bluuebunny/crossref_metadata_2025_split huggingface dataset:
P0181 Modulating macrophage activity via NAAA pharmacological inhibition: a therapeutic strategy for intestinal fibrosis
Authors: Nani M , De Cicco P , Cattaneo F , Petrosino S , Tropeano F P , Miraglia M , Pagano E , Rinaldi M M , Izzo A A , Romano B ,   Francesca Paola Tropeano
Published: January 2025

Abstract

Background
Intestinal fibrosis, a frequent complication of inflammatory bowel disease, is characterized by stricture formation with no pharmacological treatment to date. N-acylethanolamine acid amidase (NAAA) is responsible for the hydrolysis of acylethanolamides (AEs, eg, palmitoylethanolamide and oleoylethanolamide). Here, we investigated NAAA and AE signalling in gut fibrosis.

Methods
NAAA and AE signalling were evaluated in human intestinal specimens from patients with stenotic Crohn’s disease (CD). Gut fibrosis was induced by 2,4,6-trinitrobenzenesulfonic acid, monitored by colonoscopy, and assessed by qRT-PCR, histological analyses, and confocal microscopy. Immune cells in mesenteric lymph nodes were analysed by FACS. Colonic fibroblasts were cultured in conditioned media derived from polarized or non-polarized bone marrow-derived macrophages (BMDMs). IL-23 signalling was evaluated by qRT-PCR, ELISA, FACS, and western blot in BMDMs and in lamina propria CX3CR1+ cells.

Results
In ileocolonic human CD strictures, increased transcript expression of NAAA was observed with a decrease in its substrates oleoylethanolamide and palmitoylethanolamide. NAAA inhibition reduced intestinal fibrosis in vivo. More in-depth studies revealed modulation of the immune response related to IL-23 following NAAA inhibition. The antifibrotic actions of NAAA inhibition are mediated by Mφ and M2 macrophages that indirectly affect fibroblast collagenogenesis. NAAA inhibitor AM9053 normalized IL-23 signalling in BMDMs and in lamina propria CX3CR1+ cells.

Conclusion
Our findings provide new insights into the pathophysiological mechanism of intestinal fibrosis and identify NAAA as a promising target for the development of therapeutic treatments to alleviate CD-related fibrosis.
P0182 The systemic administration of the anti-MCL1, S63845, reduces inflammation and fibrosis in a murine model of DSS-Chronic Colitis
Authors: Jarén A , Macías-Ceja D C , Burgos R , Cosín-Roger J , Cejudo-Garcés A , Patel J , Ortiz-Masiá D , Calatayud S , Barrachina M D 
Published: January 2025

Abstract

Background
A recent RNA sequencing analyses performed in ileal resections from complicated Crohn´s disease patients revealed the up-regulation of the anti-apoptotic molecule, MCL1 in fibrotic lesions. Immunohistochemical studies localized MCL1 expression in several cell types, including fibroblasts and leukocytes1. We analyse here the role played by MCL1 on inflammation and fibrosis in a murine model of chronic colitis.

Methods
C57Bl/6 mice (20–25 g) were administered intraperitoneally with Vehicle or S63845 (25 mg/kg), five days a week during the two DSS cycles (a cycle consists in 7 days drinking water with 2% DSS, followed by ten days drinking water). Body weight and Disease Activity Index (DAI) were monitored daily, and mice were sacrificed 34 days after the beginning of the experiment. A sample from the colon was paraffin embedded for both histological analysis (epithelial damage was quantified by the Obermeier score in H&E-stained slides and collagen deposition was quantified by the QuPath software in Syrius Red- stained slides) and immunohistochemistry (CD68, CD3, MPO, TUNEL and Cleaved-caspase 3). Other sample was used for RT-PCR studies. Unless otherwise mentioned, data are expressed as mean±SEM.

Results
DSS-Vh treated mice exhibited a reduction in body weight (16.9±1.0) and increase in DAI score (2.5±0.6) which reached a statistical significance (P&lt;0.001&, P&lt;0.01&, respectively) at day 11, compared with body weight and DAI at day 0 (19.4±0.4 and 0, respectively). In S63845-DSS treated mice, no significant changes in body weight and DAI were detected all along the experimental period when compared with the respective day 0. The administration of S63845, compared with Vh in DSS-treated mice induced: a) a reduction in epithelial damage (1.3±0.8 vs 1.7±0.4) and a significant diminution in the thickness of the submucosa (19.6±1.7 vs 42.4±3.2, respectively, P =0.0008*) and collagen deposition in the same area (median an IQ range: 54.2 (55.9-68.5) vs 61.9 (55.9-68.5) respectively, P = 0.051#). b) a lower immunostaining for both MPO and CD3; c) a similar immunostaining for CD68, TUNEL and Cleaved-caspase 3 which was very scarce for the last two and d) a lower mRNA expression of inflammatory cytokines (il1β, il6 and chi3l1), chemokines (cxcl1 and cxcl5) and profibrotic markers (col1a1, col3a1, col4a1, and tgfβ1). *t-test; #Mann-Whitnney; &Repeated measures ANOVA

Conclusion
The systemic administration of the MCL1 inhibitor, S63845 prevented the infiltration of both neutrophils and CD3 lymphocytes and failed to induce apoptosis in the affected intestine while it reduced fibrosis in a murine model of DSS-chronic colitis

References
1. Seco-Cervera M et al Biochim Biophys Acta Mol Basis Dis. 2024 Feb;1870(2):166966BBA
P0188 The Role and Mechanism of Ephrin B2 in Crohn’s Disease-Associated Intestinal Fibrosis
Authors: Hu S , Yu Q , Wang X , Xu D , Chen M , Chen Y 
Published: January 2025

Abstract

Background
Crohn’s disease (CD) is a chronic inflammatory bowel condition characterized by transmural inflammation that can lead to intestinal fibrosis and strictures. This study aimed to investigate the role of Ephrin B2, a member of the ephrin family, in the pathogenesis of CD-associated intestinal fibrosis.The primary objective of this study was to elucidate the expression pattern of Ephrin B2 in CD fibrosis and to explore its potential role in the modulation of autophagy and the AKT/mTOR signaling pathway, which were implicated in the fibrotic process.

Methods
The study used in vivo and in vitro methodologies to dissect the role of Ephrin B2 in intestinal fibrosis associated with Crohn’s disease. In vivo studies involved the generation of Ephrin B2 knockout mice and the induction of intestinal fibrosis using DSS-induced mice to assess the relationship between Ephrin B2 and intestinal fibrosis. In vitro studies utilized recombinant Ephrin B2 protein stimulation of CCD-18co cell lines to observe changes in autophagy-related proteins and the effects on fibroblast proliferation, activation, apoptosis, and collagen deposition. The study also investigated the impact of Ephrin B2 on signaling pathways such as PI3K/Akt/mTOR and RhoA/ROCK.

Results
Preliminary findings indicated that Ephrin B2 expression was significantly elevated in CD fibrosis patients and was associated with the modulation of intestinal autophagy genes. The in vivo model suggested a correlation between Ephrin B2 and the severity of intestinal fibrosis. In vitro studies showed that Ephrin B2 influenced the activation of fibroblasts and the deposition of collagen, potentially through the regulation of autophagy and the AKT/mTOR pathway.

Conclusion
This study provided evidence that Ephrin B2 might play a crucial role in CD-associated intestinal fibrosis. The findings suggested that targeting Ephrin B2 could be a novel therapeutic strategy for managing CD fibrosis. Further research is warranted to fully understand the mechanistic links between Ephrin B2, autophagy, and fibrosis in CD.
P0190 Hypoglycemia induces SLC7A11-dependent disulfidptosis to regulate intestinal inflammation in IBD
Authors: Chen C , Tao M , Fu Y 
Published: January 2025

Abstract

Background
IBD is an autoimmune disease related to intestinal mucosal immune dysregulation, and cellular metabolism is an important factor affecting intestinal immunity[1]. Previous studies have shown that the low-glucose environment inhibits the intestinal immune response to IBD, and the specific mechanism is unknown. Recently, a new mode of cell death has been discovered in the study of tumor cells, which is cytotoxic death caused by cysteine accumulation in cells with high expression of SLC7A11 in a low-glucose environment[2]. Therefore, we wanted to explore whether the phenomenon of disulfidptosis with low glucose environmental effects also exists in IBD and whether it has an impact on the regulation of intestinal inflammation in IBD.

Methods
We intend to explore the role and mechanism of SLC7A11-dependent disulfidptosis in low-glucose environment in regulating intestinal inflammation in IBD through mass spectrometry, multi-omics and cryo-electron microscopy, and further explore the molecular mechanism of regulating SLC7A11 at the protein structure level, so as to provide a solid theoretical basis for exploring new cell death modes in different glucose environments in IBD.

Results
We found that the expression of SLC7A11 in the colon of IBD patients and mice with DSS-induced colitis increased, the glucose content of the intestinal mucosa increased, and the genes related to disulfidptosis were down-regulated, suggesting that there may be inhibition of disulfide death in the high-glucose environment in IBD. We further showed that the low-glucose environment may induce disulfidptosis and inhibit inflammation in macrophages under inflammatory conditions, suggesting that the hypoglucose environment may inhibit IBD inflammation by promoting disulfidptosis.

Conclusion
The hypoglycemic environment in IBD may induce slc7a11-dependent disulfidptosis, thereby inhibiting intestinal inflammation.

References
1. Jung, J., et al. Metabolism as a guiding force for immunity. Nat. Cell Biol. 21, 85-93 (2019).
2. Liu, X., et al. Actin cytoskeleton vulnerability to disulfide stress mediates disulfidptosis. Nat. Cell Biol. 25, 404-414 (2023).
P0195 Immune reconstitution following autologous hematopoietic stem cell transplantation with cyclophosphamide-free mobilization for refractory Crohn’s disease
Authors: Saager E , Lutter L , Mahmmod N , Hoytema van Konijnenburg D , Delemarre E , van der Wal M , Oldenburg B , van Wijk F , Fidder H 
Published: January 2025

Abstract

Background
Autologous hematopoietic stem cell transplantation (aHSCT) has emerged as an effective therapeutic strategy for patients with severe chronic inflammatory conditions that are refractory to conventional treatment, including Crohn’s disease. The success of aHSCT is thought to be grounded in resetting the break of immunological tolerance, but the exact mechanisms underlying a favourable response remain incompletely understood.

Methods
We followed the immune reconstitution of 9 patients with severe refractory Crohn’s disease from one month until more than 2 years post-aHSCT. We performed flow cytometry to follow immune cell populations and functional T cells markers over time, in addition to Olink proteomics to study the inflammatory response in serum.

Results
Transplantation proceeded safely, without major adverse events in any of the 9 patients. aHSCT resulted in considerable improvement of disease status in 6 patients, with 3 patients classified as non-responders. After transplantation, we observed a strong change in the peripheral T-cell subset composition, with a reduction of the CD4/CD8 ratio and a reduction of naïve over effector memory (EM) T cells. Non-responders appeared distinct by a higher CD4/CD8 ratio and higher naïve over T-EM frequencies already before transplantation, and a smaller change in subset composition after transplantation. Remarkably, the local T-cell composition did not mirror the changes in the periphery, with an increased CD4/CD8 ratio in intestinal biopsies post-aHSCT, underscoring that a more detailed study of local versus peripheral responses to aHSCT is essential in aiding our understanding of its efficacy. Functionally, the changes in the peripheral T-cell profile were minimal and not clearly related to response. Although the pro-inflammatory profile was not altered, we did find a small but significant increase in the expression of regulatory markers.

Conclusion
aHSCT resulted in strong changes in the T-cell subset composition of Crohn’s disease patients, but only small changes in functional markers related to inflammation, regulation and homing. Non-responders could be distinguished at baseline by a different T-cell composition and also experienced less profound changes herein upon transplantation.
P0200 Evaluating the efficacy of tonsil-derived mesenchymal stem cells and intestinal stem cells-like cells in modulating fibrosis and inflammation in murine colitis models
Authors: Byeon J R , Song E M , Joo Y H , Choe A R , Park Y , Tae C H , Kim S E , Jung H K , Shim K N , Park Y S , Jung S A 
Published: January 2025

Abstract

Background
Current treatments in inflammatory bowel disease (IBD) primarily target inflammation, but effective anti-fibrotic treatments are limited. This study evaluates the therapeutic potential of tonsil derived mesenchymal stem cell (TMSCs) and TMSC induced intestinal stem cell-like cell (T-ISCs) in treating intestinal fibrosis and inflammation using two murine colitis models.

Methods
Two chronic colitis models were established using dinitrobenzene sulfonic acid (DNBS) and dextran sulfate sodium (DSS) in mice. TMSCs and T-ISCs were administered intraperitoneally, with mice divided into seven groups, including control and treatment groups. Colitis symptoms were assessed using disease activity index (DAI) and colon length. After euthanasia on day 30, colon tissues were collected for analysis. Fibrosis markers and inflammatory cytokines were assessed using real-time PCR, while fibrosis severity was evaluated through Masson’s Trichrome staining. In addition, immunofluorescence and western blotting was performed to confirm the fibrosis-related protein expression.

Results
The therapeutic effects of TMSCs and T-ISCs were confirmed in both DNBS and DSS colitis models. In the DNBS colitis model, both TMSCs and T-ISCs treatments significantly reduced DAI and restored colon length compared to the PBS group (colitis control group) (all p &lt; 0.05). TMSCs treatment markedly decreased three fibrosis markers, including COL1A1, Vimentin, and TGF-β, with statistical significance (p &lt; 0.01, p &lt; 0.01, and p &lt; 0.05, respectively), while T-ISCs significantly reduced only COL1A1 expression (p &lt; 0.05). Histological analyses using Masson’s Trichrome staining showed decreased fibrosis scores in both TMSCs- and T-ISCs-treated groups, and protein expression analysis via immunofluorescence and western blot also confirmed reductions in a-SMA, Vimentin, COL1A1, and TGF-β in both TMSCs- and T-ISCs-treated groups, with a particularly pronounced reduction observed in the TMSCs-treated group. In the DSS colitis model, both TMSCs and T-ISCs treatments significantly decreased the colon shortening compared to the PBS group (all p &lt; 0.05). Although fibrosis markers and scores showed no significant changes, inflammatory cytokine levels were markedly reduced in the DSS colitis model. Notably, T-ISCs demonstrated significant reductions in both IL-1β (p &lt; 0.01) and IL-6 (p &lt; 0.05).

Conclusion
TMSCs and T-ISCs demonstrate therapeutic effects in murine colitis models mimicking IBD, with fibrosis reduction in the DNBS model and inflammation reduction in the DSS model. These findings suggest that TMSC-based stem cell therapies are promising strategies for treating fibrosis and inflammation in IBD.
P0211 Characterization of drug-specific CD4+ T cells in patients with inflammatory bowel diseases
Authors: Weichberger A , Rios Martini G , Namuch Castro D M , Funcke N , Müller S , Marutz H , Sokolova M , Stamer N , Riffert J , Bergfeld A , Schulze Dieckhoff T , Nikolaus S , Aden K , Tran F , Schreiber S , Bacher P 
Published: January 2025

Abstract

Background
Biologicals are commonly used to treat chronic inflammatory diseases. Treatment failure is a major limitation for the efficacy of biologicals, which can be caused by the development of anti-drug antibodies. The formation of high-affinity anti-drug antibodies suggests the involvement of specific CD4+ T cell responses. However, the CD4+ T cell reaction against biologicals remains poorly characterized. Therefore, in this study we analyzed CD4+ T cells specific to various biologicals and investigated potential differences of drug-specific CD4+ T cells in patients with continued remission versus those with secondary nonresponses or allergic reactions.

Methods
Peripheral blood samples were collected at baseline and at week 6, week 14, week 22, week 30 after biological treatment. Drug-specific CD4+ T cells were analyzed using Antigen-Reactive T cell Enrichment (ARTE) directly ex vivo from human peripheral blood. To detect immunogenic regions, drug-specific CD4+ T cells were expanded and restimulated with individual peptides covering the whole variable chain of the therapeutic antibodies. Additionally, single-cell RNA sequencing was performed to investigate the gene expression profiles of drug-specific CD4+ T cells.

Results
Of the analyzed biological agents, the chimeric antibody infliximab showed significantly higher activation of reactive CD4+ T cells compared to other humanized or fully human biological agents. Notably, this response was characterized by a highly proliferative Th1 phenotype with evidence of long-term memory. In patients with secondary nonresponses or allergic reactions infliximab-specific CD4+ T cells showed elevated gene and protein expression of TNF-a, IFN-g and IL-21. Restimulation experiments indicated that drug-specific CD4+ T cell responses are driven by small immunodominant peptide regions.

Conclusion
Infliximab exhibits a higher activation of specific CD4+ T cell reactions compared to other biologicals, characterized by a proliferative Th1 response with long-term memory capabilities. Infliximab is a chimeric antibody consisting of murine variable regions, which likely contributes to its immunogenicity. The increased production of TNF-a, IFN-g and IL-21 in patients experiencing secondary nonresponses or allergic reactions highlights the potential role of drug-specific CD4+ T cells in treatment outcomes and formation of anti-drug antibodies. Overall, our data identifies differences in anti-drug CD4+ T cell reactions for different biologicals as well as different treatment outcomes which may contribute to enhance drug efficacy and improve long-term success with immunogenic biologicals.
P0220 Anti-TNFα, anti-integrin α4β7 and anti-IL23p19 biologic treatment dose optimization in a dextran sodium sulphate (DSS)-induced colitis mouse model
Authors: Suarez-Trujillo F , Orejudo M , Soleto I , Ramírez C , Baldán-Martín M , Martínez P J , Gordillo-Vélez C H , Cibrián D , Gisbert J P , Chaparro M ,   *These authors share senior authorship
Published: January 2025

Abstract

Background
Current immunotherapy for inflammatory bowel disease (IBD) treatment is based on anti-TNFα, anti-integrin α4β7, anti-IL12/IL23 and anti-IL23 antibodies that have demonstrated to be safe and effective. In vivo models are crucial for research with biologics, but the literature on doses of above-mentioned drugs in in vivo IBD models is scarce, highly variable and covers wide ranges. Thus, we aim to find the optimal dose for biologic treatment in a DSS-induced colitis model.

Methods
C57BL/6 mice were randomized into a control group and treatment groups receiving 3% DSS combined with varying doses of neutralizing antibodies against TNFα, integrin α4β7, and IL-23p19. Treated mice received 3% DSS ad libitum for 5 days and clean water for another 3 additional days (5 + 3 model). Neutralizing antibodies were injected intraperitoneally on days 2 and 5 of the experiment. Doses used for optimization were set for each dose at 100, 250 and 500 μg/mouse for anti-integrin α4β7 and anti-IL23p19 antibodies, and at 10, 25 and 50 μg/mouse for anti-TNFα antibody for covering ranges found in bibliography. Weight loss, fecal consistency and fecal occult blood parameters were measured every day of experiment to elaborate a disease activity index (DAI). Mice were sacrificed at day 8 and the colon was obtained for histological evaluation by H/E staining, and for extraction of lamina propria mononuclear cells (LPMCs). Longitude was also measured. Blood was obtained by intracardiac punction and peripheral blood mononuclear cells (PBMCs) were obtained. Distal and proximal colon sections were fixed in 4% paraformaldehyde and embedded in paraffin. 4 μm sections were hydrated and H/E stained for histological score assessment. Extracted LPMCs and PBMCs were stained for flow cytometry evaluation of inflammation markers.

Results
Proximal and distal colon of mice treated with two doses of 10 μg/mouse of anti-TNFα, 250 μg/mouse of anti-integrin α4β7 and 100 μg/mouse of anti-IL23p19 showed the greatest tissue conservation and repair (Figure 1). In addition, these groups had less weight loss throughout the experiment as well as a reduced DAI (Figure 2). LPMCs and PBMCs inflammatory markers were also decreased in these groups in comparison with the others. Slight differences were found in 250 and 500 μg/mouse anti-integrin α4β7 treated mice.

Conclusion
The optimal treatment doses of neutralizing anti-TNFα, anti-integrin α4β7 and anti-IL23p19 antibodies are set at two doses of 10, 250 and 100 μg/mouse respectively for this 5 + 3 model of DSS-induced colitis. By establishing precise dosimetry baselines, our work provides a robust framework for combinatorial therapies, which could unlock novel approaches for IBD treatment.