bluuebunny/crossref_metadata_2025_split huggingface dataset:
P0151 Multi-modal single cell sequencing identifies CCR9 expression in lamina propria with highest protein expression on CD4 tissue resident memory cells in the inflamed ileum in Crohn’s disease
Authors: Hamilton R , Collin J , Keith B , Gehrmann U , Lindkvist B , Lundin S , Eberhardson M , Marks D , Platt A , Muthas D , Corridoni D , Bouma G 
Published: January 2025

Abstract

Background
Crohn’s Disease (CD) is a chronic condition that can impact any part of the gastrointestinal tract. The sustained inflammation is driven by the continuous influx of inflammatory leukocytes into the gut mucosa, which is regulated through chemokine gradients and adhesion molecules, including integrins. In the gut, the migration of inflammatory cells to specific segments is thought to be mediated by selected integrins and/or chemokine axes. The CCL25/CCR9 axis has been identified as a key pathway for immune cell migration into small bowel segments, such as the ileum. In this study, CITE-seq and spatial transcriptomic analysis was used to integrate proteomic and transcriptomic profiles to characterise CCR9+ immune cells in the ileum compared to the colon.

Methods
Biopsies were obtained via ileocolonoscopy from macroscopically inflamed and normal mucosa with appropriate consent from healthy volunteers and IBD patients visiting the Department of Gastroenterology at the Sahlgrenska Hospital. Biopsies were frozen in Cryostor CS10 or formalin fixed and paraffin embedded. Cells from frozen biopsies were sorted, CD45+ cells stained with CCR9 and Totalseq-C human universal antibody cocktail, followed by amplification, separation of antibody-derived DNA from mRNA derived cDNA, library construction and sequencing of both libraries. For spatial transcriptomics, tissue sections were deparaffinised, stained with H&E and decrosslinked before human whole transcriptome probe hybridisation, ligation, extension, library construction and sequencing. Cells were annotated using a transfer learning approach combining several reference single cell datasets including detailed CD4+ and CD8+ T cell datasets and utilising both antibody data and mRNA transcriptomic data.

Results
CCR9 positive cells appeared scattered throughout the lamina propria and expression at mRNA level was enriched in the ileum compared to the colon with about 10% of CD4 and 17% of CD8 T cells expressing CCR9. Interestingly at protein level, expression was more restricted with the highest expression on cells that exhibited proteomic and transcriptomic features associated with resident CD4 memory T cells (Trm). This population almost exclusively (&gt;90%) consisted of CCR9 positive cells and their proteomic and transcriptomic profile consistent with the recently described effector and innate-like phenotype of Trm cells that is thought to contribute to Crohn’s disease pathology.

Conclusion
These data may point to CCR9 positive cells, and in particular Trm cells, in the ileum as important pathological cells in Crohn’s disease and provide further support for depleting CCR9 cells as a potential therapeutic target for the treatment of ileal Crohn’s disease.
P0154 Adenosine A2B receptor signaling induces mucin expression in the intestinal epithelium
Authors: Jäverfelt S , O’Connell L V , Clare K , Aherne C M 
Published: January 2025

Abstract

Background
Loss of mucosal barrier integrity and inappropriate perpetuation of the inflammatory response are key features of ulcerative colitis (UC). Increased permeability and disruption of the mucus layer lining the colonic intestinal epithelium is observed prior to the onset of colitis in UC and murine colitis models. Therefore, understanding how the integrity of the mucus layer is maintained during health and disease may lead to novel therapeutic approaches for UC.
The major constituents of the mucus layer are mucins which are secreted by goblet cells in the intestinal epithelium. MUC2 is the major mucin within the ileum and colon, and MUC5AC has recently been demonstrated to be protective in murine colitis. Extracellular adenosine signaling can mediate mucin hypersecretion in asthma and other inflammatory respiratory conditions. Studies suggest that signaling via the Adora2b adenosine receptor (A2BAR) is protective in acute gastrointestinal inflammation such as colitis, although the mechanism by which this occurs is not known. We hypothesized that A2BAR signaling induces MUC2 and MUC5AC expression in the intestinal epithelium.

Methods
We are combining human epithelial cell lines and mouse models to investigate the A2B-mediated effects on MUC2 and MUC5AC in the intestine during homeostasis and injury. MUC2 and MUC5AC mRNA expression were quantified by Taqman RT-PCR and MUC5AC protein expression by immunofluorescent stainings. To investigate A2B-mediated effects on mucus thickness, growth and penetrability, we use an ex-vivo method where the intestinal tissue is mounted in a horizontal perfusion system equipped with temperature control. Mucus thickness, expansion and penetrability to bacteria-sized beads are recorded before and after basolateral and apical stimulation with a selective A2B during a total of 60 minutes.

Results
Treatment of human intestinal epithelial cells with an A2B selective receptor agonist induced upregulation of MUC2 and MUC5AC mRNA, which was abolished by receptor blockade and inhibition of PKA. Similarly, stimulation of the A2B-receptor with the selective A2B agonist increased MUC5AC promotor activity. Moreover, we assessed A2B-mediated induction of MUC2 and MUC5AC expression during epithelial injury. A2B agonist treatment of wounded intestinal epithelium resulted in increased MUC2 and MUC5AC mRNA expression and MUC5AC protein expression. We further explored the A2B-mediated effects on mucus growth and penetrability during homeostasis by an ex-vivo approach.

Conclusion
Our preliminary findings suggest that signaling through the A2BAR can increase MUC2 and MUC5AC expression in the intestine during homeostasis and injury.
P0155 Investigating the effect of a novel bio-based tissue glue for Perianal Fistulas in Crohn's Disease: A Low-Cost Rodent model study
Authors: Raghunathan N , Tokala R , Nagendra B , Pillikala M , Bera A K , Adhikari J , Reddy H , B Lakshmi J , Midde S , Singh J , Chavan S , Patel R , Sekaran A , Mitnala S , Pati F , Banerjee R 
Published: January 2025

Abstract

Background
Perianal fistulas (PAFs) are a severe complication of Crohn’s disease (CD) significantly affecting quality of life with limited treatment options. TNBS-induced colitis rodent models to study CD-PAF is costly and time-intensive. This study aimed to study the efficacy of a novel decellularized dermal extracellular matrix (ddECM) derived from goat skin as a tissue glue 1 for PAF treatment in a low-cost acetic-acid induced CD-PAF rodent model.

Methods
Two fistula tracts (0.3mm,1.3mm from the anal sphincter) were created in 12 male Wistar rats (aged 8-10weeks, body weight 180-220g); and a seton was placed to keep them open(Fig1A) 2. 500µL of 5%(v/v)acetic acid was injected biweekly intrarectally for a week to induce colitis and fistula discharge . Ultrasound (USG), in vivo imaging (IVIS) using indocyanine green fluorescent dye, and histopathology were used to assess inflammation and fistula formation(Fig2). 1ml bio-based tissue glue was applied to occlude each of the orifices and polymerized with UV light for 30-seconds. Rats were monitored for one week post glue treatment with single doze of 5% acetic acid as maintenance. USG was repeated to confirm healing. Upon sacrifice, tissue was collected for histopathology examination to confirm the success of CD-PAF and healing.

Results
All rats exhibited soft feces, diarrhoea, widening of fistula orifice, purulent fistula discharge, blood per rectum and significant weight loss after induction(Fig1B). USG confirmed active inflammation revealing edematous fistula tracts on both sides. This was validated by strong fluorescent signal in IVIS at the site of inflammation. Histopathology staining also showed acute inflammation (Fig2). One rat died post-induction. Tissue glue was applied to the remaining 11 rats. Upon one week follow-up, 10 of 11 rats exhibited normal feces, closure of fistula, absence of purulent fistula discharge and weight gain. USG and histopathology showed complete fistula healing in 10 rats and partial healing in one.

Conclusion
This study has demonstrated the use of low-cost CD-PAF rodent model which offers a faster induction (1-week) than conventional TNBS models (8 weeks). The novel bio-based tissue glue has shown to be a feasible and an effective treatment for CD-PAFs.

References
1. Bera, A.K. et al. BioScience.2022
2. Ling Li et al. Sci. Adv. 2023
3. Singh A. et al. Inflamm Bowel Dis. 2023
P0156 Plasma immune protein profiles in therapy-naïve ulcerative colitis relate to extent of affected tissue and density of intestinal infiltration, differentiating patients with discreet disease courses
Authors: Calado B , Heredia M , Charrout M , Klomberg R C W , Barendregt D , Hulleman-van Haaften D H , Tuk B , van Berkel L A , Folly B B , Escher J C , de Ridder L , Samsom J N 
Published: January 2025

Abstract

Background
Ulcerative colitis (UC) is a chronic intestinal inflammation driven by inflammatory memory T helper cell (ThM) reactivation. Heterogeneity in clinical disease and variability in response to treatment require new strategies targeting the patient’s underlying immune disease. As inflammation in UC only affects the intestinal mucosa, it is debated whether peripheral blood immune responses could reliably reflect ongoing intestinal immune disease.

Methods
To unequivocally establish this, we performed a uniquely deep immune characterization of therapy-naïve pediatric UC patients (n=34) and controls (n=55) by measuring 92 plasma immune proteins, circulating ThM subpopulations, intestinal mRNA expression, and intestinal immunohistochemistry. Extensive clinical follow-up was used to relate immune responses to clinical disease severity and therapy response.

Results
At diagnosis, 23 plasma proteins were differentially abundant in UC compared to controls. Increased protein concentrations mainly related to the IL-17 pathway (IL-17, IL-8, CCL20), tissue remodelling (MMP-10, MMP1, HGF), acute phase response (IL-6, OSM) and chemokinesis (CXCL9, CXCL10, CCL3). Interestingly, intestinal mRNA expression of highly abundant plasma proteins similarly increased, whereas no proportional changes in circulating ThM were detected. Unbiased hierarchical clustering of plasma immune profiles yielded two clusters of UC patients; UC_A with significantly increased concentrations of 17 proteins, many of which Th17/neutrophil-related, versus UC_B with lower concentrations. Notable increases in UC_A were: IL- 17A; OSM, a neutrophil product previously associated with anti-TNF therapy resistance; CXCL9 and CXCL11, IFN-g-induced chemokines. Strikingly, this UC_A immune profile strongly related to increased size of the affected intestinal surface and degree of infiltrating IL-17A+ cells and number of neutrophils in the biopsies. Clinically, UC_A patients had higher clinico-pathological parameters, more severe intestinal disease at endoscopy and on histology , and required earlier treatment intensification during 1.5 year follow-up, compared to UC_B patients.

Conclusion
These data firmly establish that, even though inflammation in UC only affects the intestinal mucosa, peripheral blood immune protein responses reliably reflect the degree of ongoing intestinal immune disease in UC. Implementing immune profiling in routine clinical characterization at diagnosis improves identification of patients who will develop a severe disease course allowing tailored treatment with earlier start of more intensive immunomodulatory treatment to treat to target and avoid tissue damage.
P0157 Preoperative emotional dysfunction is associated with postoperative morbidity in Crohn’s disease patients
Authors: Schmitz A , Bercher M , Iesalnieks I 
Published: January 2025

Abstract

Background
Anxiety has been found to be associated with postoperative morbidity in patients with Crohn’s disease. The impact of preoperative emotional dysfunction was analyzed in the present prospective observational study.

Methods
Consecutive patients undergoing bowel resection for Crohn’s disease were asked to fill out the IBDQ-32 questionnaire. The 32-item Inflammatory Bowel Disease Questionnaire (IBDQ-32) is the most frequently used instrument to capture disease- specific quality of life. One of 4 domains scores the emotional function. The domain consists of 12 items. All items use 7-point Likert-type scales for capturing symptom- related experiences over the previous 2 weeks, with 1 indicating the highest symptom frequency/severity and 7 indicating the lowest symptom frequency/severity. For emotional function, scores between 12 and 84 can be calculated, higher score indicating better quality of life.

Results
112 patients underwent bowel resection (90 ileocolic resections, 11 colorectal resections, 5 both ileocolic and colorectal resections and 6 small bowel resections. Postoperative complications occurred in 26 patients (23%), postoperative ileus in 10 (9%), wound complications in 9 (8%) and intraabdominal septic complications in 5 (4.5%). Patients who developed postoperative complications revealed significantly decreased emotional scoring (40.3 vs. 48.3, p=0.009). Also, patients with postoperative ileus (38.2 vs. 47.1, p=0.049), wound complications (35.1 vs. 47.4, p=0.009) and intraabdominal septic complications (40.8 vs. 46.6, p=0.35) demonstrated higher grade of emotional dysfunction preoperatively. Preoperative abscess, severe preoperative anemia and preoperative weight loss were all significantly associated with an impaired preoperative emotional functioning.

Conclusion
Preoperative emotional dysfunction was associated with an increased risk of postoperative morbidity. However, severe disease complications (abscess, anemia,weight loss) led also to more emotional suffering preoperatively.
P0161 The primary sclerosing cholangitis and ulcerative colitis colonic mucosa defined through paired microbial and single-cell RNA sequencing
Authors: Tearle J , Zhang F , Jackson K , Malhotra P , Tavakoli P , Koentgen S , Warren J , Williams C , Haque A , Arzivian A , Tedla N , Kim A , King H , Hold G , Ghaly S , James K 
Published: January 2025

Abstract

Background
Primary sclerosing cholangitis (PSC) is a chronic progressing cholestatic disease that often co-occurs with inflammatory bowel disease (PSC-IBD). PSC-IBD affecting the colon (PSC-UC) is likened clinically to ulcerative colitis (UC), however differences include a right colon dominance, less severe inflammatory presentation and a greater lifetime risk of colorectal cancer.

Methods
We combine single-cell mRNA and antigen receptor sequencing, 16S ribosomal RNA gene analysis and spatial transcriptomics to profile mucosal biopsies from four colon regions of PSC-UC and UC patients in remission or at the time of relapse.

Results
In contrast to UC, PSC-UC patients display an enrichment of CD8 T, γδ T and natural killer cells and reduced microbial diversity in the right colon, even in the absence of symptomatic inflammation. During flare, we demonstrate the expansion of a novel inflammatory mast cell state in both diseases and highlight the function of TMEM176B in sustaining this activated state.

Conclusion
These results highlight that the PSC-UC and UC colonic mucosa are fundamentally different with shared cell programs during active disease, providing insights to guide tailored clinical management and precision therapies.
P0163 Novel IBD treatment: Small molecule oral GPRx agonists for Treg cell therapy
Authors: Zhou K , Shi Y , Wang T , Fan F , Xia T 
Published: January 2025

Abstract

Background
Our study has revealed that GPRx regulates gut DC migration to local draining lymph node to induce iTreg (Fig 1).1 To fulfil our drive to drug discovery and development for IBD (and other autoimmune diseases) program, we’ve rationally designed small molecule oral GPRx agonists that can induce gut antigen specific iTreg and intended to examine their efficacy utilizing TNBS induced acute IBD model.2-4

Methods
C57/BL6 8-10 weeks old female mouse was intrarectally administered with 2,4,6-trinitrobenzene sulfonic acid (TNBS) in 30% ethanol solution through a special catheter to induce colitis, from day 0 to day 4. The tested drug was given via PO, from day 0 to day 4. The experiment ended on day 5. DAI (disease activity index) on weight loss, diarrhea, and rectal bleeding, was recorded daily, to determine disease severity. Colon was taken at the end of experiment for measurement and photography.

Results
A series of compounds have been tested using this in vivo TNBS induced acute IBD mouse model and some of them have shown excellent activity (Fig 2).

Conclusion
With this new MOA and first-in-class approach using small molecule oral GPRx agonists for Treg cell therapy, we could develop a different treatment for IBD (and other autoimmune diseases in general) patients than JAK inhibition or a4b7 inhibition or S1P1 activation, etc. The outstanding efficacy data described here together with their safety profile and acute/sub-acute toxicity results have provided us the confidence to a successful IND late-2025.

References
1. Unpublished data
2. ZL2017107741357
3. PCT/CN2023/135952
4. PCT/CN2023/135912
P0166 Changes in bone mineral density in young patients with inflammatory bowel diseases
Authors: Myasoutova E , Shamsutdinova N , Abdulganieva D 
Published: January 2025

Abstract

Background
Inflammatory bowel diseases (IBD), which include Crohn’s disease (CD) and ulcerative colitis (UC), are an urgent problem of modern gastroenterology, which is associated with an increase in morbidity, a lifelong recurrent course, and the prevalence of the disease mainly among young people of working age. Extracellular manifestations in CD and UC include damage to the bone system in the form of impaired bone mineralization with the possible development of osteoporosis (OP). In this regard, patients with IBD have a high risk of developing osteoporotic fractures.

Methods
The aim of the study was to evaluate the absolute values of bone mineral density (BMD) in young patients with IBD.
96 patients with a verified diagnosis of IBD were examined: 41 patients had CD, 55 had UC; men -50, women - 46. The average age of the patients was 29 [25;35] years. The average age of IBD debut was 24 [19.5;29] years. All patients underwent a general clinical examination, as well as a two-adsorption X-ray densitometric examination of the lumbar spine and proximal femur on the right and left with the determination of BMD and bone mineral content (BMC). The data obtained were correlated with the NHANES III database.

Results
The average index of the Z-criterion (L1-L4) was -0.4 [-1.4;0.4]. The average Z index (hip neck) was -0.4 [-1.2;0.5]. The average BMD value at the level of the lumbar spine (L1-L4) was 1.175 [1.07;1.28] (g/cm2). The average total BMC was 53.49 [45.83;60.04] g. The average BMD value at the level of the neck of the right femur was 0.99 [0.88;1.09] (g/cm2). The average value of the total BMC in the same place was 24.86 [22.35;28.58] g. The average BMD value at the level of the neck of the left femur was 0.97 [0.88;1.09]. The average total BMC value was 25.75 [22.34;28.82]. 13.5% (13 out of 96 patients) showed a decrease in bone mass below the chronological age.

Conclusion
Despite the young age of the patient, a significant proportion of patients (13.5%) have deviations in BMD from normal values. Also, the absolute values of BMD and bone mineral content differ from the values when compared with the reference interval in the NHANES III database.

References
1.Kreienbuehl A. S. et al. Bone health in patients with inflammatory bowel disease //Swiss medical weekly. – 2024. – Т. 154. – №. 3407. DOI: 10.57187/s.3407
2.A.Kh. RAMAZANOVA, I.G. MUSTAFIN, A.Kh. ODINTSOVA, R.M. NABIULLINA, D.T. SIRAZETDINOV, D.I. ABDULGANIEVA Inflammatory bowel diseases: epidemiology and development of thromboembolic complications, Practical medicine №4 (96) 08/2016/volume 2; 96: 88-91
P0174 Enteric glial cells-derived NGF prevents necroptosis of intestinal epithelial cells and alleviates DSS-induced colitis
Authors: Min Z , Zhang Y , Hu C , Song X , Guo H 
Published: January 2025

Abstract

Background
The maintenance and repair of the intestinal epithelial barrier (IEB) is a multifaceted process that requires the coordinated participation of different cell types within the gastrointestinal tract. Studies have shown that ablation of enteric glial cells (EGCs) can lead to intestinal epithelial cell necrosis and destruction of the IEB. However, the specific role of EGCs in it remains ambiguous. The aim of this study was to elucidate the potential role of EGCs in regulating IEB and intestinal inflammation.

Methods
In this study，we employed immunofluorescence, western blot, ELISA, and gene expression profiling to assess the expression of GFAP (a marker for EGCs), NGF, and necroptosis-associated proteins in intestinal tissues from IBD patients. In addition, DSS-induced colitis mouse models, in vitro co-culture systems, and transcriptomic analysis were used to explore the effects of EGCs and NGF on IECs necroptosis and inflammation.

Results
In intestinal tissues of IBD patients, reduced expression of GFAP and S100β suggests damage to EGCs. Additionally, we found that NGF in intestinal tissues primarily originates from EGCs. Diminished NGF secretion due to EGCs damage correlates with increased necroptosis of IECs. Exogenous administration of NGF inhibits DSS-induced necroptosis of IECs and alleviates colonic inflammation. Ablation of EGCs exacerbates necroptosis of IECs, disruption of IEB, and intestinal inflammation following DSS treatment. In vitro co-culture experiments demonstrate that EGCs suppress necroptosis of IECs induced by T/S/Z mix. Further flow cytometry and TEM results confirm that NGF effectively mitigates T/S/Z mix-induced necroptosis of IECs.Transcriptomic sequencing indicated that RNF126 is a downstream target in the NGF signaling pathway, potentially mediating the protective effects of EGCs-derived NGF on IECs.

Conclusion
Based on the findings from this study, it is evident that EGCs play a crucial role in maintaining intestinal homeostasis through their production of NGF. Reduced NGF secretion from damaged EGCs correlates with increased necroptosis of IECs, exacerbating inflammation in colonic tissues. Exogenous NGF administration effectively attenuates this process, highlighting its therapeutic potential in IBD. Furthermore, transcriptomic analysis identified RNF126 as a downstream mediator in the NGF signaling pathway, its specific role remains speculative and requires further investigation to elucidate its mechanistic involvement in mediating the protective effects of NGF. In conclusion, these findings emphasize the intricate interplay between EGCs, NGF signaling, and IEC survival, pointing towards novel therapeutic strategies targeting this pathway in IBD management.