bluuebunny/crossref_metadata_2025 huggingface dataset:
Receptor activity-modifying protein 1 regulates the differentiation of mouse skin fibroblasts by downregulating α-SMA expression via suppression of high mobility group AT-hook 1 to promote skin wound repair
Authors: Song Ru , Ma Jiaxu , Yin Siyuan , Wu Zhenjie , Liu Chunyan , Sun Rui , Cao Guoqi , Lu Yongpan , Liu Jian , Su Linqi , Wang Yibing
Published: January 2025

Background

Skin innervation is very important for normal wound healing, and receptor activity-modifying protein 1 (RAMP1) has been reported to modulate calcitonin gene-related peptide (CGRP) receptor function and thus be a potential treatment target. This study aimed to elucidate the intricate regulatory effect of RAMP1 on skin fibroblast function, thereby addressing the existing knowledge gap in this area.

Methods

Immunohistochemical staining and immunofluorescence (IF) staining were used to measure the dynamic changes in the expression of RAMP1 and α-smooth muscle actin (α-SMA) in skin wound tissue in mice. Mouse skin fibroblasts (MSFs) stably transfected with Tet-on-Flag-RAMP1 overexpression (OE) and Tet-on-Flag control (Ctrl) lentiviruses were constructed for in vitro experiments. High mobility group AT-hook 1 (HMGA1) plasmids and α-SMA plasmids were used to overexpress HMGA1 and α-SMA, respectively. An α-SMA siRNA was used to silence α-SMA. Quantitative real-time polymerase chain reaction (qPCR), western blot and IF staining analyses were used to determine the mRNA and protein levels in the cells in different groups. A scratch wound healing assay was used to evaluate the cell migration ability of different groups. Cleavage under targets and release using nuclease (CUT & RUN) assays and dual-luciferase reporter assays were used to predict and verify the interaction between HMGA1 and the α-SMA promoter.

Results

RAMP1 and α-SMA protein expression levels in the dermis changed dynamically and were negatively correlated during dorsal skin wound healing in mice. RAMP1 OE in vitro inhibited the differentiation and promoted the migration of MSFs by decreasing α-SMA expression via the suppression of HMGA1, which was shown for the first time to bind to the α-SMA promoter and increase α-SMA transcription. RAMP1 OE also modulated extracellular matrix (ECM) synthesis and remodeling by promoting collagen III and MMP9 expression and decreasing collagen I, MMP2, and tissue inhibitor of metalloproteinases 1 expression.

Conclusions

Our findings suggest that RAMP1 OE decreases differentiation and promotes migration in MSFs by downregulating α-SMA expression via the suppression of HMGA1 and modulates ECM synthesis and remodeling, revealing a novel mechanism regulating α-SMA transcription, providing new insights into the RAMP1-mediated regulation of fibroblast function, and identifying effective nerve-related targets for skin wound repair.
EEG during dynamic facial emotion processing reveals neural activity patterns associated with autistic traits in children
Authors: Hill Aron T , Ford Talitha C , Bailey Neil W , Lum Jarrad A G , Bigelow Felicity J , Oberman Lindsay M , Enticott Peter G
Published: January 2025

Altered brain connectivity and atypical neural oscillations have been observed in autism, yet their relationship with autistic traits in nonclinical populations remains underexplored. Here, we employ electroencephalography to examine functional connectivity, oscillatory power, and broadband aperiodic activity during a dynamic facial emotion processing task in 101 typically developing children aged 4 to 12 years. We investigate associations between these electrophysiological measures of brain dynamics and autistic traits as assessed by the Social Responsiveness Scale, 2nd Edition (SRS-2). Our results revealed that increased facial emotion processing–related connectivity across theta (4 to 7 Hz) and beta (13 to 30 Hz) frequencies correlated positively with higher SRS-2 scores, predominantly in right-lateralized (theta) and bilateral (beta) cortical networks. Additionally, a steeper 1/f-like aperiodic slope (spectral exponent) across fronto-central electrodes was associated with higher SRS-2 scores. Greater aperiodic-adjusted theta and alpha oscillatory power further correlated with both higher SRS-2 scores and steeper aperiodic slopes. These findings underscore important links between facial emotion processing-related brain dynamics and autistic traits in typically developing children. Future work could extend these findings to assess these electroencephalography-derived markers as potential mechanisms underlying behavioral difficulties in autism.
How Epistemic Community Shapes Global Governance of AI in Military Domain?
Authors: Qi Haotian , Kamnerdrat Wanmingkhwan
Published: January 2025

The rapid development and applications of artificial intelligence (AI) technology in the military realm pose significant technical, ethical, and geopolitical risks. This article examines how members of the epistemic community, particularly Chinese academic and policy players, influence and shape the governance of AI’s military applications. Using social network analysis and comparative case studies, this research explores China’s interactions with its Western and non-Western counterparts, comparing their engagement in knowledge production as well as the construction of cooperative mechanisms. Our findings reveal that while Chinese stakeholders are quickly developing within the epistemic community, they lag behind their international counterparts in terms of knowledge production and their patterns of interaction within the global network. These Chinese members could strengthen their contribution by using a wider range of platforms to engage stakeholders and collaborate with international peers to produce knowledge about military AI. Our research is based on an original dataset of international academic and policy collaboration in AI’s military applications since 2013. This dataset will support and expand future research in this area.
Spatial and trophic ecology of the estuarine Little Tern Sternula albifrons
Authors: Cerveira Lara R , Pereira Jorge M , Veríssimo Sara N , Paiva Vítor H , Ramos Jaime A
Published: January 2025

Little Terns Sternula albifrons are estuarine birds extensively using increasingly urbanized coastal areas. Given the increased exposure of these ecosystems to anthropogenic activities, it is crucial to study the ecology of this species and interactions with human stressors to establish management measures. We collected regurgitates (n = 182) and blood samples (n = 48) to study Little Terns’ diet, health condition and isotopic niche, and individually tracked breeding adults (n = 9, during 2021 and 2022) at 2 sandy beaches 21 km apart and located within Ria Formosa Natural Park, Algarve, Portugal. Although both areas are adjacent to foraging coastal marine areas and an estuarine lagoon system with saltpans, they differ in human disturbance. We also compared breeding success and adult health measures between these 2 colonies. The main prey for both populations were Sand Smelts Atherina spp. and Gobies Pomatoschistus spp., and no differences in δ13C and δ15N stable isotopic values between colonies were found. Tracking data revealed that individuals foraged in the surroundings of their breeding colonies. Entrance Channels and Ocean were the most frequently used habitats for foraging by individuals from the less disturbed colony. Adults from the more disturbed colony foraged mostly in Entrance Channels. Overall, all individuals tended to forage during daytime. There were no differences in health measures of individuals between colonies, though breeding success at the most disturbed colony was comparably lower. It is important to protect both breeding areas and adjacent foraging habitats for Little Terns, particularly given the expected increasing pressure from human activities on coastal areas worldwide.
OP25 Young Researcher Award: Lymphocytic colitis can be transcriptionally divided into channelopathic and inflammatory lymphocytic colitis
Authors: Bhardwaj A , Münch A , Montague J , Koch S , Rosenstiel P , Escudero-Hernández C
Published: January 2025

Background

The pathobiology of the moderate/non-destructive inflammatory bowel disease (IBD) lymphocytic colitis (LC) is poorly understood. We aimed to define an LC-specific mucosal transcriptome to gain insight into LC pathology, identify unique genomic signatures, and uncover potentially druggable disease pathways.

Methods

We performed bulk RNA-sequencing of LC and collagenous colitis (CC) colonic mucosa from patients with active disease, and healthy controls (n=4-10 per cohort). Differential gene expression was analyzed by gene-set enrichment and deconvolution analyses to identify pathologically relevant pathways and cells, respectively, altered in LC. Key findings were validated using reverse transcription quantitative PCR and/or immunohistochemistry. Finally, we compared our data to a previous cohort of ulcerative colitis and Crohn’s disease patients (n=4 per group) to distinguish non-destructive from classic IBD.

Results

LC can be subdivided into channelopathic LC, which is governed by organic acid and ion transport dysregulation; and inflammatory LC, which is driven by microbial immune responses. Inflammatory LC displays an innate and adaptive immunity that is limited compared to CC and classic IBD. Conversely, we noted a distinct induction of regulatory non-coding RNA species in inflammatory LC samples. Moreover, compared to CC, water channel and cell adhesion molecule gene expression decreased in channelopathic LC, whereas it was accentuated in inflammatory LC and associated with reduced intestinal epithelial cell proliferation.

Conclusion

We conclude that LC can be subdivided into channelopathic LC and inflammatory LC that could be pathomechanistically distinct subtypes despite their shared clinical presentation. Inflammatory LC exhibits a dampened immune response compared to CC and classic IBDs. Our results point to regulatory micro-RNAs as a potential disease-specific feature that may be amenable to therapeutic intervention.

References

1.Miehlke S, Guagnozzi D, Zabana Y, et al. European guidelines on microscopic colitis: United European Gastroenterology and European Microscopic Colitis Group statements and recommendations. United Eur Gastroenterol J. 2021;9(1):13-37. doi:10.1177/2050640620951905

2.Bhardwaj A, Münch A, Montague J, Koch S, Rosenstiel P, Escudero-Hernández C. Lymphocytic colitis can be transcriptionally divided into channelopathic and inflammatory lymphocytic colitis. United Eur Gastroenterol J. 2024;12(6):737-748. doi:10.1002/ueg2.12531
OP27 Unlocking Inflammatory Bowel Disease subtypes: a deep dive into transcriptomics and Machine Learning
Authors: Saini N , Acharjee A
Published: January 2025

Background

Identifying molecular subtypes of IBD is essential to address inconsistencies in gene expression-based classifications, clinical variability, and treatment responses in Crohn's Disease (CD) and Ulcerative Colitis (UC). Building on prior efforts using different methods and datasets, this study aimed to derive and validate IBD subtypes using transcriptomics data and unsupervised machine learning.

Methods

This study analysed RNA-sequenced data from inflamed and non-inflamed intestinal biopsies of 2,490 adult IBD patients. K-means clustering, guided by Within Cluster Sum of Squares (WCSS) to determine the optimal ‘K’, identified subtypes within the dataset. Distinct clusters for UC and CD were derived from gene expression, with gene set enrichment and network analysis characterizing their features. Statistical tests (Chi-squared and ANOVA) linked these clusters to clinical data for UC and CD.

Results

K-means clustering revealed three distinct clusters in UC and CD, whose significant association with IBD severity (UC: p = 0.000263; CD: p = 0.007006) and IBD region (p < 0.000001) was determined by Chi Squared test. ANOVA showed age significantly influenced UC clusters (p = 0.0000345) but not CD clusters (p = 0.285). In UC, Cluster 1 focused on RNA processing, DNA repair, and rapid cell turnover, with upregulation of EXOSC genes and other related genes. Cluster 2 highlighted autophagy, stress response, and signaling processes, with upregulated expression of ATG13, VPS37C, and DVL2. Cluster 3 emphasized cytoskeletal stability over metabolic activity, marked by the upregulation of SRF, SRC, and ABL1. Notably, all UC clusters demonstrated upregulation of COX1, TMSB10, and ACTB. In CD, Cluster 1 was defined by cytoskeletal dynamics and reduced protein synthesis, with upregulated expression of CFL1, F11R, and RAD23A. Cluster 2 exhibited increased protein synthesis and stress response pathways, associated with aggressive disease phenotypes, with upregulation of MTREX, SART3, and GTF3C3. Cluster 3 prioritised cytoskeletal organisation over metabolism, featuring upregulation of TESK1, ABL1, and DVL2, along with other genes. Across all CD clusters, COX1, CDH1, and SF3B1 were consistently up-regulated.

Conclusion

Despite certain limitations, this study categorizes UC and CD into three transcriptomics-based subtypes, identifying meaningful IBD-associated patterns, key subtyping genes, and insights into the disease's complex pathogenesis. These findings may advance new therapeutic strategies and personalized medicine for patients with distinct IBD subtypes.
OP37 Delineating molecular and microbial signatures across disease severity in Inflammatory Bowel Disease in adult treatment naive patients through single-cell transcriptomic and microbiome profiling.
Authors: Leipner M , Rimmer P , Paun A , Cheesbrough J , Danilin S , Klein A , Scepanovic P , Sharma N , Trenkle P , Regan-Komito D , Iqbal A , Iqbal T
Published: January 2025

Background

Inflammatory Bowel Disease (IBD), including Crohn's disease (CD) and Ulcerative Colitis (UC), is a chronic inflammatory condition with complex etiology involving genetic, immune, microbiome, and environmental factors. This study aims to elucidate the cellular and microbial landscape of adult treatment-naive IBD patients with varying disease severity to identify potential disease mechanisms and therapeutic targets, presenting one of the largest single-clinic, IBD single-cell datasets to date.

Methods

Tissue biopsies were collected from inflamed and non-inflamed regions of the colon in UC patients and colon + ileum in CD patients. Using 10x Genomics single-cell RNA sequencing (scRNA-seq), we analyzed 1.4 million cells from 68 treatment-naive IBD patients (38 CD, 30 UC) and 70 symptomatic healthy controls. Shotgun metagenomic sequencing was performed on 36 IBD patients with matched scRNA-seq data to explore host-microbe interactions, and a further 49 patients with paired clinical data. At presentation, standard clinical metadata, Patient Reported Outcomes, post-biopsy treatment, and 6/12-month treatment response were collected for stratification across disease severity and response.

Results

Our analysis revealed distinct cellular populations and gene expression profiles between IBD patients and healthy controls along the disease severity axis. Specific immune cell subsets were significantly expanded in IBD patients, with inflammatory monocytes particularly prominent in those with higher disease severity, exhibiting upregulated expression of pro-inflammatory cytokines and chemokines. Additionally, a shift from IgA to IgG antibody-producing cells was observed along the disease severity axis, indicating a dysregulated immune response associated with disease progression. Fecal microbial community typing based on pathway abundance revealed two clusters defined by low or high abundance of Bacteroidetes phylum taxa. Integration with clinical metadata and transcriptional profiles showed that Bacteroidetes-high UC patients displayed lower levels of inflammation and up-regulation of interferon signaling in B cells and myeloid cells.

Conclusion

This comprehensive profiling of adult treatment-naive IBD patients provides valuable insights into the cellular and microbial factors contributing to IBD prior to treatment perturbation. Our findings offer a deeper understanding of the underlying disease pathology of IBD patients at the time of presentation along the axis of disease severity, expanding the dynamic range of IBD patient profiling and informing future therapeutic strategies that may more quickly put patients on the right treatment and better address the causes of IBD in addition to the inflammatory response alone.
DOP004 Proactive fecal calprotectin testing in ulcerative colitis: Initial results of the PROMOTE UC study
Authors: Rosenfeld G , Narula N , Leung Y , Afif W , Williams A J , Huang V , Singh H , McCurdy J , Jairath V , Lam M , Loomes D , Zezos P , Stewart M , Ma C , Ropeleski M , Ancheta-Schmit M , Tavakoli P , Khanna R , Lewis J , Colombel J F , Bressler B ,   Canadian Inflammatory Bowel Disease Research Consortium (CIRC)
Published: January 2025

Background

Fecal calprotectin (FC) is a biomarker of mucosal inflammation widely used in the management of inflammatory bowel disease (IBD). Given the convenience of home-based FC testing, we hypothesized that intensive, proactive FC monitoring would prevent symptomatic ulcerative colitis (UC) flares by detecting elevated FC levels prior to symptom onset and allowing clinicians to optimize therapy.

Methods

This prospective, multicenter, randomized, observational clinical trial evaluated intensive home-based, FC testing (IBDoc, Bühlmann Laboratories) versus usual care in adults with UC in clinical remission, defined as a modified partial Mayo score (mpMayo) ≤2 with no rectal bleeding. Patients in the intervention arm used the home testing kit every 2 months for 18 months or until symptomatic flare. The decision to modify therapy based on an elevated FC result was left to the discretion of the attending physician. Patients in the control group were followed according to usual care for 18 months or until symptomatic flare. Clinical data were collected at baseline and every 6 months. The primary endpoint was the time to symptomatic flare, defined as an increase in mpMayo ≥2 points from baseline or a rectal bleeding score ≥1.

Results

A total of 584 patients were followed at 14 sites in Canada and Australia until symptomatic flare or end of study. Demographics were similar between the control (n=293) and intervention arms (n=291), including mean age (41.9 vs 41.9 years), sex (46.8% vs 49.1% male), disease duration (1.61 vs 1.44 years), and mean mpMayo (0.39 vs 0.40) (Table). At study end, the flare rate of 30.9% in the intervention arm with proactive monitoring was slightly lower than the flare rate of 32.8% in the control arm (median follow-up 549 and 562 days, respectively). Overall, HR for flare in the intervention vs control arm was 1.01 (95% CI: 0.76, 1.34; p=0.96) (Figure). Risk for flare was also similar when analyzed by disease extent (proctitis: 1.13 [0.60, 2.14, p=0.71]; left-sided disease: 0.99 [0.61, 1.58, p=0.95]; pancolitis: 0.96 [0.60, 1.53, p=0.86]) or biologic use (biologics: 1.29 [0.86, 1.95, p=0.22], non-biologics: 0.78 [0.52, 1.18, p=0.24]) at baseline.

Conclusion

In this large, prospective clinical trial, approximately one-third of patients with UC in clinical remission developed a symptomatic flare within 18 months. Longitudinal proactive FC monitoring was not associated with a reduction in clinical flares. Further study is needed to determine the utility of home-based FC testing in guiding the management of UC.
DOP049 Vedolizumab is more effective at inducing histo-endoscopic healing in early Crohn’s Disease compared to late CD, with comparable healing rates regardless of disease location: data from the LOVE-CD trial
Authors: Lenfant M , De Hertogh G , Baert F , Bossuyt P , Löwenberg M , Clasquin E , Hoentjen F , Molnár T , D'Haens G , Vermeire S
Published: January 2025

Background

Achieving deep remission, encompassing clinical, endoscopic, and biological remission, is a long-term goal in Crohn's disease [CD] according to the STRIDE-2 guidelines. The role of histological remission in CD remains unclear. We evaluated the efficacy of vedolizumab [VDZ] for inducing histo-endoscopic remission in early versus late CD in the prospective, open-label LOVE-CD trial conducted in Belgium, the Netherlands and Hungary.

Methods

In the LOVE-CD trial, patients with moderate-to-severe disease (Crohn’s Disease Activity Index 220-450 and presence of ulcers at baseline endoscopy) received intravenous vedolizumab over a 52-week period. Early CD was defined as a diagnosis <24 months (treatment-naive or history of corticosteroid and/or immunomodulator use), and late CD as a diagnosis >24 months with prior anti-TNF exposure. Ileocolonoscopies were performed at three timepoints (baseline, week 26, week 52) during which biopsies were systematically collected in the terminal ileum and colon. Both the endoscopies and biopsies were centrally scored by experienced readers for the simple endoscopic score for Crohn's disease [SES-CD], Robarts’ histopathology index [RHI], and Geboes score [GS]. Endoscopic remission, histo-endoscopic mucosal improvement [HEMI], and histo-endoscopic mucosal remission [HEMR] were defined as SES-CD <4, GS ≤3B.1, and GS <2B.1 respectively. The association between histologic and endoscopic scores was studied with Spearman’s correlation coefficient. Intention-to-treat analysis with non-responder imputation was used to handle missing data.

Results

Of the 260 patients included in LOVE-CD (Table), 440 biopsies (336 colon, 114 ileum) from 179 patients at week 26, and 435 biopsies (323 colon, 112 ileum) from 177 patients at week 52 were analysed. There was a strong correlation between the endoscopic and histological assessment (colon r 0.69, ileum r 0.64, total r 0.64, p<0.001). Patients with early CD were more likely to achieve histological remission than patients with late CD at week 52 (38.4 % vs. 14.9%, p=0.00004). Endoscopic and histological healing rates at week 26 (endoscopic remission 36.9%, HEMI 30.4%, HEMR 24.2%) and week 52 (endoscopic remission 35%, HEMI 30%, HEMR 22.7%) were comparable. When looking at disease location, histological remission rates in the colon and ileum did not significantly differ (35.2% vs. 27.8%, p=0.15).

Conclusion

Vedolizumab was more efficient at inducing histo-endoscopic remission in early CD as compared to late CD, with most of the histo-endoscopic healing occurring during the first 26 weeks. Histo-endoscopic healing rates under vedolizumab were comparable in colonic and ileal disease. Histological scores (RHI, GS) are also applicable for the evaluation of ileal and colonic CD.
DOP055 Upadacitinib for induction of remission in pediatric Crohn’s disease: An international multicenter study
Authors: Yerushalmy-Feler A , Spencer E A , Dolinger M T , Suskind D L , Mitrova K , Hradsky O , Conrad M A , Kelsen J R , Uhlig H H , Tzivinikos C , Henderson P , Wlazlo M , Hackl L , Shouval D S , Bramuzzo M , Urlep D , Olbjørn C , D'Arcangelo G , Pujol-Muncunill G , Yogev D , Kang B , Gasparetto M , Rungoe C , Kolho K L , Hojsak I , Norsa L , Rinawi F , Sansotta N , Magen Rimon R , Granot M , Scarallo L , Trindade E , Velasco Rodríguez-Belvís M , Turner D , Cohen S
Published: January 2025

Background

Data on upadacitinib therapy in children with Crohn’s disease (CD) are scarce. We aimed to evaluate the effectiveness and safety of upadacitinib as an induction therapy in pediatric CD.

Methods

A multicenter retrospective study of children treated with upadacitinib for the induction of remission of active CD from 30 centers worldwide affiliated with the IBD Interest and Porto group of the ESPGHAN between 2022 and 2024. Demographic, clinical, laboratory, and imaging data as well as adverse events (AEs) were recorded at week 8 post induction. The analysis of the primary outcome was based upon the intention-to-treat (ITT) principle.

Results

One-hundred children (median age 15.8 [interquartile range 14.3–17.2] years) were included. All children were previously treated with biologic therapies and 89 with ≥2 biologics. At the end of the 8-week induction period, clinical response, clinical remission and corticosteroid- and exclusive enteral nutrition-free clinical remission (CFR) were observed in 75%, 56% and 52% of the children, respectively. Normal C-reactive protein was achieved in 68% of the children and fecal calprotectin (FC) <150 mcg/g in 58%. Combined CFR and FC remission was observed in 13/31 (42%) children with available data at 8 weeks [13% of the ITT population]. AEs were recorded in 24 children, the most frequent was acne (n=12). Two AEs (severe acne and hypertriglyceridemia) led to discontinuation of therapy.

Conclusion

Upadacitinib is an effective induction therapy for refractory pediatric CD. Efficacy should be weighed against the potential risks of AEs.
DOP064 Evaluation of pharmacological interventions in human Precision-Cut Intestinal Slices: insights into tissue-resident immunity in Inflammatory Bowel Diseases
Authors: Grieger K , Beneke V , Neuhaus V , Linge H , Kulik U , Gundert B , Aselmann H , Braun A , Hesse C , Dehmel S , Sewald K
Published: January 2025

Background

Inflammatory Bowel Diseases (IBD), including Crohn's Disease (CD) and Ulcerative Colitis (UC), are marked by dysregulated immune responses and significant T cell infiltration, leading to chronic intestinal inflammation. The global burden of IBD is increasing, yet its pathogenesis remains only partly understood. Our study aims to gain deeper insights into the immunological responsiveness of diseased tissue using ex vivo primary human intestinal tissue slices (Precision Cut Intestinal Slices; PCIS). PCIS are ultrathin tissue slices of the intestine of human donors that remain viable in culture, preserve intestinal architecture, and maintain all relevant intestinal cells including tissue-resident immune cells. Here, we specifically investigated the effects of the JAK1 inhibitor filgotinib and the calcineurin inhibitor pimecrolimus on tissue-resident immune cell responses in PCIS ex vivo.

Methods

PCIS from ileum and colon resections of Crohn’s disease (CD) and non-CD patients were stimulated ex vivo with Concanavalin A, anti-CD3/CD28 antibodies and IL-1β. Induced responses were modulated with anti-inflammatory treatments such as pimecrolimus and filgotinib for 24h. We evaluated tissue viability (LDH-assay), cytokine release (multiplex ELISA), and PCIS morphology (H&E-staining and Immune fluorescence).

Results

CD and non-CD intestinal tissue slices maintained viability and intestinal morphology, including disease-specific features in ex vivo culture such as immune cell infiltrates and higher abundance of CD4+ and CD8+ T cells. Ex vivo treatment with various immune stimulations revealed distinct disease-specific cytokine release patterns, with a primary focus on Th1 and Th17 responses, but also including other Th cell, innate and epithelial responses. Filgotinib (1-100 μM) dose-dependently inhibited the secretion of general proinflammatory and T cell-specific cytokines (maximal decrease e.g. IL-8: 8-fold, IL-6: 50-fold, IL-17A: 27-fold). Pimecrolimus (25 µM) specifically decreased T cell-specific cytokines (maximal decrease e.g. IL-2: 13-fold, IL-17A: 22-fold, IL-17F: 27-fold or IFNγ: 7-fold) while the release of general proinflammatory mediators like IL-8 was less affected.

Conclusion

Our data show the anti-inflammatory effects of filgotinib and pimecrolimus in primary intestinal tissue slices ex vivo. These effects include modulation of various T cell-associated cytokines and other molecules involved in the calcineurin and JAK-STAT pathways. Notably, the different treatments showed distinct effects on cytokine release profiles. This study underscores the potential of PCIS to elucidate the mechanisms of JAK1 inhibitors, advancing IBD treatment strategies and providing a test platform for new drugs in a complex human tissue system.
DOP068 Vedolizumab counteracts Parkinson’s disease-related brain pathology driven by gut-primed IBD T cells
Authors: Gerasimova E , Nicolai H , Eyvaz T , Shelyganov P , Winner B , Zundler S , Prots I
Published: January 2025

Background

The gut-brain axis has been increasingly recognized as a critical pathway linking Inflammatory Bowel Diseases (IBD) with neurodegenerative diseases. Chronic inflammation resulting from IBD can extend beyond the gut, and epidemiological data have also linked to the increased prevalence of neurodegenerative conditions like Parkinson’s disease (PD). However, the exact mechanisms underlying this interconnection are largely unclear. We therefore investigated how gut-primed T cells from IBD patients may drive neurodegenerative processes seen in PD.

Methods

Peripheral blood T cells from IBD patients were analyzed for co-expression of gut-homing integrin α4β7 and brain-homing integrin β1. Using gut-specific photoconversion in a Kaede T cell Transfer Colitis mouse model, we assessed the infiltration of gut-primed T cells into the brain tissue. To model and elucidate the interplay of T cells with the brain tissue ex vivo, we developed an innovative three-dimensional co-culture system involving human stem cell-derived Brain Organoids (hBrO) and peripheral blood T cells from IBD patients and controls. Vedolizumab, an anti-α4β7 integrin antibody, was used to assess its potential in blocking T cell accumulation and subsequent neurotoxicity in hBrO cultures.

Results

Gut-primed T cells from IBD patients co-expressed brain-homing integrins, suggesting that they are able to enter brain tissue. Clear evidence of gut-to-brain T cell trafficking was obtained in the Kaede T cell Transfer Colitis mouse model, confirming the capacity of gut-primed T cells to migrate across the blood-brain barrier under conditions of chronic intestinal inflammation. In the hBrO co-culture model, neurons expressed MAdCAM-1, and MAdCAM-1 expression in neurons was also found in single cell RNA sequencing of postmortem brain tissue. Consistently, α4β7+ IBD T cells accumulated specifically in the vicinity of neurons, leading to neuronal cell death and aSyn aggregation (a hallmark of PD) in hBrO. In line, Vedolizumab treatment reduced IBD T cell accumulation in hBrO and preserved neural cell integrity.

Conclusion

Our data, for the first time, show a T cell trafficking-driven axis of gut-to-brain communication with pathogenic potential for the central nervous system in the context of IBD. Moreover, our findings suggest that neurotoxic T cell-neuron interaction is mediated by MADCAM1 and α4β7 integrin. Thus, our study reveals a novel pathway, by which IBD T cells may contribute to neurodegeneration, linking chronic gut inflammation to brain tissue pathology and synucleinopathy. Blocking α4β7-mediated T cell-induced neurotoxicity and aSyn pathology presents a potential therapeutic approach for IBD patients at risk of neurodegenerative diseases such as PD.
DOP073 Gli1+ Mesenchymal Cells Drive Intestinal Fibrostenosis by Secreting SMOC2: A Biomarker and Therapeutic Target
Authors: Wang D , Zhao X , Zhang Y , Zou D
Published: January 2025

Background

Intestinal fibrostenosis in Crohn’s disease (CD) represents a significant clinical challenge, driven by the absence of reliable early biomarkers, limited therapeutic options, and high postoperative recurrence rates, all of which contribute to poor patient outcomes. This study aims to elucidate the role of Gli1+ mesenchymal cells in the pathogenesis of intestinal fibrostenosis and to explore their potential as a novel therapeutic target.

Methods

CD patients who underwent surgery for intestinal obstruction due to fibrosis were included in this study, and intestinal tissues were collected from both fibrotic and normal sites. Dextrose sodium sulfate (DSS)-induced and 2,4,6-Trinitrobenzenesulfonic acid (TNBS)-induced chronic colitis mouse models and their normal controls were also included. Single-cell RNA sequencing was performed on both human and murine tissues. Gli1-CreERT2; R26-tdTomato mice were generated for lineage tracing. Cell sorting combined with Smart-seq was used to isolate Gli1+ mesenchymal cells, revealing their pathogenic determinants.

Results

Single-cell RNA sequencing data and immunofluorescence staining from surgical patients revealed an enrichment of Gli1+ mesenchymal cells at fibrotic sites. Lineage tracing of Gli1+ mesenchymal cells demonstrated their proliferation and acquisition of a myofibroblast phenotype following chronic colitis. In vivo ablation of Gli1+ mesenchymal cells attenuated the severity of intestinal fibrosis. Compared to Gli1- cells, Gli1+ mesenchymal cells expressed a distinct gene expression profile, including markers of extracellular matrix (ECM)-related genes and Smoc2 (SPARC Related Modular Calcium Binding 2, encodes an extracellular matrix protein). Elevated Smoc2 expression was confirmed in both human and murine tissues, and Smoc2 knockdown further supported its role in promoting intestinal fibrosis. Mechanistically, Gli1+ mesenchymal cells establish a pro-fibrotic niche through the endocrine and exocrine functions of SMOC2, promoting ECM deposition and the activation of other fibroblasts. Additionally, a predictive and diagnostic model was established by assessing SMOC2 levels in tissue and plasma samples from CD patients with intestinal fibrosis, highlighting its potential as a biomarker for postoperative recurrence and as a therapeutic target.

Conclusion

Our results demonstrate that Gli1+ mesenchymal cells are key drivers of intestinal fibrostenosis, forming a pro-fibrotic niche through their unique secretion of the SMOC2 protein at fibrotic sites. SMOC2 could therefore be used as a predictive biomarker for postoperative recurrence and as a potential therapeutic target.
DOP095 IL-23-activated cells as response biomarkers for risankizumab in Crohn’s disease patients
Authors: Cao S , Ma K , Deepak P
Published: January 2025

Background

The interleukin (IL)-23 signaling is a vital pathway in Crohn’s disease (CD). IL-23 activates T-helper (Th) 17 cells, IL-17-secreting CD8 T (Tc)17 cells, γδ T cells, natural killer (NK) T cells, and group 3 innate lymphoid cells (ILC3) to secrete inflammatory cytokines including IL-17, IFN-γ, and TNF-α. Risankizumab is the first selective IL-23 antagonist approved for moderate-to-severe CD; however, no biomarker currently exists to predict response to this medication. Here, we characterized the phenotypes of IL-23-activated cells that predict response to risankizumab in CD patients.

Methods

Adult patients with active ileal/colonic CD were included (n = 28). Blood samples were collected before and after initiation of risankizumab. Peripheral blood mononuclear cells (PBMC) were isolated and cryopreserved for batch analysis. After stimulation with IL-23, PBMCs were stained using a pre-optimized and validated antibody panel for 37 lineage markers and cytokines for CyTOF. Cell clusters were resolved using UMAP (Figure 1), opt-SNE and FlowJo. Patients’ responses were evaluated 12-40 weeks after initiation of risankizumab based on clinical symptoms (e.g., CDAI) and endoscopic/radiographic assessment.

Results

Among the IL-23-activated cells, increased frequency of NKT cells was identified in the peripheral blood of non-responders before initiation of risankizumab (not shown). In pre-risankizumab PBMCs, we identified significantly elevated TNF-α in Th17 cells, Tc17 cells, NKT cells, and MAIT cells in non-responders vs. responders (Figure 2A). Similarly, in PBMCs 4 weeks after 1st risankizumab infusion, Tc17 cells, γδ T cells, NKT cells, and MAIT cells in non-responders produced significantly more TNF-α compared to those cells from responders. In addition, we observed increased IL-17A in multiple IL-23-activated cells in non-responders 4 weeks after starting risankizumab (Figure 2B). Peripheral and mucosal IL-23-activated cells from later non-responders before and 4 weeks after treatment spontaneously secreted TNF-α and IL-17A without stimulation. A similar correlation between therapeutic response and expression of TNF-α or IL-17A was not observed in ustekinumab, an anti-IL-12/IL-23 antibody (not shown).

Conclusion

CyTOF of patients’ PBMCs before and after starting risankizumab revealed previously unknown molecular signatures, especially augmented TNF-a production in multiple IL-23-activated cells, which predict non-response to risankizumab in CD patients. Those findings identified a novel mechanism of anti-IL-23 resistance, may help identify response biomarkers for IL-23 antagonists and provide a rationale for anti-IL-23/anti-TNF combination therapy in a subset of patients with refractory CD.
DOP102 Comparative effectiveness of risankizumab vs. ustekinumab in bio-naïve Crohn's disease patients: A real-world multicentre retrospective cohort study
Authors: Aldiabat M , Yarur A , Peyrin-Biroulet L , Loftus- Jr. E V , Colombel J F , Deepak P
Published: January 2025

Background

The SEQUENCE trial, in bio-exposed Crohn's disease (CD) patients, demonstrated non-inferiority for clinical remission at 24 weeks and superiority for endoscopic remission at week 48 for risankizumab (RZB) compared to ustekinumab (UST). However, this comparison has not been adequately explored in bio-naive CD patients. Our study aimed to compare the clinical effectiveness among bio-naive CD patients treated with RZB vs. UST in a real-world setting, using data from a US-based multicenter cohort.

Methods

This retrospective cohort study utilized data from the TriNetX US Collaborative Network, comprising sixty-eight tertiary institutions, to examine adult (≥18 years) bio-naive CD patients initiated on RZB or UST from June 17th, 2022 (FDA approval date) to October 31st, 2024. Patients with prior advanced therapy usage and those with approved rheumatologic disease indications for RZB/UST were excluded. A 1:1 propensity score matching (PSM) analysis was conducted to balance cohorts on demographics, baseline comorbidities, and indicators of CD severity, including CD phenotype, extraintestinal manifestations, concurrent corticosteroid use, hemoglobin, and body weight. Cox proportional hazards models estimated hazard ratios (HRs) in matched cohorts for one-year post-treatment composite outcomes (need for corticosteroids, hospitalizations, emergency room visits, surgeries) with a priori significance threshold of p < 0.05.

Results

The study sample included 1,630 bio-naive CD patients treated with RZB and 3,105 with UST. Among patients on UST, 658 (21%) were dosed every four to six weeks, after the first dose. Baseline characteristics of the PSM RZB and UST cohorts are shown in Table 1 with 1,568 patients in each cohort. After PSM (Figure 1), there were no significant differences in composite outcome (HR 1.077, p = 0.170), corticosteroid use (HR 1.099, p = 0.110), CD-related surgery (HR 1.231, p = 0.284), hospitalization (HR 0.947, p = 0.615) or ER visits (HR 1.041, p = 0.687). Additionally, no significant differences were observed in opioid use (HR 1.062, p = 0.366), switch to different advanced therapy (HR 1.097, p = 0.468) or elevated disease biomarkers, including CRP (>10mg/l, HR 1.186 p = 0.101) or fecal calprotectin (>150 µg/g; HR 1.109, p = 0.407) at 1-year follow-up.

Conclusion

In this real-world experience allowing for drug optimization, no difference was observed in bio naïve patients between UST and RZB as far as clinical remission and progression of CD.
P0005 Transcriptomic analysis reveals molecular differences between intestinal tuberculosis and ileal Crohn's Disease
Authors: Tang Q , Tian C , Ye L , Huang L , Cao Q
Published: January 2025

Background

Intestinal tuberculosis (ITB) and ileal Crohn's disease (iCD) are common chronic granulomatous diseases of the intestine, exhibiting similarities in clinical and histopathological features, which makes differential diagnosis challenging. We aim to explore the transcriptomic signature of ITB and iCD and identify potential biomarkers for differential diagnosis.

Methods

We prospectively enrolled seven iCD patients, seven ITB patients, and nine healthy donors. All participants underwent laboratory tests and colonoscopies. Biopsy samples from the ulcer margins of the ileocecal region were collected for bulk transcriptome analysis. Differentially expressed genes (DEGs; P-value<0.05, fold change>2) were selected for further analysis.

Results

Patients with ITB and iCD showed no statistical differences in routine blood tests, inflammatory markers, or nutritional indicators. Transcriptome analysis revealed 2,690 and 1,849 differentially expressed genes (DEGs) in ITB and iCD, respectively, compared to healthy controls. KEGG pathway enrichment analysis of DEGs that changed commonly or specifically in two diseases revealed shared activation of the IL-17 signaling pathway and upregulation of cytokine-cytokine receptor interactions in the iCD and ITB groups. Distinct changes in the iCD group included upregulation of virus-related responses, neuroactive ligand-receptor interaction, efferocytosis, and downregulation of bile secretion and cholesterol metabolism. In contrast, the ITB group showed activation of chemokine signaling and complement-related pathways and downregulation of tyrosine and retinol metabolism pathways. Further comparison of inflammatory cytokines, chemokines, and extracellular matrix (ECM) related genes among the three groups revealed that ITB patients exhibited a more robust pro-inflammatory response and more pronounced ECM remodeling than iCD patients. To identify diagnostic markers for ITB and iCD, we analyzed the DEGs that changed specifically in iCD or ITB, selecting the top 20 genes with the most tremendous fold changes. We found that genes such as CCL17, PDZK1, PLA2G12B, PRRX1, and ZNF488 showed significant differences in expression between the iCD and ITB groups. Receiver Operating Characteristic (ROC) Curve analysis demonstrated strong predictive value for these five genes in distinguishing ITB from iCD patients, with AUC values of 0.8214, 0.8367, 0.8750, 0.8571, and 0.9643, respectively.

Conclusion

Our study advances the molecular understanding of ITB and iCD, emphasizing distinct transcriptomic responses in the mucosal tissues of these conditions. Notably, five genes—PRRX1, PDZK1, PLA2G12B, ZNF488, and CCL17—exhibited strong diagnostic potential for distinguishing ITB from iCD.
P0011 Differential Impact of Diet and FMT on Colonic and Ileal Inflammation in humanized IBD Models
Authors: Deleu DVM- PhD S , Vidmar K , Troisi S , Wargo H , Masi L , Williams J , Artone S , De Santis S , De Salvo C , Petito V , Lopetuso L , Cominelli F , Pizarro T T , Scaldaferri F
Published: January 2025

Background

Fecal microbiota transplantation (FMT) has been shown to be a promising approach to treat active ulcerative colitis (UC). However, its use as a treatment for Crohn's disease (CD) is in its early stages, and scientific evidence for its effectiveness is currently limited. Also, diet plays a crucial role in shaping the gut microbiome and might affect the efficacy of FMT. Therefore, we aimed to investigate the additive effects of dietary interventions on multidonor FMT efficacy in preclinical humanized IBD models.

Methods

Age/sex-matched inbred ileitis-prone SAMP and C57BL/6 mice were treated with broad-spectrum antibiotics followed by humanization with either CD or UC multidonor patient-derived microbiota, and subsequently randomized to either Western, Mediterranean or Control diet in combination with healthy donor multidonor FMT or sham (n= 8/experimental grp; Fig1A). After one week, mice were subjected to DSS to induce acute colitis (7d, 2-2.5%), followed by another 7d of recovery. Disease activity indices (DAI) were determined and evaluated by area under the curve (AUC). Feces were collected every other day for 16S sequencing. At sacrifice, colonic lengths were measured, and blood and different tissues collected for histology, RNA and protein analysis.

Results

Independent from diet, FMT was shown to improve disease in SAMP mice (Fig1; P=0.047), whereas independent from FMT, SAMP and C57Bl/6 mice fed a Mediterranean diet showed significantly lower disease compared to both control and Western diets (all P<0.001). Dietary regimen appeared to primarily affect DAI, as among each diet no significant effect of FMT could be observed (Table 1). Nevertheless, C57Bl/6 mice receiving FMT in addition to Mediterranean diet had significantly higher survival rates (P=0.0096) compared to all other experimental groups.

Moreover, ileitis was more severe in SAMP mice receiving FMT (Fig1; P=0.0119), while the opposite was observed for colonic inflammation, with a trend towards less severe colitis (P=0.0729). Further, a trend towards less severe ileitis was observed for SAMP fed a Mediterranean compared to a Western diet (P=0.080), which was not observed for colitis. Microbial analysis and RNA sequencing data might explain these findings (ongoing).

Conclusion

In conclusion, independently from each other, both FMT and Mediterranean diet lower disease activity in two humanized mouse models of IBD. Although a trend towards increased efficacy is observed when combining FMT and diet, the additive effect of FMT within each diet seems to be rather limited. Yet, FMT may decrease the severity of colitis, while surprisingly it may worsen ileitis in SAMP mice. Hence, further analyses are required to elucidate the specific mechanisms behind these findings.
P0029 Abnormal glycerophospholipid metabolism in CCL2+DPP4+ mesenchymal stem cells may contributes to creeping fat fibrosis in Crohn's Disease
Authors: Yu A , Wu F
Published: January 2025

Background

Creeping fat (CF), an aberrant mesenteric adipose tissue (MAT), is characterized by abnormal lipid metabolism, inflammatory cell infiltration, and extracellular matrix (ECM) deposition in the intestines affected by Crohn's disease (CD). Our previous single-cell RNA-seq analysis identified a CCL2+DPP4+ subset of mesenchymal stem cells (MSCs) that accelerates the abnormal formation of CF. Building on these findings, this study investigates the role of CCL2+DPP4+ MSCs in CF fibrosis formation.

Methods

We employed H&E and Masson staining to assess histopathological changes and collagen deposition. Ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was conducted on CCL2+DPP4+ MSCs isolated from CF biopsies of CD patients (n=5) and MAT from controls without CD (n=5).

Results

We observed ECM deposition in CF and an upregulation of fibrosis-related genes (COL1A1, COL1A2, and FN1)（Figure1）. Analysis of our previous single-cell data revealed that CCL2+DPP4+ MSCs play a significant role in the abnormal accumulation of the ECM (Figure2).

Lipidomic profiling revealed differential lipid levels between the CF and healthy MAT（H-MAT） groups, with downregulation of specific lipids in CCL2+DPP4+ MSCs derived from CF, including PE (20:5_16:1), PC (14:0_22:6), and PE (16:1_14:0). KEGG pathway enrichment analysis highlighted significant enrichment in Glycerophospholipid metabolism. Phospholipids (PLs), crucial for membrane structure and function, were found to be regulated by key enzymes PISD(Phosphatidylserine decarboxylase) and LPCAT3(Lysophosphatidylcholine acyltransferase 3), which convert PS（phosphatidylserine） to PE（phosphatidylethanolamine） and integrate polyunsaturated fatty acids into lysophospholipids, respectively. Quantitative real-time PCR confirmed reduced expression of these enzymes and increased fibrosis gene expression in CF-derived CCL2+DPP4+ MSCs(Figure3).

Conclusion

Our findings suggest that the CCL2+DPP4+ MSC subset in CF exhibits abnormal glycerophospholipid metabolism, which is associated with decreased PISD and LPCAT3 expression. This may contribute to enhanced ECM formation and promotion of CF fibrosis. Understanding these mechanisms could lead to novel therapeutic approaches for CD.