bluuebunny/crossref_metadata_2025_split huggingface dataset:
A207 RIPK2 AND AMPK AS EMERGING THERAPEUTIC TARGETS FOR INFLAMMATORY BOWEL DISEASE
Authors: Guo Y , Dieleman L A , Dooky H , Joshi H , Wine E , Baksh S 
Published: February 2020

Abstract

Background
Persistent inflammation can trigger altered epigenetic, inflammation and bioenergetics states. Inflammatory bowel disease (IBD) is a heterogeneous disease with an abnormal inflammatory state and subsequent metabolic syndrome disorder. Current IBD therapeutics are directed to inhibit inflammatory pathways, such as inflammation of the epithelial cells in the colonic crypts. We hypothesize that in order to achieve mucosal healing and to keep patients in remission we must (i) inhibit inflammatory mediators and (ii) resolve secondary effects of inflammation such a reset of metabolic dysfunction.

Aims
The aims of this study are to explore correlations between inflammation/metabolic markers and the severity of the disease to uncover emerging new therapeutic players.

Methods
More than one hundred patients were recruited and underwent colonoscopy. In order to explore how biomarkers change with disease and time, all patients had IBD for more than 10 years or less than 5 years. Those diagnosed with cancer, celiac sprue, or diabetes were excluded. The activity of key metabolic markers (such as AMPK) was tracked using phospho-specific antibodies. Immunohistochemistry and immunoblotting were carried out, as described by Gordon et al., PLOSone 2013. All patients were consented under our IBD ethics protocol (Pro00001523 and Pro00077868).

Results
Using intestinal biopsies from non-IBD, UC and CD patients, we explored the expression/activation levels of markers of inflammation (such as obligate NOD2 kinase RIPK2) and metabolism (AMPK) in order to gain insight into correlations with clinical severity of the disease. We confirm that the loss in the activity of AMPK occurs with a gain of activity of RIPK2 that drives the inflammatory phenotype of the gut in patients with long-standing IBD (&gt;10 years). (If inflammation is inhibited in a mouse model of IBD, metabolic reset occurs to regain AMPK and promote mucosal healing). However, RIPK2 remains elevated in patients that are currently on IBD therapeutics.

Conclusions
Therapeutics inhibiting inflammation (RIPK2) and stimulating metabolic (AMPK) drivers of the disease may be a useful combination therapy to completely eliminate inflammation, reset abnormal metabolism and achieve full remission in IBD patients with longstanding disease.

Funding Agencies
None
A212 CHROMOFUNGIN PROTECTS AGAINST DSS-INDUCED COLITIS BY REGULATING P-53 APOPTITIC PATHWAY
Authors: Diarra A , Eissa N , Ghia J 
Published: February 2020

Abstract

Background
Development of ulcerative colitis is associated with epithelial apoptosis mediated by p53-apoptotic pathway through the activation B-cell lymphoma 2 (Bcl-2), Bcl-2 associated-X protein (BAX) and Bcl-2 antagonist/killer-1 (BAK1) proteins. Chromofungin (CHR), a chromogranin-A derived peptide expressing a cell penetrating peptide motif, decreased the severity of colitis via the suppression of mucosal and pro-inflammatory macrophages-related p53-dependent apoptosis.

Aims
We aimed to investigate a) whether the gene profile expression of apoptosis could be extended to other p53-associated genes; b) whether the gene expression of some of the p53-apoptosis marker could be confirmed by protein analysis; and c) whether due to the cell penetrating peptide motif, CHR could enter into peritoneal macrophages.

Methods
UC-related colitis was induced in C57BL/6 mice (7 weeks) by administering dextran sulfate sodium (DSS) (5%, 5 days). Preventive CHR (2.5 mg/kg/day) or vehicle treatment started 1-day before colitis induction and lasted for 5-days. Profiler™ PCR Array was performed to screen a panel of 84 genes representative of the p53 signal pathway in colitic whole mucosa distal colonic samples treated or not with CHR. Western blot analysis was performed to confirm individual protein changes. Naïve macrophages were plated overnight and nonadherent cells were removed the next day. Cells were incubated with rhodamined CHR (4 ul) for 5, 10, 20, 30 min before washing and fixing them, detection was made via confocal microscopy.

Results
In colitic conditions, an up regulation of 26 genes associated to the p53-dependent apoptosis pathway were detected including Apaf1, Bax, Bbc3, Bcl2, Cradd, Fadd, Cul9, Pmaip1, Tnfrsf10b. In vivo CHR treatment decreased significantly the colitis and was associated with a significant downregulation of 19 genes including the 9 aforementioned when compared with biopsies from colitic groups. Compared to untreated groups, colitic mice treated with CHR demonstrated a significant decrease of BAX and BAK protein and the apoptotic ER stress inducer marker, X-Binding Protein 1. A large number of peritoneal macrophages displayed rhodamine within the all intracellular compartment. The presence of the peptide inside the cell can be visible as early as 5 min and the signal gradually increases.

Conclusions
CHR decreases the inflammatory process via the suppression of a large number of p53-related apoptotic proteins. CHR quickly enters the macrophage but the exact mechanism of entrance needs to be further defined. Targeting functional analysis of CHR may lead in the future to novel therapeutics for UC.

Funding Agencies
CCCNSERC
A221 CROHN’S DISEASE POLYGENIC RISK SCORE IS ASSOCIATED WITH FECAL CALPROTECTIN CONCENTRATION IN ASYMPTOMATIC FIRST-DEGREE RELATIVES OF CROHN’S DISEASE PATIENTS
Authors: Lee S , Shestopaloff K , Espin-Garcia O , Turpin W , Raygoza Garay J , Power N , Smith M , Silverberg M , Xu W , Paterson A D , Croitoru K 
Published: February 2020

Abstract

Background
Fecal calprotectin concentration (FC), a measure of gut inflammation is reported to be significantly higher in healthy first-degree relatives (FDR) of Crohn’s disease (CD) patients compared to healthy controls. In contrast, FC in spouses of CD patients was not significantly different from controls, suggesting that a genetic predisposition rather than a shared environmental factor affects FC.

Aims
We investigated the genetic association with FC in healthy FDRs of CD patients. Notably, these subjects are known to be enriched with CD risk alleles.

Methods
We investigated 1455 healthy Caucasian FDRs of CD patients from the GEM Project. Subjects were genotyped by HumanCoreEXOME chip and ImmunoChip platforms and then imputed by the Haplotype Reference Consortium v1.1 panel (Michigan Imputation Server). SNPs with a minor allele frequency&lt;5% were removed. FC was measured using BUHLMANN ELISA kit. Heritability was estimated using a pedigree based SOLAR program and a SNP-based GCTA software. Genome wide association of FC was tested using the GEE framework that accounts for family clusters, age, sex, first 3 genetic principal components and multiplex family status (≥2 FDRs diagnosed with CD). In addition, CD-polygenic risk scores were derived based on summary statistics and imputed SNPs from a recent GWAS by pruning and thresholding (P+T) and LDPred algorithm (PMID:31002795).

Results
Among 1455 subjects, 45.2% were male, median age was 19 years (IQR 13–26), 8.8% were from multiplex families, and median FC was 52 mg/kg (IQR 31–87; 20.8% had FC&gt;100). We estimated the heritability of FC to be 27% (27.1%, standard error=9%, p&lt;0.001 by pedigree approach; 27.9%, SE=12%, p&lt;0.001 by SNP approach). An untargeted GWAS failed to show any significant association with FC (i.e. p&lt;5x10-8). The lowest p value was obtained for rs224631 (p=5x10-7). Strikingly, an increase in CD polygenic risk scores was significantly associated with an increase of FC (p=5.2x10-5 with P+T method).

Conclusions
We demonstrate that FC concentration is a heritable trait in unaffected FDRs of CD patients. Although the association between genetic variants with FC did not reach GWAS significance, CD-polygenic risk score, which incorporates small effect size CD-associated SNPs, was significantly associated with FC concentrationin this cohort. Our results suggest that FC concentration is influenced genetically with contributions from CD-associated SNPs in unaffected FDRs of CD probands. It remains to be determined if the genetic influence to FC concentration is dependent/independent with the future development of CD.
Submitted on behalf of The CCC-GEM Project research team

Funding Agencies
CCCHelmsley Charitable Trust/ Mount Sinai Hospital Fellowship Award
A230 GAPS IN IDENTIFYING FOOD-RELATED QUALITY OF LIFE AND HYPERVIGILANCE IN AT-RISK INDIVIDUALS WITH IBD: REVIEW OF VALIDATED SCREENING TOOLS
Authors: Sasson A N , Targownik L , Vagianos K , Bernstein C N 
Published: February 2020

Abstract

Background
Inflammatory bowel disease (IBD) has a profound impact on psychosocial aspects of food and nutrition, thereby affecting food-related quality of life (QoL). While there is literature on associated prevalence of increased anxiety in individuals with IBD, there is limited data on its contribution to hypervigilance and orthorexia with food intake. Many patients with IBD have strong beliefs about dietary symptom triggers, which can lead to avoidance behaviors and decreased nutrient intake. This can exacerbate malnutrition and disordered eating, as well as increase disease-related stress and negatively impact coping. While there are validated scales evaluating anxiety-related, QoL-related and food-related behaviors in IBD, a combined screening tool to assess this comorbid axis is not well described.

Aims
To conduct a systematic review of existing literature in order to inform practice and facilitate development of an effective food-related hypervigilance and QoL evaluation in IBD patients

Methods
The literature was systematically searched through September 2019, using an electronic database querying Embase, PubMed, MEDLINE, Cochrane Library and PsycINFO. We searched original articles describing development, validation and measurement properties for screening tools on anxiety, QoL and food-related behaviors in IBD from 1975 to 2018. The primary outcome of interest was to evaluate the current measurements of the validated tools to identify whether a screening tool highlighting all above parameters exists for patients with IBD.

Results
Initial database search resulted in 5548 articles. After screening titles and abstracts, 168 were included. After full text review and deduplication, 23 validated scales were identified for use in IBD patients with respect to measuring anxiety, health-related QoL and food-related behaviors. There was substantial heterogeneity in IBD populations using the assessment tools (adult vs. pediatric, CD vs. UC, inpatient vs. outpatient). The breakdown of studies identified: 2 studies (8.6%) evaluated QoL and anxiety, 2 studies (8.6%) evaluated QoL and food-related behaviors. The remainder of studies individually assessed QoL, anxiety and food-related behaviors (47.8%, 26% and 8.6% respectively). None of the tools performed satisfactory to establish all three measurements in individuals with IBD.

Conclusions
Recent evidence suggests the presence of dietary hypervigilance in individuals with chronic GI conditions potentially leading to food restrictive behaviors impacting QoL. Screening models evaluating multivariable relations of anxiety in food-related behaviors and QoL in IBD is lacking. Efforts should be made to develop and validate a multi-assessment screening tool to aid in early identification of this prevalence in IBD patients to facilitate improved management outcomes.

Funding Agencies
None
A248 WHAT IS THE MINIMUM TRAINING STANDARD FOR POINT OF CARE INTESTINAL ULTRASOUND? A SINGLE CENTER, PROSPECTIVE, OBSERVATIONAL STUDY TO DEFINE TRAINING STANDARDS
Authors: Hansen T , Baraty B , Lu C , Tanyingoh D , Maaser C , Novak K L 
Published: February 2020

Abstract

Background
Higher demands for colonoscopy and magnetic resonance imaging (MRI) for long term disease monitoring can be expected as the prevalence of IBD continues to rise in Canada. As resources are perpetually constrained, timely access to effective monitoring strategies important to direct care are increasingly compromised. Intestinal ultrasound provides a cost-effective solution to these challenges. Intestinal ultrasound (IUS) is a patient-centered, accurate modality used during clinic by non-radiologists to enhance clinical decision making. Minimum IUS training standards have yet to be established.

Aims
The aim of this study is to report a single operator IUS performance characteristics after completion of 4 weeks of training with 100 completed supervised scans.

Methods
A single center, prospective, observational study over 4 years utilizing a convenience sample of patients presenting to the inflammatory bowel disease (IBD) clinic at the University of Calgary. A single operator compared IUS to gold standard (either colonoscopy, or alternative cross-sectional imaging) with sensitivity, specificity, positive and negative predictive value calculated for each year. Joinpoint regression was performed to analyze the trend for sensitivity and specificity over the study period.

Results
A total of 235 IUS were performed on 235 individuals diagnosed with IBD between 2013 and 2016. There were individuals with 26 ulcerative colitis and 209 persons diagnosed with Crohn’s disease. There was a non-significant increase in sensitivity and specificity point estimates over the 4 year period (Table 1). The sensitivity, specificity, positive predictive value, and accuracy of IUS in 2016 were 100% (95% CI: 81%-100%), 92% (95% CI: 62%-100%), 94% (95% CI: 72%-99%) and 97% (95% CI: 8%-100%) respectively.

Conclusions
IUS provision by a gastroenterologist having completed 100 supervised scans during training within an expert center is sufficient, resulting in high accuracy. Although there is some improvement over time, the trend towards improvement over time is not significant. This study provides evidence to inform IUS training programs with a minimum training standard benchmark, imperative with expanding demand and development of new expert centers.

Funding Agencies
None
A252 FUNCTIONAL ANALYSIS OF GENETIC AETIOLOGIES IN A DOWNSTREAM OF TYROSINE KINASE (DOK) PROTEIN IN THE PATHOGENESIS OF PEDIATRIC INFLAMMATORY BOWEL DISEASE (IBD)
Authors: Batura V , Guo C , Warner N , Leung G , Ricciuto A , Muise A 
Published: February 2020

Abstract

Background
IBD is a form of chronic inflammatory disorder of the gastrointestinal tract that arises due to genetic, environmental, immunological and microbial factors. The precise pathological mechanisms remain elusive. It is thought that the onset of pediatric IBD can largely be attributed to genetics. Muise lab, at SickKids, regularly screens children at the SickKids IBD clinic and through an international consortium to find possible genetic links to the disease. We report a patient at SickKids with biallelic mutations in DOK4 who has severe Crohn’s Disease along with other inflammatory conditions. Downstream of kinase (DOK) proteins are a family of adaptor molecules that serve as scaffolding proteins important in regulating cell signaling, especially in T cells. DOK4 has been shown to have negative regulatory effects on T cell activation but is also expressed across various other tissues where its function is yet to be determined. We predict that these mutations are causing immune cell dysregulation, which may be contributing to the patients IBD.

Aims
Through this study, we aim to enhance our understanding of the pathobiological mechanism of novel mutations in DOK4.

Methods
We have established T cell lines, expressing wild type and mutated DOK4, which will be used to perform functional tests, such as localization analysis through immunofluorescence and cytokine profiling, to check for T cell function. We have patient derived organoids, which will be used to assess changes in gut morphology using imaging techniques. We will also generate mutant zebrafish model that will be used to determine the susceptibility to colitis related to this mutation, disease progression and gut peristalsis using live imaging technology.

Results
Preliminary data shows variation in expression of the protein within patient derived peripheral blood mononuclear cells (PBMCs) compared to a healthy donor.

Conclusions
With this study, we hope to identify new therapeutic targets for patients with DOK4 mutations.

Funding Agencies
CIHRThe Leona M. and Harry B. Helmsley Charitable Trust
A255 EMPHYSEMATOUS CYSTITIS: A MARKER OF SEVERE CROHN’S DISEASE?
Authors: Hasan S , Salh B 
Published: February 2020

Abstract

Background
Emphysematous cystitis (EC) is characterized by the presence of air within the bladder walls, a complication of infection by gas producing organisms within the urinary system. However, rare cases of EC in the setting of infectious gastroenteritis have been reported suggesting an alternate etiology. We report a case of EC in the setting of severe Crohn’s colitis with no clinical evidence of urinary tract infection.

Aims
This report outlines the rare case of a patient with active Crohn’s disease who presented with bloody diarrhea, anemia, hypoalbuminemia and initial radiological evidence of EC. We expand on the subsequent clinical investigations, diagnosis, treatment plan and hospital course of the patient.

Methods
Case report, review of literature.

Results
A 43-year old female presented with a 2-month history of bloody diarrhea consisting of 8–12 bowel movements a day, 10 kg of weight loss and peripheral edema. She also had multiple ulcerated lesions on her abdominal wall and in the perianal region. Initial CT scan was significant for pancolitis, anasarca and EC. The follow-up CT cystogram, flexible cystoscopy and pelvic MRI confirmed the diagnosis of EC and ruled out any fistulous tracts in the pelvis including enterovesical/colovesical fistula. The patient did not report any urinary symptoms and the urinalysis was within normal limits. Infectious workup including urine and blood cultures, stool ova and parasite, stool C. difficile PCR, TB IGRA, Hep A, Hep B, Hep C and HIV serologies were all negative. Despite the paucity of infectious findings, the EC was empirically treated with an intravenous third-generation cephalosporin. Colonoscopy was significant for multiple ulcerated and hyperemic areas with pseudopolyps all throughout the right, transverse and left colon. Biopsies confirmed Crohn’s colitis with no evidence of granulomata or dysplasia, and were also negative for CMV immunohistochemistry. The perianal and abdominal wall lesions were suspected to be pyoderma gangrenosum although biopsies were equivocal. The colitis was initially treated with intravenous steroids followed by biologic therapy with Infliximab. While there was some interval improvement of symptoms, the in-hospital course was further complicated by a rectal bleed requiring surgical obliteration of the rectal artery, then a colonic perforation requiring total colectomy.

Conclusions
This is a rare case of EC in a patient with severe Crohn’s colitis. There was no clinical evidence of urinary tract infection or fistulising disease. According to our review, this is the first reported incident of EC in a patient with inflammatory bowel disease without any prior intra-abdominal surgeries. While active Crohn’s disease alone is a critical illness, we conclude that concomitant EC is a poor prognostic factor.

Funding Agencies
None
A261 MICROBIOTA PROFILES OF PATIENTS WITH MENTAL DISODERS DIFFER FROM THOSE OF HEALTHY CONTROLS
Authors: Rabbia V , De Palma G , Lu J , Verdu E , Collins S M , Anglin R , Surette M , Bercik P 
Published: February 2020

Abstract

Background
Mental disorders are the leading cause of disabilities worldwide, with depression and anxiety among the most common ones, affecting up to 1/3 of the worldwide population at least once in their lifetime. In both preclinical models and clinical studies, gut microbiota has been associated with altered behavior and anxiety or depression, respectively.

Aims
To investigate 1) whether the microbial profiles of patients with generalized anxiety disorder (GAD) and major depression disorder (MDD) differ from those of healthy controls (HC), and 2) whether specific bacterial taxa associate with GAD or MDD.

Methods
118 patients with primary GAD (n=82, 83.3 % female) or MDD (n=36, 62.9 % female) and 99 matched HC (66.6 % female) were recruited through the Anxiety Treatment and Research Centre. Anxiety, depression and stress levels were assessed by DASS-21 questionnaire. Stool samples were collected anaerobically and analysed for 16S rRNA gene sequencing through Illumina technique. The data was divided in 4 groups: 1) mental health disorder (MHD) combining GAD and MDD, 2) GAD, 3) MDD, and 4) HC. The data was analyzed following the pipelines of dada2 and QIIME2. RandomForest plugin for QIIME2 was used to investigate predictive characteristics of MHD, GAD or MDD microbiota. SPSS software v.23 was used to perform Spearman correlations and logistic regressions between microbial taxa and clinical scores.

Results
The mean anxiety score was 16.2 (severe anxiety) for GAD patients and 9.8 (moderate anxiety) for MDD patients; the mean depression score was 19.2 (moderate depression) for MDD patients and 16.0 (moderate depression) for GAD patients, while healthy controls averaged only 1.5 (normal anxiety) and 1.7 (normal depression) for anxiety and depression, respectively. The microbiota profile of the MHD group was predictive of the patients’ disease state with an 83.3% accuracy. In particular, increased relative abundance of Bacteroides ovatus and Bacteroides spp. and decreased relative abundance of Dialister spp. (Veilonellaceae), Haemophilus parainfluenzae and Bifidobacterium adolescentis, were predictive of MHD. Neither the GAD or MDD group microbiota profiles alone were accurate in the prediction of the patients’ disease state. There was a positive correlation between the relative abundance of Bacteroides spp. and a negative correlation between the relative abundance of Clostridium sensu stricto spp. and Sutterella, and the clinical scores of combined MDH and HC groups.

Conclusions
Our data suggest that patients with mental health disorders have different microbiota profiles compared to healthy controls. We have identified specific bacterial signatures that will inform mechanistic studies in gnotobiotic mouse models to investigate further the role of microbiome in mental disorders.

Funding Agencies
NIH