bluuebunny/crossref_metadata_2025_split huggingface dataset:
P045 MMP12 stimulates colonic afferents, and nociceptors through the activation of PAR1
Authors: Gupta R , Higham J , Raine T , Wright P , Bulmer D 
Published: January 2024

Abstract

Background
Abdominal pain is a cause of significant morbidity for people with colitis. We have previously shown that biopsy samples from people with Crohn’s disease (CD) are pro-nociceptive suggesting that peripheral nociceptor activation is a cause of pain in colitis. Furthermore, these effects were correlated with biopsy expression of macrophage metalloproteinase (MMP12), which was shown to stimulate mouse and human colonic nociceptors highlighting the potential utility of MMP12 inhibitors to act as visceral analgesics. Following on from this work we sought to determine the contribution of proteinase-activated receptor 1 (PAR1) to MMP12 mediated nociceptor activation.

Methods
MMP12 mediated sensory neuron activation: changes in the intracellular [Ca2+]i within thoracolumbar (T12-L5 spinal segments) dorsal root ganglia sensory neurons were examined in cells cultured overnight and loaded with Fluo-4-AM (30min) prior to imaging. Responses were measured to MMP12 or the PAR1 receptor agonists TRAP6 following pre-treatment with vehicle, the MMP12 inhibitor MMP408 or the PAR1 antagonist SCH79797. Afterwards cells were exposed to capsaicin (1 µM) followed by 50 mM KCl, to confirm the presence of nociceptors and neurons respectively by positive response.
MMP12 mediated colonic nociceptor activation: electrophysiological recordings were made of lumbar splanchnic nerve activity and of the isolated colorectum (ex-vivo) perfused with carbogenated Krebs solution, and responses determined following application with MMP12 alone or in the presence of respective MMP12 or PAR1 inhibitors.
All experiments were performed using tissue from male C57/B6 mice euthanised by rising concentration of CO2followed by exsanguination in accordance with Schedule 1 of the UK Animals Scientific Procedures act (1986).

Results
Application of MMP12 or TRAP6 produced a robust increase in [Ca2+]i levels in DRG sensory neurons classified as nociceptors by their co-sensitivity to capsaicin. These effects were significantly attenuated by pre-treatment with respective MMP12 or PAR1 inhibitors, with the response to MMP12 also being inhibited by PAR1 antagonist pre-treatment. Consistent with this pattern of nociceptor stimulation, the application of MMP12 also a robust activation of colonic afferents that was abolished by inhibition of MMP12 or PAR1 activity.

Conclusion
Findings demonstrate that MMP12 stimulates colonic afferents, and nociceptors through the activation of PAR1.
P074 Inflammatory Environment-Induced Transcriptomic Alterations in Crohn's Disease Adipose Stem Cells
Authors: Montfort-Ferré D , Boronat-Toscano A , Sánchez-Herrero J F , Caro A , Menacho M , Vañó-Segarra I , Martí M , Espina B , Pluvinet R , Cabrinety L , Abadia C , Ejarque M , Nuñez-Roa C , Maymo-Masip E , Sumoy L , Vendrell J , Fernández-Veledo S , Serena C 
Published: January 2024

Abstract

Background
Crohn’s disease (CD) is characterised by expansion of mesenteric adipose tissue, called creeping fat, which seems to be directly related to disease activity. Adipose stem cells (ASCs) isolated from the creeping fat of CD patients exhibit dysfunction, featuring impaired adipogenesis and an intensely pro-inflammatory phenotype. This study aims to explore the transcriptome of ASCs isolated from active and inactive CD subjects in comparison with healthy subjects, seeking key markers of this dysregulation.

Methods
Patients were recruited at Hospital Joan XXIII of Tarragona and Hospital Vall d’Hebron of Barcelona in accordance with the principles of the Helsinki Declaration. Transcriptome profiling was performed in ASCs isolated from adipose-tissue biopsies of visceral origin: CF and hMES in Crohn subjects (n=7, each) and hMES in inactive CD patients (n=7) and healthy control subjects (n=7). Groups were matched by age, gender, and BMI. Finally, we examined the pathways involving the differentially expressed gene (DEGs) in ASCs isolated from patients with CD with different clinical activity by gene set enrichment analysis (GSEA).

Results
Patients with CD across different clinical activity stages exhibited similar transcriptome patterns, indicating persistent ASC dysregulation even during remission state (Fig 1A). ASCs isolated from both creeping fat and mesenteric adipose tissue distant from intestinal damage displayed similar dysregulation, implying a global dysregulation of all mesenteric fat in CD. Specifically, transcriptomic analysis identified significant dysregulation of the NK2 Homeobox 3 (NKX2-3) gene in ASCs from active CD patients. Although higher NKX2-3 expression has been associated with B cells and intestinal tissues in CD1-3, our study is the first to link elevated expression of this gene also to creeping fat. Pathway enrichment analysis revealed significant enrichment in oxidative stress and immune response pathways in active CD (Fig 1B). Furthermore, our findings indicated also an elevated NKX2-3 expression in ASCs of inactive CD compared to controls, with oxidative stress pathway up-regulation (Fig 1C). Notably, no significant DEGs were found between ASCs from active and inactive CD groups (Fig 1D), suggesting a similar profile in both states.

Conclusion
Our study reveals profound ASC dysregulation in CD, in both active and inactive phases. Elevated NKX2-3 expression, previously associated with increased risk of complications in IBD, is now demonstrated in the mesenteric adipose tissue, persisting even in inactive CD patients.
Ref:
1. Yamazaki K, et al Gut. 2009 Feb;58(2):228-32.
2. Yu W, et al. J Crohn’s Colitis. 2009 Sep;3(3):189-95.
3. Hong SN et al. Gut. 2016 May;65(5):788-96.
P107 Comparing the Colonic Tissue Proteome of Active and Inactive Ulcerative Colitis
Authors: Penner R , de Almeida L , Rosentreter R , Hirota S , Dufour A , Lu C 
Published: January 2024

Abstract

Background
Ulcerative colitis (UC) is an inflammatory bowel disease that begins at the rectum and can advance proximally and continuously throughout the colon. In this pilot study, we aim to differentiate the protein expression between active and inactive patients, comparing the same tissue region of the colon pairwise between active and inactive UC. Additionally, we are looking to build on previous work to determine which proteins are differentially proteolyzed in active UC and postulate whether this alternative post-translational processing significantly correlates to disease activity. Here, we aim to use a multi -omics approach to spatially profile the entire large intestine.

Methods
We used an N-terminomics proteomic approach to determine whether proteins are differentially expressed and proteolytically processed in all colon segments of patients with active and inactive UC. Colon tissues were retrieved from patients and classified according to disease activity. Five active and five inactive UC patients were compared from each of the colonic regions (rectum, sigmoid, transverse, ascending colon). Samples were quantified by liquid chromatography and tandem mass spectrometry (LC-MS/MS). Peptides were identified and matched at a 5% false discovery rate (FDR). Boxplot analysis was used to determine the upper and lower threshold for protein fold changes between conditions, and various bioinformatic software (MetaScape, StringDB, TopFINDer) were used to analyze the data.

Results
Primary analysis discovered a combined 36 statistically changing proteins in patients with active disease, including several that have previously been published and others that are direct interactors with known biomarkers for active UC, including calmodulin and S100A9. Interestingly, S100A9 was translated significantly more in the transverse colon of patients with active UC but was not in other colonic regions. Protease activity was noticeably different in active and inactive UC, in addition to having unique activity dependent on the region of the colon from which the samples were acquired. For example, delta-catenin was processed more in active UC in the rectum and transverse colon, whereas it’s processing was enriched in inactive UC in the sigmoid colon.

Conclusion
While this pilot study is incomplete, the promising initial results exhibit the power of a proteomics and N-terminomics approach to elucidate the mechanisms that contribute to UC pathogenesis. There appears to be a pattern in how proteins and their altered proteolysis leads to an increased likelihood of active UC.
P109 Organic Cation Transporter (OCTN)-1 variants shape innate immunity and predict individual response to vedolizumab In ulcerative colitis patients
Authors: Capobianco I , Puca P , Masi L , Petito V , Pafundi P C , De Felice M , Fidaleo M , Di Vincenzo F , Del Chierico F , Delogu G , Palucci I , Laterza L , Pizzoferrato M , Schiavoni E , Napolitano D , Turchini L , Amatucci V , Strazzeri M , Pugliese D , Putignani L , Armuzzi A , Lopetuso L R , Gasbarrini A , Pani G , Scaldaferri F 
Published: January 2024

Abstract

Background
Limited evidence is available about predictors of response to specific medications for ulcerative colitis (UC). OCTN1, an organic cation transporter, with its variants, could modulate the inflammatory response and potentially modify drug response to immune-modulatory therapy (anti-TNFα and vedolizumab), perhaps predicting individual response.

Methods
We first confirmed the ability of OCTN1 to differentially modulate IL1β secretion under different stimuli in a human leukemia monocytic cell line (THP-1) and in human peripheral blood mononuclear cells. We therefore prospectively enrolled a cohort of UC patients starting either anti-TNFα or vedolizumab, determined their OCTN1 genotype and evaluated their clinical features at baseline and clinical response to therapy or disease remission (according to clinical Mayo score) after six weeks (T1) and six months (T2). We finally enrolled a cohort of UC patients in clinical and endoscopic remission (according to full Mayo score) under immune-modulatory therapy (anti-TNFα or vedolizumab) for at least two years.

Results
OCTN1-deficient THP1 cells showed reduced IL-1β secretion in response to peptidoglycan (PGN) and Fusobacterium nucleatum (Fn), whilst primary monocytes bearing OCTN1 503F variant displayed an increased production of IL-1β in response to PGN and live bacteria. We therefore enrolled 90 patients, 50 starting anti-TNFα and 40 vedolizumab. Patients with 503F genotype showed higher rates of pancolitis as compared to 503L genotype. Among patients with 503L genotype, vedolizumab results highly associated with long-term response (OR 8.92, 95%CI 0.50-11.0; p=0.019), whilst conversely co-presence of 503F genotype and vedolizumab therapy disclosed an opposite role on the long-term response rate (OR 0.02, 95%CI 0.00-0.81; p=0.038).The population of UC patients in clinical and endoscopic remission was composed by 92 patients, 63 under anti-TNFα and 29 under vedolizumab. Of note, similar trends of disease extension were observed across genotypes: higher rates of pancolitis were observed in the 503F population as compared to 503L (62.5% vs 44%). Furthermore, among patients in remission with 503F genotype, the percentage of subjects receiving vedolizumab was sensibly lower than among patients with 503L genotype (87.5% anti-TNFα vs12.5% vedolizumab among 503F genotype; 56% anti-TNFα vs 44% vedolizumab among 503L genotype).

Conclusion
Mutated OCTN1 genotype (503F) favors an increased IL1β secretion from human stimulated leukocytes, thus suggesting a worse disease course with higher rate of pancolitis and lower response to vedolizumab as compared to 503L.
P113 Differentiation between active and quiescent Ulcerative Colitis using a novel wearable patch with self-adhesive dry electrodes: discriminating the role of heart rate variability
Authors: Metselaar P , van den Broek T J , Kamstra R J M , Uzunbajakava N , D'Haens G R , van den Brink W , te Velde A A , Löwenberg M 
Published: January 2024

Abstract

Background
The autonomic nervous system (ANS) maintains homeostasis of the gastrointestinal tract through the brain-gut axis. Previous studies have shown a dysregulated balance between the sympathetic and parasympathetic nervous system in Ulcerative Colitis (UC) patients, in particular during and preceding active disease. This leads to decreased vagal nerve activity and manifests in reduced heart rate variability (HRV). HRV could, therefore, potentially serve as quantitative non-invasive biomarker to differentiate active from quiescent UC. We aimed to increase our understanding of autonomic function in UC, by investigating a non-invasive wearable patch device measuring HRV to assess the difference in parasympathetic and sympathetic activity between active and quiescent UC, while correcting for HRV-modulating factors depression, anxiety and fatigue.

Methods
A proprietary vital signs patch with self-adhesive dry electrodes was used to acquire ECG (single lead) and bio-impedance (2 leads) in 26 UC patients for 72 hours continuous home monitoring, to calculate heart rate, respiratory rate and HRV. Fifteen patients had quiescent UC (SCCAI ≤ 2, CRP &lt; 5mg/l, faecal calprotectin &lt; 250µg/g) and eleven had active disease (SSCAI &gt; 2, CRP &gt; 5mg/l, faecal calprotectin &gt; 250µg/g). Patients filled out the Multidimensional Fatigue Inventory and Hospital Anxiety and Depression Score questionnaires on fatigue, depression and anxiety, kept sleep diaries and provided feedback on patch usability. Time-domain (pNN50, RMSSD, for abbreviations see Figure 1), frequency-domain (high (HF), low (LF), LF/HF ratio), as well as non-linear (SD1, SD2, SD) HRV metrics were calculated for 5-minute segments during self-reported nocturnal sleep periods.

Results
No significant differences were found in fatigue, depression, and anxiety in the active and quiescent UC patient groups. Six HRV measures were significantly lower in active than in quiescent disease (p&lt;0.001, Figure 1), indicating lower HRV and lower vagal parasympathetic tone. LF/HF ratio did not differ (p=0.242, Figure 1), reflecting no difference in ANS balance. The patch caused skin irritation in 72% of the patients. Nevertheless, 69% of patients indicated they would wear it again.

Conclusion
A wearable vital signs patch, utilizing ECG, is able to distinguish active from quiescent UC in patients matched for fatigue, anxiety, and depression. HRV metrics across all three domains, except LF/HF ratio, were reduced in active UC, indicating reduced vagal nerve activity. Pending further longitudinal validation and clinical integration, it is envisioned that continuous HRV monitoring can support early and non-invasive detection of relapses in UC, allowing timely intervention and prevention of severe symptoms.
P115 Ustekinumab in DEVELOP: A safety analysis from an Inflammatory Bowel Disease multicenter, prospective, long-term registry of paediatric patients
Authors: Koletzko S , Veereman G , Hyams J , Dubinsky M , Godwin B , Busse C , Strauss R , Volger S , Wang Y , Griffiths A , Colletti R B , Kugathasan S , Markowitz J , Winter H S , Escher J , Baldassano R N , Ruemmele F , Faubion W , Gold B 
Published: January 2024

Abstract

Background
DEVELOP is a multi-center, global, prospective, observational, longitudinal registry of long-term safety of infliximab (and other treatments) in paediatric patients (pts) with inflammatory bowel disease (IBD) &lt;18 years (yrs) at diagnosis. Over the course of follow-up, many pts required changes in therapy. Ustekinumab (UST) is approved for treatment of Crohn’s disease (CD) in adults and is currently being evaluated for safety/efficacy in paediatric pts. This report focuses on assessment of safety of UST in paediatric pts with CD from DEVELOP.

Methods
Data included paediatric pts treated with UST from 31 May 2007 through 30 June 2022; data are collected every 6 months. The analyzed population included: all paediatric pts (all pts), pts weighing &lt;40kg (&lt;40kg group), and pts weighing ≥40kg (≥40kg group). Assessments included medical and treatment history, UST exposure, adverse events (AEs), serious AEs (SAEs), CD-related SAEs leading to hospitalization, IBD surgeries, serious and opportunistic infections, malignancies, and deaths.

Results
A total of 150 pts received UST (49.3% females); 31 in the &lt;40kg group and 119 in the ≥40kg group (Table 1). Most treated pts (92.0%) were 12-17yrs. The majority (91%) of pts were initially dosed with UST every 8 weeks (q8w); at the last known dose, 64% received UST q8w and 22% q4w. Mean (SD) age of initial UST exposure was 15yrs (2.19). Prior IBD surgery was reported in 65.3% pts. The majority of pts (98%) had prior biologic therapy and 72% received ≥2 biologics prior to UST. Overall, 37/150 (24.7%) of pts discontinued UST during follow-up, with a mean (SD) time to discontinuation of 16.7 (17.00) months; 84/150 (56.0%) were exposed to UST for ≥24 months.
Rates (events/100 pt-yrs) of AEs were higher in the ≥40kg group (177.62) than in the &lt;40kg group (101.27). Rates of SAEs were slightly lower among the ≥40kg group (25.42) vs &lt;40kg (32.67). There were no SAEs related to infusion or injection site reactions. Rates of serious infections and CD-related hospitalizations were low and similar between groups, and rates of IBD-related surgeries were similar between groups (Figure 1). One malignancy (malignant carcinoid tumor) was reported in a female (18yrs) in the ≥40kg group with a history of prior therapy with vedolizumab, methotrexate, and 6-MP. No deaths or opportunistic infections were reported.

Conclusion
The safety profile observed for this off-label use of UST in paediatric CD pts in DEVELOP was similar to that of the treated adult population, despite most paediatric pts having prior biologic exposure. No new safety signals are identified in this paediatric population.
P120 A novel population of classical monocytes is expanded in CD and defined by an upregulation of genes relating to mononuclear cell differentiation and response to microbes
Authors: Gadhok R , Hornsby E , Kok K B , Lindsay J O , Stagg A J 
Published: January 2024

Abstract

Background
In Crohn's disease (CD), differentiation of classical blood monocytes to intestinal macrophages in the mucosa generates inflammatory rather than regulatory cells. Microbial products and cytokines can systemically reprogramme monocytes via effects on myelopoiesis. We hypothesised that in CD transcriptional changes occur in monocytes prior to their recruitment to the intestine which predispose to pro-inflammatory signatures in the mucosa. Here, we used single cell RNAsequencing (scRNAseq) to explore monocyte heterogeneity and identify CD-associated changes in gene expression profiles.

Methods
PBMCs were isolated from newly diagnosed CD patients (n=10) and matched healthy controls (HC, n=10). scRNAseq was performed using the 10x Genomics Chromium 3’ assay at a target capture rate of 10,000 cells/sample and sequencing at ~50,000 reads/cell. A dataset was generated using Cell Ranger and following unsupervised clustering (using Seurat) the CD14+ and CD16+ monocyte populations were selected. Data were further analysed using Seurat, DeSEQ2, g:Profiler, GSEA and Cytoscape.

Results
In addition to non-classical and intermediate monocytes, five transcriptionally distinct clusters of classical monocytes were identified in both CD patients and controls. One cluster (‘cluster 3’) was significantly increased in abundance in CD; other clusters were present at similar frequency in health and CD. Cluster 3 was defined by gene ontology (GO) pathways related to response to bacteria, cell migration, lymphocyte proliferation and response to oxidative stress. In CD compared with health, the most upregulated genes included those encoding EGR (early growth response) proteins 1-3 that play a key role in the functional plasticity of monocytes, and the key inflammatory cytokine IL-1β, as well as cytokines and regulators of their signalling (e.g. CXCL2/3, SOCS3). Overall, in Cluster 3, GO pathways positively enriched in CD related to microbial response signalling, canonical WNT signalling and mononuclear cell differentiation.

Conclusion
Classical monocytes are heterogeneous and include a novel subset that is expanded in CD. In this population, the expression of genes and pathways related to functional plasticity, response to microbes and inflammatory activity are enhanced in CD. These data support a systemic reprogramming of monocytes in CD that predisposes to inflammatory activity upon recruitment to the intestine.
P122 Digital spatial profiling reveals the predictive characteristics in patients with immune checkpoint inhibitor-induced colitis
Authors: Shi Q , Zeng Y , Zhou Q , Liu X , Tang H , Lu B , Qian J , Chen M , Xu Y , Wang M , Tan B 
Published: January 2024

Abstract

Background
The increasing use of immune checkpoint inhibitors (ICIs) against cancers is inevitably accompanied with immune-related adverse events (irAEs), and irAE colitis is one of the common types. The overactivation of CD8+ T cells plays a central role in irAEs. However, the local mechanisms related to irAE colitis remain unclear. Hence, we aimed to explore the transcriptional characteristics related to CD8+ T cell responses of irAE colitis.

Methods
Colon biopsy samples were collected from 6 irAE colitis patients and 5 non-irAE controls after ICIs treatment. Leveraging digital spatial profiling, 23 regions of interest (ROI) enriched with CD8+ T cells were selected and divided into 3 groups, which were non-irAE, pre-treatment irAE and post-treatment irAE. Differentially expressed genes (DEGs), protein-protein interaction (PPI) and weighted gene co-expression network analysis (WGCNA) were performed to identify the hub genes, and they were further verified by external single-cell RNA-seq data. Also, the immune infiltration analysis was performed (Figure 1A).

Results
Principal component analysis (PCA) indicated similar distribution among the 3 groups (Figure 1B). The comparisons of non-irAE vs. pre-treatment irAE, and pre-treatment irAE vs. post-treatment irAE yielded 105 and 210 DEGs respectively. WGCNA resulted in 14 clustering modules, with MEturquoise and MEmagenta exhibiting the highest correlation scores (Figure 1C). Importantly, 174 key genes were identified and prominently associated with various mitochondrial processes e.g. oxidative phosphorylation (Figure 1D). PPI analysis revealed a sub-module consisting of 22 nodes in MCODE. Further overlap analysis of CytoHubba and DEGs pinpointed 9 hub genes. Remarkably, COX5B was found at the intersection of both groups (Figure 1E). External single-cell RNA-seq data verified that all the hub genes were significantly highly expressed in the irAE colitis group except COX6A1 (Figure 1F). Besides, immune infiltration analyses revealed the higher levels of endothelial cells (P = 0.043) and fibroblasts (P &lt;0.01) by SpatialDecon, and activated dendritic cells (P &lt;0.001) by xCell in pre-treatment irAE, compared to non-irAE (Figure 1G). Further, immune cells correlation analysis demonstrated highly similar patterns of endothelial cells, fibroblasts, pDCs, and macrophages (Figure 1H).

Conclusion
COX5B related to mitochondrial processes characterized irAE colitis. Expansion of dendritic cells and stromal compartments including endothelial cells and fibroblasts may contribute to irAE development.