bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

Abstract

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

Abstract

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

Abstract

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

Abstract

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

Abstract

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 &lt; 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 (&gt;10mg/l, HR 1.186 p = 0.101) or fecal calprotectin (&gt;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

Abstract

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&lt;0.05, fold change&gt;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

Abstract

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&lt;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

Abstract

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.