chronic and recurring inflammatory episodes. Oxidative stress and COX pathway of prostaglandin (PG) biosynthesis are indispensable to pathogenesis of UC. Any imbalance between PGs can compromise the mucosal homeostasis, leading to mucosal damage and chronic inflammation. However, blocking these PGs using classical Cox inhibitors such as non-steroidal anti-inflammatory drugs (NSAIDs) can instead aggravate signs of IBD. Therefore, realizing the need for safer and well tolerable alterative treatment approaches, currently, we evaluated the efficacy of n-3 fatty acids rich fish oil (FO) in the resolution of UC. Using a dextran sodium sulfate (DSS) model of experimental colitis, we have demonstrated that supplementation of FO containing 180 mg EPA and 120 mg DHA for 1 month relieved the signs (diarrhea, bloody stools, weight loss) of colitis-associated inflammation. To understand the biophysical changes associated with FO mediated inflammatory regulation, impedance measurement and Fourier transform infrared spectroscopy (FTIR) were done. These changes were also correlated with oxidative stress through markers such as GST, glutathione peroxidase (GPx), LPO, catalase, protein carbonyl content, GR, etc. in colonic mucosa. The modulation of COX mediated pathways in UC-associated inflammation was observed by protein expressions of various pro-inflammatory cytokines such as TNF-α and enzymes of PG synthesis such as COX-2, PGES, TXAS, and anti-inflammatory PGDS. Refuting the earlier reports that suggested the contradictory effects of FO, in the current study, we evidently demonstrated that the protective effects of FO are mediated through molecular mechanisms involving the redox-regulation of metabolism of key lipid metabolites.

Identifying Patients With Inflammatory Bowel Diseases in an Administrative Health Claims Database: Do Algorithms Generate Similar Findings?

Application of selective algorithms to administrative health claims databases allows detection of specific patients and disease or treatment outcomes. This study identified and applied different algorithms to a single data set to compare the numbers of patients with different inflammatory bowel disease classifications identified by each algorithm. A literature review was performed to identify algorithms developed to define inflammatory bowel disease patients, including ulcerative colitis, Crohn’s disease, and inflammatory bowel disease unspecified in routinely collected administrative claims databases. Based on the study population, validation methods, and results, selected algorithms were applied to the Optum Clinformatics® Data Mart database from June 2000 to March 2017. The patient cohorts identified by each algorithm were compared. Three different algorithms were identified from literature review and selected for comparison (A, B, and C). Each identified different numbers of patients with any form of inflammatory bowel disease (323 833; 246 953, and 171 537 patients, respectively). The proportions of patients with ulcerative colitis, Crohn’s disease, and inflammatory bowel disease unspecified were 32.0% to 47.5%, 38.6% to 43.8%, and 8.7% to 26.6% of the total population with inflammatory bowel disease, respectively, depending on the algorithm applied. Only 5.1% of patients with inflammatory bowel disease unspecified were identified by all 3 algorithms. Algorithm C identified the smallest cohort for each disease category except inflammatory bowel disease unspecified. This study is the first to compare numbers of inflammatory bowel disease patients identified by different algorithms from a single database. The differences between results highlight the need for validation of algorithms to accurately identify inflammatory bowel disease patients.

Multiple Cytokine Profiling: A New Model to Predict Response to Tumor Necrosis Factor Antagonists in Ulcerative Colitis Patients.

Ulcerative colitis (UC) is a form of inflammatory bowel disease, and antibodies against tumor necrosis factor (anti-TNF) are used for treatment. Many patients are refractory or lose response to anti-TNF, and predicting response would be an extremely valuable clinical tool. Unlike most biomarkers, cytokines directly mediate inflammation, and their measurement may predict the likelihood of response or no response. Serum samples were obtained from 49 UC patients before infliximab infusions, and levels of 17 cytokines were measured using a multiplex assay. The Fisher linear discriminant analysis (FLDA) was applied to the cytokine values to predict which patients would respond to infliximab. "Response" was defined as clinical remission after the third infusion, and "no response" was defined as lack of remission after the third infusion. The Fisher linear discriminant analysis model identified a subset of seven predictor cytokines: TNF-α, IL-12, IL-8, IL-2, IL-5, IL1-β, and IFN-γ. The obtained canonical coefficients enabled to calculate discriminant scores as linear combinations of the cytokines; model classified thepatients as responders and nonresponders with a sensitivity of 84.2% and a specificity of 93.3%. Overall, the yield of the FLDA model was 89.8% of the total 49 patients. An unbiased, statistically derived, predictive model based on measurement of serum cytokines before therapy may predict a positive or negative outcome from the administration of anti-TNF to UC patients. Because accurately measuring cytokines is simple and inexpensive, the model may be a valuable new tool to complement other laboratory parameters used in the management of IBD patients.

Suppression of miR-21 and miR-155 of macrophage by cinnamaldehyde ameliorates ulcerative colitis

Ulcerative colitis (UC) is a major form of inflammatory bowel disease which involved mucosal immune dysfunction. Cinnamaldehyde (CA) is major active compound from cinnamon, a useful traditional medicine in Asia which shows superior antibacterial and anti-inflammatory activity. In this study, we investigated the effects of CA on UC both in vivo and in vitro. We showed that CA attenuated the symptoms of DSS-induced colitis, including loss of body weights, disease activity index (DAI), shortening of the colon lengths and infiltration of inflammatory cells. Moreover, CA decreased the pro-inflammatory cytokines and NLRP3 inflammasome, miR-21 and miR-155 in colon tissues, in addition, the percentage of macrophages was reduced based on the surface marker F4/80 and IL-10 secretion in CA-treated group, suggesting that the CA ameliorate the UC via activation of macrophage. Herein, the effects of CA on macrophage cells were examined in vitro. We found that CA reduced the level of proinflammatory cytokines, such as TNF-α, IL-1β, IL-6, in the activation of RAW264.7, human macrophage-like cells U937, and primary peritoneal macrophages. Furthermore, the suppression of NLRP3 inflammasome, miR-21 and miR-155 was also found in CA-treated LPS-stimulated RAW264.7 cells. CA also reduced the production of reactive oxygen species, the phosphorylation of AKT, mTOR and COX2 protein level in the RAW264.7. Meanwhile, data revealed that transferred miR-21 or miR-155 inhibitor suppressed levels of IL-1β and IL-6, whereas miR-21 or miR-155 mimics increased expressions of these, and CA suppressed these expressions. Our results indicate that CA could ameliorate DSS-induced colitis through inhibition of NLRP3 inflammasome activation and miR-21 and miR-155 levels in colons and macrophage, suggesting that CA might be a potentially effective drug for UC.

Comparative analysis of Faecalibacterium prausnitzii genomes shows a high level of genome plasticity and warrants separation into new species-level taxa

BackgroundFaecalibacterium prausnitzii is a ubiquitous member of the human gut microbiome, constituting up to 15% of the total bacteria in the human gut. Substantial evidence connects decreased levels of F. prausnitzii with the onset and progression of certain forms of inflammatory bowel disease, which has been attributed to its anti-inflammatory potential. Two phylogroups of F. prausnitzii have been identified, with a decrease in phylogroup I being a more sensitive marker of intestinal inflammation. Much of the genomic and physiological data available to date was collected using phylogroup II strains. Little analysis of F. prausnitzii genomes has been performed so far and genetic differences between phylogroups I and II are poorly understood.ResultsIn this study we sequenced 11 additional F. prausnitzii genomes and performed comparative genomics to investigate intraspecies diversity, functional gene complement and the mobilome of 31 high-quality draft and complete genomes. We reveal a very low level of average nucleotide identity among F. prausnitzii genomes and a high level of genome plasticity. Two genomogroups can be separated based on differences in functional gene complement, albeit that this division does not fully agree with separation based on conserved gene phylogeny, highlighting the importance of horizontal gene transfer in shaping F. prausnitzii genomes. The difference between the two genomogroups is mainly in the complement of genes associated with catabolism of carbohydrates (such as a predicted sialidase gene in genomogroup I) and amino acids, as well as defense mechanisms.ConclusionsBased on the combination of ANI of genomic sequences, phylogenetic analysis of core proteomes and functional differences we propose to separate the species F. prausnitzii into two new species level taxa: F. prausnitzii sensu stricto (neotype strain A2–165T = DSM 17677T = JCM 31915T) and F. moorei sp. nov. (type strain ATCC 27768T = NCIMB 13872T).

Interleukin-10 Signaling in Hematopoietic Stem and Progenitor Cells Maintains Stem Cell Function and Regulates Inflammation-Induced Myeloid Cell Output

Inflammation shapes the immune response to foreign antigens by influencing effector immune function and is dysregulated in many forms of inflammatory bowel disease and other auto-immune conditions. It is increasingly recognized that inflammation also impacts hematopoietic stem and progenitor cell (HSPC) function. Interleukin-10 (IL-10) is an anti-inflammatory cytokine that regulates inappropriate immune responses by suppressing inflammatory programs in mature immune cells. However, the direct role of IL-10 in modulating HSPC function and in suppressing hematopoietic cell output in the context of inflammation has not previously been demonstrated.IL10Rß-deficient (Il10rb-/-) mice have an increased inflammatory cytokine profile and develop overt colitis symptoms between 4-6 months of age. We observed an expansion in the relative frequency of immunophenotypically-defined HSPCs (LSK+: Lineage-c-Kit+Sca-1+) and frequency and absolute numbers of granulocyte-monocyte progenitors (GMPs: Lineage-c-Kit+CD34+FcgRII/III+) in pre-colitic Il10rb-/- mice relative to wildtype mice (WT) (see table). Increased GMPs correlated with increased myeloid and reduced lymphoid frequency and number in the bone marrow (BM). Consistent with the expansion in the Il10rb-/-BM HSPC compartment, transplantation of a 1:1 ratio of Il10rb-/-: WT whole BM also resulted in a 2-fold higher short-term reconstitution of Il10rb-/- CD45+ cells in peripheral blood relative to WT cells. Moreover, in vitro we observed suppressed expansion of WT but not Il10rb-/- lineage- cells after IL-10 treatment, suggesting that IL10 suppresses HSPC proliferation. To determine if IL-10 signaling affects the function of hematopoietic stem cells (HSCs), we compared an equal number of purified HSCs (LSK+CD150+CD48-) from Il10rb-/- and WT mice in repopulation assays. Il10rb-/- HSCs demonstrated a 6-fold (P<0.01) reduced lineage reconstitution in peripheral blood relative to WT HSCs in competitive transplantation, thus demonstrating that, while Il10rb-/- mice have an increased pool of committed progenitor cells, HSC function is reduced relative to WT mice. To confirm direct IL-10 signaling in HSPC populations, purified LSK+ and myeloid progenitors (LK+Sca-1-) were exposed to IL-10. Significant STAT3 phosphorylation was observed in HSPCs and myeloid progenitors, with the highest proportion of phosphorylated cells in the GMP fraction. These data suggest that IL-10 directly acts on multiple stem and progenitor populations and that myeloid progenitors robustly respond to IL-10. To determine if the alterations in Il10rb-/- BM are cell intrinsic or a product of underlying inflammation in the absence of IL-10 signaling in vivo, we generated mixed whole BM WT: Il10rb-/- chimeric mice and induced inflammation through E. coli lipopolysaccharide (LPS) injection. LPS exposure led to a 1.5-fold larger depletion of Il10rb-/- GMPs compared to WT GMPs in mixed chimeric mice 24 hours post-injection relative to vehicle treatment. These data suggest a protective role for IL-10 on progenitors during an acute inflammatory insult. Upon transplantation of BM from primary LPS-treated mixed chimeric mice, a short-term increase in peripheral blood myeloid cells was observed when compared to transplanted BM from vehicle-treated mice. Notably, the enhanced peripheral blood myeloid output from LPS-treated donor BM was skewed 2-fold in favor of Il10rb-/- cells and BM myeloid colony-forming-cells were skewed 4-fold in favor of Il10rb-/- cells relative to WT cells. These data demonstrate that IL-10 attenuates inflammation-induced myeloid output upon BM transplantation.Taken together, this study reveals a previously unrecognized role for IL-10 in suppressing progenitor cell expansion and promoting HSC function at steady state while mitigating myeloid lineage skewing after an inflammatory insult. [Display omitted] DisclosuresWilliams:Bluebird Bio: Research Funding.

Specific Features of Vascular Endothelium in Patients with Severe Forms of Inflammatory Bowel Diseases

Aim.In this work, we set out to assess the specific features of vascular endothelium in patients suffering from severe (pan-ulcerative) and complicated forms of ulcerative colitis (UC) and Crohn’s disease (CD).Materials and methods.The research sample consisted in 65 patients with UC (n = 45) and CD (n = 20), who were either in the acute phase or had been diagnosed with such conditions for the first time. For assessing endothelial dysfunction (ED), we used such markers as levels of vascular endothelial growth factor (VEGF) in the blood serum and the number of plasma desquamated endotheliocytes (DEC).Results.The indicators of vascular endothelial dysfunction are shown to be significantly higher in patients with severe, pan-ulcerative forms of inflammatory bowel diseases (IBD). A statistically significant relationship has been established between ED indicators and the severity and localisation of the pathological process, the extra-intestinal manifestations and complications of the disease. A dependence of the endothelial dysfunction indicators on the UC endoscopic activity has been determined. A positive correlation between the levels of VEGF and DEC and the markers of systemic inflammation (ESR, CRP and fibrinogen) has been revealed.Conclusion. It is determined that, in IBD patients, endothelium dysfunction is manifested in an increase in the VEGF and DEC levels. The indicators of endothelial dysfunction are shown to be directly correlated with the serum markers of systemic inflammation. Therefore, VEGF and DEC values can be used not only as criteria for assessing the intensity of IBD, but also as predictors of a complicated disease course.

Refolding, Characterization, and Preliminary X-ray Crystallographic Studies on the Campylobacter concisus Plasmid-Encoded Secreted Protein Csep1p Associated with Crohn’s Disease

Colonization of Campylobacter concisus in the gastrointestinal tract can lead to the development of inflammatory bowel disease (IBD). Plasmid-encoded C. concisus-secreted protein 1 (Csep1p) was recently identified as a putative pathogenicity marker associated with active Crohn’s disease, a clinical form of IBD. Csep1p shows no significant full-length sequence similarity to proteins of known structure, and its role in pathogenesis is not yet known. This study reports a method for extraction of recombinantly expressed Csep1p from Escherichia coli inclusion bodies, refolding, and purification to produce crystallizable protein. Purified recombinant Csep1p behaved as a monomer in solution. Crystals of Csep1p were grown by the hanging drop vapour diffusion method, using polyethylene glycol (PEG) 4000 as the precipitating agent. A complete data set has been collected to 1.4 Å resolution, using cryocooling conditions and synchrotron radiation. The crystals belong to space group P62 or P64, with unit cell parameters a = b = 85.8, c = 55.2 Å, α = β = 90, and γ = 120°. The asymmetric unit appears to contain one subunit, corresponding to a packing density of 2.47 Å3 Da−1.

Abstract PR06: Drugging the human microbiome for combination with tumor immunotherapy

Abstract The human gut microbiome is a diverse, dynamic, and complex ecosystem that modulates host processes including metabolism, inflammation, and cellular and humoral immune responses. Recent studies have suggested that the microbiome may also influence the development of certain cancers such as colorectal cancer, and equally importantly, tumor response to systemic therapy, especially immunotherapy. Multiple groups are exploring the therapeutic utility of the microbiome to enhance clinical response through the use of defined oral therapeutics comprising living commensal bacteria, which would represent a new therapeutic modality. Exploiting the microbiome for therapeutic benefit is not without its challenges due to the heterogeneity of the gut microbiota across healthy donors and patients. In addition, many aspects of conventional small molecule and biologics drug discovery and development do not apply to this novel class of living drugs. We present an approach that leverages the concept of “reverse translation,” using genomic and immunologic characterization of patient samples from interventional studies to define and better understand the organisms and mechanisms that contribute to response or non-response to immunotherapy. We are investigating the relationship between the composition of the gut microbiome prior to therapy and the antitumor response in patients receiving checkpoint inhibitors (CPI), as well as how CPI treatment modulates the microbiome in both responders and nonresponders. Fecal and blood samples are collected before and during therapy from cancer patients who receive approved CPI; tumor types include renal, bladder, and NSCLC. Whole metagenomic shotgun sequencing of patient microbiomes is used to identify higher order (e.g., order- and family-level) “microbial signatures” that associate with response to CPI treatment. We then utilize proprietary algorithms that enable species- and strain-level resolution of microbial signatures. In addition, global and targeted metabolomics are used to identify functional pathways associated with outcome, and these pathways can be linked to species and strains identified by genomic analysis. Our discovery strategy iterates computational analyses and machine learning approaches with empirical in vitro and ex vivo screening of strains and consortia to inform selection and drive drug design. Data from such a comprehensive approach is invaluable for designing compositions of bacteria that form “functional ecological networks” that can impact response to CPI therapy. Finally, our microbial library of &amp;gt;14,000 isolates from healthy human subjects captures the phylogenetic diversity and functional breadth of the gastrointestinal microbiome, and provides a robust platform to build unique compositions. Such compositions, when tested in syngeneic tumor models in germ-free mice, can provide a preliminary readout of the contributions of members of the consortia and enable candidate identification. We present examples of reverse translation in patients with recurrent Clostridium difficile infection and ulcerative colitis, a form of inflammatory bowel disease, that have led to the translation of three drugs that are currently in clinical trials. This roadmap provides insight into how similar drugs can be discovered and developed in the setting of immunotherapy to augment the efficacy of CPIs by altering the cancer-immune set point. This abstract is also being presented as Poster A06. Citation Format: David N. Cook, Jonathan Peled, Marcel van den Brink, Lata Jayaraman. Drugging the human microbiome for combination with tumor immunotherapy [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2017 Oct 1-4; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2018;6(9 Suppl):Abstract nr PR06.

Modelling the Inflammatory Bowel Disease Specialist Nurse Workforce Standards by Determination of Optimum Caseloads in the UK.

Crohn's disease and ulcerative colitis are the most common forms of inflammatory bowel disease [IBD], affecting 1 in 250 of the population in the UK. It is accepted that access to a specialist nurse service improves patient experience and outcome. National Standards for the Healthcare of People Who Have Inflammatory Bowel Disease [2013] defined the number of nurse specialists required at 1.5 full time equivalent per 250000 population. The aim of this study was to determine if these standards were being met and to publish a new, robust, validated standard optimising the UK nursing workforce model. Existing national data and specific workload and service data were collected from 164 IBD specialist nurses who completed a questionnaire designed to collect information on activity and complexity of work both done and undone. Data were received from all of the UK; 36% of respondents were specialist nurses in the field for 3 years or less. A higher caseload than the recommended level was reported by 63% of respondents. Unpaid overtime was regularly carried out by 84% of respondents. The IBD specialist nurse was involved in all areas of the patient pathway. Areas of work left undone were psychological interventions, prescribing medicines and physical assessments. Compared with other specialties, IBD specialist nurses are less experienced. It is recommended that the current standard be increased to 2.5 full time equivalent specialist nurses per 250000 patients, to mitigate the increasing complexity of the role, care and the 'rookie' factor.

Genistein induces macrophage polarization and systemic cytokine to ameliorate experimental colitis.

Mucosal changes in Crohn’s disease (CD) and ulcerative colitis (UC), two major forms of inflammatory bowel disease (IBD), are characterized by a prominent infiltration of inflammatory cells including lymphocytes, macrophages, T cells and neutrophils. The precise etiology of IBD is unknown but it involves a complex interplay of factors associated with the immune system, environment, host genotype and enteric commensal bacteria. As there is no known safe cure for IBD, natural alternative therapeutic options without side effects are urgently needed. To this end, Soy-based foods, which have been eaten for centuries in Asian countries, have potential benefits, including lowering the incidence of coronary heart disease, atherosclerosis, type-2 diabetes, allergic response, and autoimmune diseases. This study describes the effect of Soy isoflavons 4', 5, 7 Trihydroxyisoflavone (genistein) on dextran sodium sulphate (DSS) induced experimental colitis. The extent and severity of disease was analyzed through body weight, histopathological analysis, cellular immune response, systemic cytokine levels, and inflammation score using a disease activity index. Genistein treatment significantly attenuated DSS-induced colitis severity and resulted in increase in body weight, colon length and reduction in inflammation score. Genistein also skews M1 macrophages towards the M2 phenotype. Further, gen also reduced the systemic cytokine levels as compared to vehicle control. This serves as the first detailed study towards natural soya based product that shows the polarization of M1 towards M2 macrophages, and reduction of systemic cytokine in part to attenuate the colitis symptoms. Thus, our work demonstrates that genistein, a soya compound, may be useful for the treatment of IBD.

Cancer-predicting transcriptomic and epigenetic signatures revealed for ulcerative colitis in patient-derived epithelial organoids

Ulcerative colitis (UC) is a prevalent form of inflammatory bowel disease (IBD) whose pathogenic mechanisms remain unclear. Elucidating these mechanisms is important to reduce UC symptoms and to prevent UC progression into colitis-associated colon cancer (CAC). Our goal was to develop and validate faithful, human-derived, UC models and analyze them at histologic, transcriptomic and epigenetic levels to allow mechanistic studies of UC and CAC pathogenesis.We generated patient-derived primary-organoid cultures from UC and non-IBD colonic epithelium. We phenotyped them histologically and used next-generation-sequencing approaches to profile whole transcriptomes and epigenomes of organoids and primary tissues.Tissue organization and expression of mucin 2 (MUC2) and lysozyme (LYZ) demonstrated histologic faithfulness of organoids to healthy and diseased colonic epithelium. Transcriptomic analyses showed increased expression of inflammatory pathways in UC patient-derived organoids and tissues. Profiling for active enhancers using the H3K27ac histone modification revealed UC-derived organoid enrichment for pathways indicative of gastrointestinal cancer, including S100 calcium-binding protein P (S100P), and revealed novel markers for GI cancer, including both LYZ and neuropeptide S receptor 1 (NPSR1). Immunolocalization showed increased levels of LYZ, S100P, and NPSR1 proteins in UC and CAC.In conclusion, primary colonic organoid cultures from UC and non-IBD patients can be established that faithfully represent diseased or normal colonic states. These models reveal precancerous molecular pathways that are already activated in UC. The findings demonstrate the suitability of primary organoids for dissecting UC and CAC pathogenic mechanisms and suggest new targets for therapeutic intervention.

Therapeutic effects of Lactobacillus paracasei subsp. paracasei NTU 101 powder on dextran sulfate sodium-induced colitis in mice

Ulcerative colitis (UC) is a form of inflammatory bowel disease (IBD) whose exact cause is still unclear. Disruption of the intestinal microflora is considered one of the main causes of the disease. Lactobacillus paracasei subsp. paracasei NTU 101 (NTU 101) is a multifunctional strain that has been shown in previous studies to possess anti-inflammatory properties and to exert a modulatory effect on intestinal bacteria associated with certain pathogenic mechanisms of IBD. In the current study, we investigated the effects of NTU 101 on dextran sulfate sodium (DSS)-induced colitis in a mouse model. Colitis was induced in male C57BL/6 mice (total number n = 60) via dissolved DSS in drinking water on days 15–21 of the experiment. The effects of continuous 25 d feeding (days 0–25) of either a half or a full dose [2.3 × 109 colony-forming units (CFU)/kg body weight (BW)/d and 4.5 × 109 CFU/kg BW/d, respectively] of NTU 101 was evaluated. Lactobacillus rhamnosus BCRC 16000 (BCRC 16000) and L. paracasei subsp. paracasei BCRC 14023 (BCRC 14023) strains were given to control groups. The results indicated that NTU 101 powder improved anti-oxidant capacity, reduced pro-inflammatory cytokine levels, increased anti-inflammatory cytokine levels, and slightly ameliorated body weight loss in DSS-treated mice during the final days of the study. This indicated that NTU 101 powder can relieve the clinical symptoms of DSS-induced colitis in mice.

Protein Kinase 2 (CK2) is Required for CD4+ T Cell Function in the Pathogenesis of Colitis

Abstract Crohn’s disease (CD) is one of the major forms of inflammatory bowel disease (IBD), characterized by chronic inflammation of the gastrointestinal tract. CD is associated with aberrant Th1 and Th17 responses accompanied by high levels of IFN-γ and IL-17, respectively. Protein kinase CK2 is a highly conserved serine-threonine kinase that is involved in several signal transduction pathways, including the JAK/STAT, NF-κB, PI3K/Akt/mTOR and Wnt pathways. Inhibition of CK2 kinase activity by small molecule inhibitor (CX-4945) or genetic deletion of CK2α in CD4+ T cells shifts the balance between Th17 and Tregs. However, the function of CK2 in the pathogenesis of colitis is unknown. Here, we demonstrate that CD4+ T cells from CK2αfl/fl dLck-Cre mice failed to induce wasting disease and intestinal inflammation, accompanied by lower levels of inflammatory cytokines and chemokines in the serum, and decreased IL-17A+, IFN-γ+ and IL-17A+/IFN-γ+ CD4+ T cells in the spleen and colon. Furthermore, CK2α controls CD4+ T cell accumulation in the spleen and colon through a cell-intrinsic mechanism. Thus, our data suggest that CK2α contributes to the pathogenesis of colitis, and that inhibition of CK2 kinase activity may be a novel therapeutic treatment for patients with CD.

Interleukin-7 receptor pathway controls human T cell homing to the gut and predicts response to anti-TNFα therapy in IBD

Abstract Background and aims Signaling networks perpetuating chronic gastrointestinal inflammation in Crohn’s disease (CD) and ulcerative colitis (UC), the two main forms of inflammatory bowel diseases (IBD), remain unclear in man. Methods and results According to an analysis of nearly 500 patients with IBD and 100 controls, we report here that key transcripts of the IL-7 receptor (IL-7R) pathway accumulate in inflamed colon tissues of severe IBD patients not responding to either immunosuppressive/corticosteroids or anti-TNFα therapies. High expression of both IL-7R and IL-7R signaling signature transcripts in the colon before treatment are strongly associated with nonresponsiveness to anti-TNFα therapy. While in mice IL-7 is known to play a role in systemic inflammation, we found that in humans IL-7 also controls α4/β7 integrin expression and imprints gut-homing specificity on T cells. IL-7 receptor blockade in vivo reduced human T cell homing to the gut and colonic inflammation in humanized mouse models, and decreased production of IFNγ by colon explants from UC patients grown ex vivo. Conclusion Our findings suggest that the failure of current treatments for IBD, including anti-TNF, may occur at least in part by dysregulated IL-7R signaling and points to the IL-7R as a relevant novel therapeutic target and biomarker to fill an unmet need in clinical IBD detection and treatment.

Decreased risk for microscopic colitis and inflammatory bowel disease among patients with reflux disease.

Previous studies have found an increased risk for microscopic colitis (MC) associated with proton pump inhibitors. In patients with ulcerative colitis (UC) or Crohn's disease (CD), proton pump inhibitors have been associated with an increased risk for IBD flares and impaired outcomes. The aim of this study was to test the epidemiological associations between gastro-oesophageal reflux disease (GERD) and MC, UC or CD in a large database. The Miraca Life Sciences Database is a national electronic repository of histopathological records of patients distributed throughout the entire USA. A case-control study evaluated whether the presence of Barrett's metaplasia, erosive oesophagitis on endoscopy or histological signs of reflux oesophagitis, clinical diagnosis of GERD or any GERD type affected the occurrence of MC, UC or CD among 228506 subjects undergoing bidirectional endoscopy. Multivariate logistic regression analyses were used to calculate ORs and their 95% CI for the risk of MC, UC or CD associated with various types of GERD and were adjusted for age, sex and presence of Helicobacter pylori. The analysis revealed an inverse relationship between GERD and different types of inflammatory bowel disease. The inverse relationships applied similarly to MC (mean=0.62, 95% CI: 0.58-0.66), UC (mean=0.89, 95% CI: 0.81-0.97) and CD (mean=0.76, 95% CI: 0.69-0.85). It also applied to different forms of GERD, with a trend towards more pronounced inverse relationships associated with Barrett's metaplasia or erosive oesophagitis than clinical diagnosis of GERD. Gastro-oesophageal reflux disease is inversely associated with all forms of inflammatory bowel disease, such as MC, UC, or CD.

Nerolidol a Natural Aliphatic Sesquiterpene Mitigates Inflammation in DSS‐induced Colitis

IntroductionCrohn's disease and ulcerative colitis are the two main forms of inflammatory bowel disease (IBD) which affect both small and large intestine. The aetiology of this disease remains unclear but seems to be multifactorial in nature. The current therapy for IBD relies on the use of sulfasalazine, corticosteroids, azatriopine and anti‐TNFa antibody as a mainstream therapy. However, the adverse effect associated with these drugs over prolonged treatment period limit their use. Therefore, unconventional treatment using dietary supplements rich in phytochemicals is increasing in popularity because of less adverse effect along with time tested safety and acceptability. Phytochemicals (a group of secondary plant metabolites) richly present in plants and plant based food sources. Numerous line of evidences implicate that the consumption phytochemical rich diet is beneficial in lowering the risk of several diseases. The anti‐inflammatory effect of many phytochemicals are partly mediated by modulating nuclear factor kappa B (NFκB) directly or indirectly through activating peroxisome proliferator activated receptor gamma along with its proven antioxidant properties. These mounting evidences suggest that the prospect of phytochemical mediated effect is most likely to encompass more than the well‐known antioxidant activity. Nerolidol (NRD) is one such phytochemical found abundantly in many edible plants appears to be having anti‐inflammatory properties.AimTherefore, the main aim of this study was to investigate the role of NRD for its anti‐inflammatory property using mice model of colonic inflammation.MethodologyC57Bl/6 black mice were orally administered with 3% dextran sodium sulfate (DSS) in drinking water for 7 days to induce colitis. The treatment group received NRD using oral gavage at various concentrations (50–150 mg/kg body wt) and compared with control and DSS along with a positive control through clinically used drug (sulfasalazine) group. The disease activity index (DAI) was established throughout the treatment period. After 7 days, the colon was excised and the length was measured processed for histology. Changes in myeloperoxidase (MPO) activity and pro‐inflammatory cytokines were estimated using ELISA and RTPCR. Cox‐2, and iNOS protein expression was carried out using Western blot.ResultsNRD significantly (p&lt;0.01) decreased DAI dose dependently. DSS administration significantly (p&lt;0.01) reduced colon length however, NRD reversed colon length changes dose dependently. MPO activity was also significantly (p&lt;0.01) inhibited by NRD dose dependently. Histological scoring for colonic crypt damage and inflammation was significantly improved upon NRD treatment. NRD also inhibited the pro‐inflammatory cytokine (IL‐1β, IL‐6 and TNF‐α) response significantly both at protein and mRNA level. COX‐2 and iNOS protein expression was also significantly inhibited by NRD.ConclusionNRD appears to attenuate aberrant inflammatory response observed in DSS‐induced colitis by inhibiting pro‐inflammatory mediators.Support or Funding InformationStart‐Up funds from UAE UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Loss of Sprouty2 enhances IL‐33 expression and protects against experimental colitis.

OBJECTIVESprouty2 (Spry2) is an intracellular modulator of MAPK signaling. It is present in the intestinal tract, but its function there is unknown. Aberrant MAPK signaling is associated with altered cytokine production in disorders such as inflammatory bowel disease, raising the possibility that Spry2 may affect the response to colitis. In particular, MAPK regulates IL‐33 expression in response to bacterial infection, and IL‐33 has been shown to be protective in some forms of colitis. Furthermore, both MAPK and IL‐33 regulate secretory cell differentiation. Here we tested the hypothesis that Spry2 control of MAPK signaling alters cytokine production and/or secretory cell development, thus altering the intestinal response to injury.METHODSMice that lack Spry2 in the intestinal epithelium (VillinCre;Spry2flox/flox, hereafter Spry2KOIE) and littermate controls were given 3% dextran sodium sulfate (DSS) in drinking water for 4 days followed by 3 days without DSS, and colitis severity was assessed. Cytokine levels and epithelial marker expression were determined by qPCR in challenged and unchallenged mice. GSK3beta phosphorylation was determined by western blot. In vitro, IEC6 and YAMC intestinal epithelial cells were treated with GSK3beta and MAPK inhibitors and cytokine expression was assessed.RESULTSColonic Spry2 levels were reduced in DSS colitis by 59% (p&lt;0.05). Spry2KOIE mice subjected to DSS had lower levels of pro‐inflammatory cytokines versus wildtype littermates, and were protected from histological damage (injury score reduced by 30%, p=0.037), weight loss, and colon shortening. Unchallenged Spry2KOIE mice displayed elevated GSK3beta phosphorylation on the inactivating Ser9 site in epithelial cells (88% increase, p&lt;0.01). These mice also had increased IL‐33 expression (80% increase, p&lt;0.05) and more DCLK1+ tuft cells in the colon (2.2‐fold increase, p&lt;0.01). In vitro, inactivation of GSK3beta in both IEC6 and YAMC cells mimicked the increased IL‐33 observed in vivo; this effect was attenuated by ERK or p38 MAPK inhibitors. Expression of the epithelial‐derived cytokine CXCL2 was unaffected by GSK3beta inhibition, suggesting an IL‐33 specific response.CONCLUSIONColonic epithelial Spry2 is downregulated in response to injury and inflammation. This may represent an integration point of environmentally‐driven MAPK signals that leads to altered Wnt signaling, elevated IL‐33 expression, and promotion of intestinal tuft cell development as a protective or regenerative mechanism.Support or Funding InformationNIH R01DK095004This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Tracking inflammation in the colon

Inflammation The therapeutic options for ulcerative colitis, a form of inflammatory bowel disease, are limited. Lyons et al. tracked gene expression, protein levels, and protein phosphorylation in individual animals in a mouse model of colitis. Computational analysis of the data sets identified