bluuebunny/crossref_metadata_2025_split huggingface dataset:
P014 Behaviour in mice with chronic DSS colitis mimics fatigue in IBD and is associated with neuroinflammation
Authors: Truyens M , Lernout H , Vandendriessche C , Bruggeman A , Xie J , De Vos M , Vermeirssen V , Vandenbroucke R , Laukens D 
Published: January 2023

Abstract

Background
Inflammatory bowel diseases (IBD) are often associated with psychological comorbidities such as fatigue. Up to 47% of patients reports fatigue despite disease remission. The aim of the current study was to assess neurobehavioural changes in a mouse model of extinguished chronic colitis and to explore associated changes along the gut-brain axis.

Methods
Repeated administration of 2% dextran sulphate sodium (DSS) was used to induce chronic colitis in C57BL/6 mice, followed by a recovery period of 3 weeks to mimic quiescent IBD (Figure 1). Behavioural testing was performed at baseline and after the recovery period and compared with control mice. RNA sequencing was performed of the distal colon and choroid plexus and weighted correlation network analysis (WGCNA) was applied. Next, quantitative polymerase chain reaction (qPCR) was used for the confirmation of the expression levels of genes identified in the WGCNA analysis. The number of microglia and their morphology were assessed by Iba1 immunofluorescence in a second experiment, also including acute colitis (aDSS), which was induced by 1 week of 2% DSS, with sampling at day 7.

Results
Chronic DSS (cDSS) treatment resulted in chronic inflammation with minimal residual intestinal symptoms (control: median disease activity index (DAI) of 0 IQR [0-0], cDSS: median DAI of 0.5 [0.5-0.88], P &lt;0.0001) after 3 weeks of recovery, which mimics quiescent IBD. At the time of sampling, the mean weight and haematological parameters (including red and white blood cells) in cDSS were comparable to those in control mice. cDSS mice exhibited significantly reduced spontaneous activity compared with their baseline behaviour (P = 0.0084) and with the controls (P = 0.0186). WGCNA on the colon and the choroid plexus expression data revealed a consensus module of 123 co-expressed genes that differed significantly between the control and cDSS mice (P = 0.00001721). These genes were implied in pathways of neutrophil and complement activation and qPCR on different brain regions (frontal cortex, hippocampus and choroid plexus) confirmed upregulation of inflammatory genes Lcn2, TNFa, S100A8 and S100A9 (Figure 2). Acute colitis was associated with activation of microglia in the prefrontal cortex and hippocampus, persisting in the prefrontal cortex of mice that recovered from chronic colitis (Figures 3 and 4).

Conclusion
In this mouse model of chronic extinguished colitis reduced spontaneous activity was seen, indicating fatigue-like behaviour as observed in patients with IBD. Moreover, mice that recovered from colitis exhibited persisting neuroinflammation.
P015 CITE-seq analysis revealed immune cell diversity in colonic mesenteric lymph nodes from IBD patients
Authors: Wils P , Mehta H , rubio M , Sarfati M , Chapuy L 
Published: January 2023

Abstract

Background
Mononuclear phagocytes (MNPs), including dendritic cells (DCs), monocytes (Mo), and macrophages (Mac), are key inducers of the adaptive immune response. MNPs could circulate between blood, mucosa, and mesenteric lymph nodes (MLNs), where they interact with T cells, natural killer (NK) cells, and innate lymphoid cells (ILC), contributing to chronic inflammation. MNPs have been previously investigated at the transcriptomic level in the inflamed gut mucosa, but their molecular characterization at the single-cell level remains uncovered in inflamed MLNs of patients with inflammatory bowel diseases (IBD).

Methods
MLNs were collected from surgical colonic resections of patients with Crohn’s disease (CD; n=3) or ulcerative colitis (UC; n=3). Cell suspensions were multiplexed and then pooled for sorting CD45+CD3+CD19- T cells and CD45+CD3-CD19- non-T/non-B cells by flow cytometry. Next, sorted cells were combined and labeled with BD Abseq antibodies (antibodies conjugated to oligonucleotide tag). CITE-seq (Cellular Indexing of Transcriptomes and Epitopes by Sequencing), combining simultaneous RNA and protein expression, was performed on the BD RhapsodyTM platform.

Results
High-quality data from 3308 captured single cells (1471 from CD and 1837 from UC) identified four main immune cell populations (myeloid DCs/Mo/Mac, plasmacytoid DCs, T cells, and NK/innate lymphoid cells). We further segregated cell subsets into 16 MNP, 4 NK/ILC, and 9 T cell clusters. MNP subsets were characterized according to specific gene expression (HLA-DRA, CD1c, ITGAX, SIRPA, JCHAIN, IL3RA). They revealed transcriptionally distinct DC clusters, including one cluster of mature regulatory DC (mregDC) (LAMP3, CCR7), one cluster expressing AXL and THBD, and one cluster of CD103+ DCs expressing LTB, FLT3, and IL22RA2. Further analysis of MNP subsets also revealed 6 clusters of monocytes and macrophages, including inflammatory Mo, Inflammatory Mac, and regulatory Mac. Overall, the proportion of myeloid cells and pDC was higher in UC MLNs when compared to CD (29.6% and 21.6% versus 17.1% and 11.7%, p&lt;0.0001) while the proportion of NK/ILC was higher in CD MLNs (11.7% versus 24.9%, p&lt;0.0001). Comparison of differentially expressed genes (fold-change&gt;1.5) between CD and UC samples showed that CD MLNs were enriched in NK-specific marker genes (KLRD1, IL2RB, NKG7, GZMA). In UC, the majority of upregulated genes related to myeloid gene markers (THBS1, HLA-DQA1, HLA-DRA, VCAN, IL1B, JCHAIN, and CXCL8).

Conclusion
CITE-Seq analysis of the colonic MLNs of patients with active IBD suggests differences in immune cell landscape gene expression between CD and UC.
P016 Dietary supplementation with Brazilian jaboticaba-sabará fruit (Myrciaria jaboticaba (Vell.) O. Berg) ameliorates colitis-driven colorectal cancer in mice
Authors: Nascimento R D P D , Rizzato J S , Polezi G , Moya A M T M , Silva M F , Franchi Junior G C , Borguini R G , Santiago M C P D A , Paiotti A P R , Pereira J A , Martinez C A R , Marostica Junior M R 
Published: January 2023

Abstract

Background
Individuals with inflammatory bowel diseases (IBD) are at a high risk of developing colorectal cancer (CRC). Recently, pre-clinical studies have demonstrated that South American crops are rich sources of bioactive compounds that can act against both IBD and CRC. Therefore, our study aimed to characterize the freeze-dried peel of jaboticaba-sabará (Myrciaria jaboticaba (Vell.) O. Berg), a native Brazilian berry, and understand its preventive effects on a mouse model of IBD-related CRC.

Methods
Jaboticaba peel powder was chemically characterized by its contents of polyphenols, carotenoids, and dietary fibers. For the animal experimentation, male BALB/c mice were given in their diets 0, 2.5, or 5% jaboticaba powder for 17 weeks. On day 28, animals received an injection of the carcinogenic azoxymethane. During days 35-42 and 49-56, a solution with dextran sodium sulfate was given as drinking water to mice to induce chronic colitis. Clinical parameters were checked frequently in order to compose the Disease Activity Index (DAI). On day 119, mice were euthanized, their organs were measured, and the colon was checked for visible tumors. Histology processing was performed to assess the incidence of adenocarcinomas and for a score analysis of inflammatory-related parameters.

Results
High-performance liquid chromatography showed that dry jaboticaba is a rich source of cyanidin-3-O-glucoside (1.45%), gallic acid (0.32%), ellagic acid (0.3%), and delphinidin-3-O-glucoside (0.1%). Other compounds were found in a lesser amount, including protocatechuic acid (0.012%), rutin (0.006%), syringic acid (0.005%), and lutein (0.002%). Additionally, the powder showed relevant contents of soluble (11.3%) and insoluble (24.9%) dietary fibers (Table 1). Animals with colitis-driven CRC had diarrhea and showed bleeding in their feces, besides an average of 11 ± 2.1 visible tumors. On the contrary, animals that received 5% jaboticaba had a significant recovery in their colon length and showed reduced signs of colitis, including less bloody diarrhea, therefore, reduced DAI. The 5% intervention decreased the number of tumors to an average of 4.2 ± 1.0 (Figure 1). Adenocarcinoma incidence went from 66.6% in the control cancer group to 37.5% in the 5% jaboticaba group, the latter in which 50% of the animals did not show any alteration (adenoma or adenocarcinoma). Finally, the 5% jaboticaba diet significantly reduced crypt distortion and the depletion of goblet cells. The 2.5% jaboticaba diet had no beneficial effects.

Conclusion
Our preliminary results indicate that anthocyanin- and fiber-rich jaboticaba could represent a potential prevention tool for IBD-related CRC. Further analyses are necessary to understand the molecular mechanisms of jaboticaba on CRC.
P021 Microbial metabolite receptors as mediators of pathogenic processes in Inflammatory Bowel Disease.
Authors: Spathakis M , Dovrolis N , Tarapatzi G , Filidou E , Kandilogiannakis L , Valatas V , Drygiannakis I , Manolopoulos V G , Kolios G , Arvanitidis K , Vradelis S 
Published: January 2023

Abstract

Background
Inflammatory Bowel Disease (IBD) is a chronic inflammatory condition of the alimentary tract, primarily manifested as Crohn’s disease and Ulcerative colitis. Dysbiosis of the intestinal microbial microflora is a well-established characteristic of IBD. The microbiota metabolism ends in the production of several metabolites that participate in interactions with the host and ultimately influence its physiology. Our aim is to investigate whether metabolite receptor expression differs between IBD and healthy individuals and unravel their possible interactions with inflammatory and fibrotic pathways in the intestine.

Methods
RNA-sequencing data from over 2500 intestinal biopsy samples were collected from publicly available datasets via the Sequence Read Archive. Bioinformatics analysis of metabolite receptor genes was performed using the RaNA-seq online platform which included normalization, alignment and differential expression. In addition, correlation analysis between the target receptor genes and inflammatory/fibrotic genes as well as functional analysis was performed via a R based pipeline to elucidate molecular interactions during IBD.

Results
The expression of 22 receptors responding to the major metabolite ligand classes bile acids, short chain fatty acids and indole derivatives from tryptophan metabolism was targeted. Several receptors of bile acids as well as receptors of short chain fatty acids, such as the vitamin D receptor (VDR), the peroxisome proliferator-activated receptor alpha (PPARα) and the hydroxycarboxylic acid receptor 2 (HCAR2), were found differentially expressed in intestinal biopsies of CD patients as compared to healthy individuals. Negative and positive correlation of these receptors with genes of known involvement in IBD was explored, along with their simultaneous contributions to inflammatory and fibrotic processes.

Conclusion
Our results demonstrate the interplay between the intestinal microbiota and the host on a metabolite-receptor level and identify disease related interactions with inflammation and fibrosis.
P023 Immune-metabolic profiles of creeping fat in relation to degree of severity in Crohn’s disease postsurgical recurrence.
Authors: Suau R , Pardina E , López-Tejero M D , Lorén V , Zabana Y , Llaves M , Cordobilla B , Cañete F , Guasch M , Ricart-Jané D , Mañosa M , Domènech E , Domingo J C , Manyé J 
Published: January 2023

Abstract

Background
Crohn’s disease (CD) is an inflammatory bowel disease (IBD) of complex etiology that normally requires bowel resection. However, many patients suffer from post-operative recurrence (POR) within one year. Here, the inflammatory and bacterial influence of the affected intestine transforms the neighboring mesenteric tissue to creeping fat (CrF). The main objective has been to characterize the immune-metabolic profile of CrF according to the POR severity and compare them with mesenteric tissues from IBD-free controls (colorectal cancer and severe obesity).

Methods
POR severity was classified using the Rutgeerts index: non-recurrents (i0+i1, n=18; NR), mild-recurrents (i2, n=14; MR) and severe-recurrents (i3+i4, n=10; SR). The CrF were gathered from intestinal resections, and IBD-free mesenteric tissues were collected from intestinal resections for colorectal cancer (n=10) and bariatric surgery for severe obese patients (n=11). The total DNA was quantified using DABA assay, 105 cytokines were semiquantified by immune blotting and the fatty acid (FA) profile was quantified by GC-MS.

Results
The DNA analyses in the CrF showed a major cellularity demonstrating the hyperplasia. When compared to the control groups and to SR, the NR patients showed an increment of infection pro-resolutive cytokines (IFN-γ, MIF, BAFF), and, specially, an increase in both Th1 and Th2 cytokines (IL-18, IL-33). Here, there was also an increase in molecules related to angiogenesis (endoglin) and cellular adhesion (ICAM-1 and VCAM-1). On the other hand, the SR showed a higher level of pro-fibrotic PAI-1 and of the leukocyte homing inducer CCL5.

Going into the FA, in comparison to the control groups, there was an increase in the saturated in front of a decrease in mono-unsaturated FA in all the CD groups. Regarding the polyunsaturated FAs, the CrF and the obese mesentery showed a general increase of the omega 6/omega 3 FA ratio when compared to colorectal cancer controls.

Finally, both obese and NR CD patients had less short-chain FAs, specifically acetate and propionate, than the colorectal controls. Unexpectedly, the butyrate levels were very low in all the mesenteric tissues.

Conclusion
The CrF from NR CD patients show an infection resolutive profile regarding the controls and the SR. Instead, these lasts showed signs of chronic inflammation and a fibrotic profile. In general, CrF FA proportions suggests changes in cell signaling due to loss of cell membrane fluidity. In addition, the greater presence of omega-6 than omega-3 FA in the CrF and mesentery of obese patients could be related to the generation of potent proinflammatory lipid mediators. In contrast, the propionate and acetate decrease detected in NR CD patients could be protective against POR.
P024 Effects of anti-TNF and anti-interleukins 12 and 23 drugs on circulating dendritic cells migratory capacity in inflammatory bowel disease
Authors: Soleto I , Ramirez C , Gómez C , Montse B , García S , Orejudo M , Mecado J , Chaparro M , Gisbert J P 
Published: January 2023

Abstract

Background
Inflammatory bowel disease (IBD) is an idiopathic and chronic disorder that includes ulcerative colitis (UC) and Crohn’s disease (CD). Both diseases are different but show an uncontrolled intestinal immune response that generates tissue inflammation. Dendritic cells (DC) are phagocytic professional antigen presenting cells that link the innate and adaptive immune systems. Indeed, they are the only cell type able to stimulate naïve T cells generating an antigen-specific immune response. In humans, DC can be divided into plasmacytoid (pDC) and c conventional type 1 and 2 (cDC1, cDC2). Although cDC migration towards the GI mucosa is enhanced in IBD, the mechanisms underlying these migrations and whether biological drugs can modulate them are currently unknown. This study aimed to analyze DC migratory capacity towards the GI mucosa, and whether biological drugs with different mechanisms of action can modulate this process in health and IBD.

Methods
This is a cross-sectional observational postmarketing study where the modulatory effect of different biological treatments on samples from patients with an endoscopic diagnosis of IBD and healthy controls was studied ex vivo. Peripheral blood mononuclear cells (PBMC) were obtained from 15 patients per group and were cultured in presence of golimumab or ustekinumab. Following culture, surface markers expression was determined, assessing the differential effect of biological drugs (if any) over the DC subsets in the different groups. After that, the migratory capacity of circulating DC towards CCL2, CCL25, and MadCam1 from the different study groups was determined. Finally, we evaluated the DC migratory capacities in presence of biological drugs or with culture media alone.

Results

The cCD2 subset showed higher expression levels of β7 and CCR5 compared to cCD1 (Figure 1). Golimumab and ustekinumab did not modify the expression of the homing markers in any studied subset (Figure 2). Even though no conclusive results were obtained for the biological drugs used in the study on the expression of homing markers, ustekinumab decreased the migratory capacity of cDC2 from CD patients towards a medium supplemented with CCL25 (Figure 3).

Conclusion
cDC2 showed more migratory profile markers than other DC populations. Golimumab and ustekinumab did not modify the expression of the homing markers in any DC subset in any group. Nevertheless, ustekinumab decreased the migratory capacity of cDC2 from CD patients. This fact indicates that the behavior of this population is different in CD and UC and that the migration capacity of this subset is not dependent on homing markers.
P025 Colonic rather than ileal expression of ITGA4/ITGB7 correlates with local intestinal inflammation in IBD
Authors: Csomor E , Gehrmann U , Monkley S , Tian S , Cairns J , Angermann B , Khan E , Nys J , Marks D 
Published: January 2023

Abstract

Background
Lymphocyte trafficking to the inflamed gut is one of the hallmarks of Inflammatory Bowel Disease (IBD) pathology. Integrin a4b7 and its ligand MADCAM1 play an important role in targeting immune cells in the bloodstream to mucosal tissues and blocking this pathway using the anti-a4b7 monoclonal antibody Vedolizumab (VDZ) proved to be an effective treatment in IBD. However, VDZ is most efficient in treatment of Ulcerative Colitis (UC) and of colonic rather than ileal Crohn’s Disease (CD), for which the reasons have not been completely understood. Here we sought to compare the expression of ITGA4, ITGB7 and MADCAM1 in gut tissue biopsies from small and large bowel with respect to the underlying severity of tissue inflammation.

Methods
Differential gene expression analysis was conducted using bulk RNA Sequencing data generated from gut tissue biopsies of the large and small intestine of 611 CD and 315 UC patients enrolled in a multicentred longitudinal Study of a Prospective Adult Research Cohort with IBD (SPARC IBD) obtained from the IBD Plexus program of the Crohn’s and Colitis Foundation. Gene expression levels of ITGA4, ITGB7 and MADCAM1 were compared across ileal, colonic and rectal segments of the intestine and data were then linked to available measures of local inflammation.

Results
ITGA4, ITGB7 and MADCAM1 were expressed at comparable levels in large vs small bowel biopsies. Expression of MADCAM1 was significantly increased in both ileal and colonic biopsies from patients with moderate to severe CD based on segmental or composite endoscopy score (SES-CD) across all bowel segments. Expression of both ITGA4 and ITGB7 was similarly increased in colonic biopsies of moderate/severe CD patients, whereas only minor changes were observed for both genes in ileal biopsies. Similar findings were obtained from UC patients, where expression of MADCAM1 was significantly increased in inflamed or ulcerated biopsies from both sigmoid colon and cecum as compared to normal appearing biopsies. Expression of ITGA4 and ITGB7 was only increased in inflamed/ulcerated biopsies of the sigmoid colon but not the cecum and correlated with the modified Mayo endoscopic score (MMES).

Conclusion
Our analysis suggests that influx of leukocytes to inflamed tissue via ITGA4/ITGB7 and MADCAM1 interactions is more common in the distal colon compared to proximal colon or ileum. This provides a possible mechanistic explanation as to why VDZ is most effective in patients with colonic IBD. Further work is needed to understand the differential regulation of cell trafficking to different regions of the gut.
P035 Anti-inflammatory Effect and Mechanism of Action of BMC333, a Rationally-Designed Live Bacterial Consortium Based on Microbiome Functional Genomic Analysis for Treating IBD
Authors: Tirosh O , Meshner S , Eshar S , Kruger Ben-Shabat S , Polonski O , Haber E , Liu B , Herzog J , Sartor R B , Ringel Y 
Published: January 2023

Abstract

Background
The mechanisms by which the intestinal microbiome drives inflammation and disease activity in IBD is not clear. Most of the current efforts in developing treatments targeting the intestinal microbiome in IBD are focused on microbial taxa and very few treatments are developed based on microbiome functions in this condition. Using high-resolution functional microbiome analysis we have designed a live bacterial consortium consisting of four bacterial strains with anti-inflammatory functions (BMC333); and demonstrated the significant anti-inflammatory effects of this consortium in a mouse model of DSS-induced colitis by improving disease activity index, colon length, histologic score and fecal lipocalin (presented at DDW 2022).

Aim
To validate the anti-inflammatory effects and mechanisms of action of BMC333 using IL10-reporter mouse model.

Methods
Germ free mice bearing the IL10-GFP reporter gene (B6. IL10-GFP) were orally gavaged with BMC333 at 109 CFU/strain or vehicle/placebo (n=6 in each group) at experimental days 1 and 4. The induction period was followed by 10 days without intervention and the animals were sacrificed at day 14. Immunologic effects was assessed in mouse splenocytes and lamina propria by I. quantitating lamina propria immune-suppressive cells using flow cytometry. II. Comparing the effect of stimulation with each of BMC333 bacterial strains’ lysates on IL10- and IFNγ production by splenocytes from BMC333 or vehicle-treated mice.

Results
Treatment with BMC333 resulted in:
I. Higher levels of immune-suppressive cells (IL10-expressing CD4 T cells and B cells, and regulatory T-cells) in the lamina propria of mice treated with BMC333 compared with vehicle-treated mice.
II. Higher numbers of IL10-expressing splenic immunocytes stimulated with lysates compared to non-stimulated cells. Additionally, splenocytes stimulated by each of the BMC333 bacterial strains secreted higher IL10 levels, whereas the strain lysates did not induce secretion of the pro-inflammatory cytokine IFNγ, as measured by ELISA.

Conclusion
These results validate the anti-inflammatory effects of BMC333 and indicate that this beneficial effect is, at least partially, mediated by increasing IL-10 anti-inflammatory activity as demonstrated by the increased numbers of IL-10-producing lamina propria T and B cells after in vivo colonization, and higher IL-10 producing cells and secretion by BMC333 lysate-stimulated splenocytes. BMC333 stimulates increased immuno-suppressive cell populations in the lamina propria, but no increased production of the pro-inflammatory cytokine IFNγ. These findings are consistent with our previously reported in vivo studies and provide a strong rationale for further development of BMC333 as an effective treatment for IBD.