bluuebunny/crossref_metadata_2025 huggingface dataset:
P019 Development of hydrogel systems for targeted rectal delivery of intestinal cells to injured mucosa in inflammatory bowel disease
Authors: Mantaj J
Published: January 2024

Background

Crohn’s Disease (CD) and Ulcerative Colitis (UC) are Inflammatory Bowel Diseases (IBD) affecting over 6.8 million people worldwide. CD and UC are chronic and progressive diseases, characterised by destructive inflammation of the intestinal tract. IBD management consists of drugs such as steroids, aminosalicylates, immunosuppressants, and biologics that aim to induce disease maintenance and remission. However, frequent, and systemic drug administration leads to limited local effects, severe side effects, and low patient compliance. Ultimately, about 20% of UC and up to 80% of CD patients require surgical colon removal. Thus, new approaches to locally restore the inflamed mucosa and initiate colonic wound repair are required.

Human intestinal organoids (HIOs) are 3D cell aggregates derived from primary tissue or stem cells grown in vitro. They demonstrate a similar composition architecture to the primary intestinal tissue. Studies showed that HIOs can engraft into injured colonic mucosa resulting in wound repair. Thus, delivery of HIOs to damaged intestinal epithelium represents a promising therapeutic option in IBD treatment.

The project aims to develop hydrogel systems with embedded HIOs and obtain human-relevant information on their efficacy in the restitution of injured colonic mucosa using an improved and human-relevant organ-on-a-chip technology.

Methods

A series of thermoresponsive hydrogels were fabricated (based on methylcellulose/hyaluronic acid polymers) and characterised. Following the embedment of HIOs into hydrogels, the morphology and viability of HIOs were determined using microscopy and MTT assays. Inflammation was induced through the administration of LPS to the basolateral side of the gut chip. Hydrogel/HIO systems were applied to the apical chip compartment via flow for 24 hours. Subsequently, hydrogel/HIO systems were removed from the chip, and HIO engraftment into injured mucosa was determined utilising permeability (4 kDa dextran) and cytokine release assays (ELISA IL-8).

Results

Our findings show that the gels were liquid at 25 °C and solidified at 37 °C and demonstrated low cytotoxicity. Additionally, our results suggest that the hydrogel systems had a restorative and possibly protective effect on the villi structures in the gut chip. Microscopy results clearly demonstrated the presence of fully differentiated villi structures in the chips that were treated with the hydrogel/cell systems (Figure 1B). These villi structures were absent in the not-treated chips (Figure 1A).

Conclusion

This study suggests that development of organoid-based hydrogels that can serve as a safe, effective, and inexpensive therapy (compared to surgery) and significantly improve the treatment of IBD.
P024 Synergistic effect of phytochemicals combination including ginsenosides and curcumin on recovery from radiation-induced toxicity
Authors: Kim S E , Kim M S , Lee R A , Yang S J , Lim H , Jung S Y , Lee T Y , Park J K , Park Y G , Woo S Y
Published: January 2024

Background

Radiotherapy is commonly used to treat solid cancers located in the pelvis. A considerable number of patients experience proctitis of varying severity, even for a considerable period after radiotherapy. These side effects are often long-lasting or progressively worsen despite multiple therapeutic efforts and are a primary cause of an unexpectedly low quality of life, even after successful cancer treatment. Therefore, this in vitro study, as an initial trial, evaluated the single and combined efficacy of ginsenoside, curcumin, butyric acid, and sucralfate compounds in treating radiation-induced proctitis.

Methods

Cell viability and migration assay, endothelial cell tube formation assay were performed using human colon cancer cell lines, dermal fibroblasts, keratinocytes, and human umbilical vein endothelial cells, which were exposed to radiation. In addition, Inflammatory cytokine assay for CXCL10, CCL2, CCL8, IFN-γ, IL-4, IL-12, and IL-21 is performed using human monocytes treated with LPS (lipopolysaccharide). In all assay, the single and combined efficacy of ginsenoside, curcumin, butyric acid, and sucralfate were analyzed.

Results

While the candidate compounds did not affect the proliferation and migration of cancer cells, they promoted the recovery of cell activity, including motility. They exhibited anti-inflammatory effects on human dermal fibroblasts or human umbilical vein endothelial cells within in vitro disease models. When each compound was tested, curcumin and ginsenoside were the most effective in cell recovery and promoted the migration of human dermal fibroblasts and cell restoration of human umbilical vein endothelial cells. The combination of ginsenoside and curcumin resulted in cell migration recovery of approximately 54%. In addition, there was a significant improvement in the length of the endothelial tube, with an increase of approximately 25%, suggesting that the ginsenoside-curcumin-containing combination was the most effective against radiation-induced damage. Furthermore, it was shown that the combination could alleviate radiation-induced inflammation by reducing the concentrations of IFN-γ and IL-12, associated with activated macrophages among LPS-induced inflammatory cytokines, by approximately 20% and 45%, respectively.

Conclusion

Our study provides valuable insights into using curcumin and ginsenoside as potential compounds for the effective treatment of radiation-induced injuries and highlights the promising therapeutic benefits of combining these two compounds.
P030 Gut-host metabolites and microbiota interactions in a murine model of checkpoint inhibitor induced colitis
Authors: Liu Z , Alexander J , Zhang R , Lo J , Powell N
Published: January 2024

Background

The advent of Immune Checkpoint Inhibitors (CPIs) has revolutionised cancer treatment by bolstering anti-tumour immune responses. However, the clinical success of CPIs is hampered by immune-related adverse events (irAEs), with colitis posing a common and formidable challenge. Despite this, the precise mechanisms triggering CPI-induced colitis remain elusive. Recognising the influence of gut microbiota on CPI therapy response and colitis development, this study seeks to unravel the intricate details of these mechanisms to facilitate the development of targeted therapies.

Methods

Utilising a murine model of CPI-induced colitis, we combined CPI therapy with faecal microbial transplant (FMT) of colitogenic bacteria. FMT induced colitogenic bacteria in wild-type mice, followed by weekly injections of anti-CTLA4 and anti-PD1 for 3 weeks. Metataxonomic data, obtained through 16S rRNA gene amplicon sequencing, and metabolomic profiles, captured by 1H-NMR spectroscopy, were analysed. Alpha and beta diversity, as well as differential expression of taxa, were assessed using 16S data. Metabolomics data underwent multivariate and univariate analyses to identify metabolic profiles and differential metabolites. Pathway analysis was conducted with MetaboAnalyst. Correlation analyses were performed between bacterial taxa at the family level and metabolite concentrations.

Results

After one week of FMT and CPI treatment, distinct profiles in faecal metabolome and microbiome were observed through Principal Component Analysis (PCA) and Principal Coordinates Analysis (PCoA) for beta-diversity, respectively. Altered metabolites included increased trimethylamine, methylamine, pyruvate, and cytidine, along with reduced tryptophan and proline. Changes in bacterial composition were noted, with increased Bacteroides, Faecalibaculum, Mucispirillum, and Roseburia, and decreased Muribaculum. Metabolic differences persisted after 3 weeks of treatment, revealing additionally decreased acetate, propionate and tyramine. Pathway analysis highlighted profound alterations in tyrosine, alanine, aspartate and glutamate metabolism, pyrimidine metabolism and glycolysis. Desulfovibrionaceae was positively correlated with dimethylamine, glycine and valine. Bacteroidaceae was positively correlated with propionate and butyrate.

Conclusion

This study provides novel mechanistic insights into the metabolite and microbiota-metabolism pathways associated with CPI-induced colitis. The alterations in short-chain fatty acids (SCFA), amino acids, and the SCFA-producing bacteria and colitogenic bacteria shed light on the metabolic changes underlying CPI colitis. Targeting these microbiomes and metabolites could offer promising avenues for enhancing clinical outcomes during CPI therapy.
P031 Microbiota-driven appendiceal B lymphocytes aggravate DSS-induced murine colitis through facilitating colonic CD4+ T cells polarization
Authors: Zhang Y , Xu J , Liu Y , Ke Z , Fang X , Zhang Y , Liu Y
Published: January 2024

Background

The appendix is an immunological organ sustaining a microbiome inoculum that plays a vital role in intestinal mucosal immunity and colonic microbiome stability. Previous studies have proposed the appendix exerts effects on ulcerative colitis (UC)’s pathogenesis, but the nature and basis underlying this connection remain unclear. B lymphocytes account for the largest proportion of appendiceal immunocytes and activate in the experimental colitis model, we systematically investigated how appendiceal B lymphocytes (appB) participate in UC’s pathogenesis and provided rational evidence for appendectomy and B cell immunotherapy being used in UC treatments.

Methods

The chronic colitis model was established in C57BL/6J mice by three cycles of 2% dextran sodium sulfate (DSS) treatment. Bulk RNA sequencing and 16S rRNA sequencing were used to explore appB’s characteristics and the potential causes of appB activation in DSS-induced colitis. Adoptive appB transfer and intravital imaging were used to evaluate appB’s homing and functional features in DSS-induced chronic colitis. Magnetic microbead-based cell sorting and cells experiments in vitro were used to investigate the underlying mechanisms of appB’s activation and effects on DSS-induced chronic colitis. Flow cytometry was used to assess the colonic mucosal adaptive immunity alterations in colitis mice adopted appB.

Results

Firmicutes and Bacteroidetes are the dominant phylum in the normal appendix, while Firmicutes, Verrucomicrobia, and Proteobacteria are prominent in the colitis appendix. The appendiceal IgG+ B cells expended in colitis mice compared with the normal mice; and the DSS colitis appB (DSS_appB), compared with the normal control appB (NC_appB), had much more potential to migrate to the colon and fired the gut flare, accompanied by the increment of CD19+ IgG+ cells, CD4+ IFN-γ+ cells and CD4+ IL-17+ cells in the colonic lamina propria in colitis mice. In vitro, DSS_appB secreted more inflammatory cytokines (IFN-γ, IL-6) than NC_appB under the stimulation of Toll-like receptor agonist analogs (Pam3CSK4, LPS, and ODN1826); DSS_appB facilitated colonic CD4+ T cells to secrete IFN-γ and IL-17 by up-regulating surface marker (CD80/86) and secreting IFN-γ and IL-6.

Conclusion

In DSS-induced chronic colitis mice, appB activated, driven by appendiceal microbiota,migrated to the colonic lamina propria and aggravated colitis through facilitating CD4+ Tcells polarize to Th1 and Th17 by up-regulating CD80/86 and secreting IFN-γ and IL-6 (Figure 1). Our research provides evidence for the connection between the appendix and UC, and we propose that appendectomy or B cell immunotherapy is promising to treat UC.
P034 Peripheral immune cell activity and biopsy gene expression in chronic pouchitis patients treated with tofacitinib: early induction results from the STOPit trial
Authors: Davies J , Amiss A , Khoo E , An Y K , Begun J
Published: January 2024

Background

The Study of TOfacitinib for the treatment of chronic PouchITis (STOPit) is an investigator-initiated, open-label induction, double-blind, placebo-controlled, maintenance withdrawal study to determine the effectiveness of tofacitinib at inducing and maintaining remission in patients with chronic pouchitis. In this interim assessment, we evaluated blood samples and biopsy gene expression collected during the open-label induction period to discover biomarkers of initial clinical response.

Methods

We collected blood (n = 10 pairs, n = 5 non-responders) and biopsies (n = 21 pairs, n = 9 non-responders) at baseline and after 8 weeks of tofacitinib induction therapy (all participants received 10 mg twice daily induction). We classified patients as responders if they achieved a decrease of ≥ 2 in their modified pouchitis disease activity index after 8 weeks. Bulk peripheral blood mononuclear cells (PBMCs) and memory CD4+ T cells (TMEMs) were isolated from whole blood samples. Cytokines induced by stimulation in the presence or absence of tofacitinib were assessed in culture supernatants by bead array (PBMCs stimulated with PMA/Io or LPS, TMEMs stimulated with CD3/CD28 tetramer). Gene expression was measured in biopsies by qPCR in the pouch and pre-pouch ileum at both time points.

Results

At baseline, non-responders produced more IL-17 from both their PBMCs stimulated with PMA/Io and TMEMs stimulated with tetramer than responders, but responders secreted more IL-12p70, MCP, IL-10 and IL-8. Tofacitinib inhibited stimulation-induced IL-10 and IL-6 secretion in PBMCs in both responders and non-responders but failed to reduce IFNγ and TNF secretion selectively in non-responders only. All cytokine responses investigated in TMEMs, including IFNγ and TNF, were inhibited by tofacitinib in both responders and non-responders, suggesting the PBMC cell type refractory to inhibition with tofacitinib in non-responders is not TMEMs. Baseline biopsy ileal gene expression of IL18, HES1 and CXCL10 was decreased in responders compared to non-responders, while MMP7 was increased. Clinical response to tofacitinib led to a reduction in pouch expression of GLIS3, G0S2 and WNT5a from baseline to week 8 whereas these genes were unchanged or increased in non-responders.

Conclusion

Baseline functional immune cell responses and biopsy gene expression may discriminate chronic pouchitis patients' response to tofacitinib therapy and point to a mechanism underlying non-responsiveness. Recruitment for this trial is ongoing, allowing us to bolster and expand our findings with increasing sample sizes.
P050 Paneth cell retinoid X receptor alpha drives metabolic gut inflammation
Authors: Meyer M , Grabherr F , Plattner C , Rieder D , Marteau V , Schwärzler J , Mayr L , Enrich B , Hilbe R , Jukic A , Grander C , Zollner A , Trajanoski Z , Tilg H , Adolph T E
Published: January 2024

Background

The rising incidence of inflammatory bowel diseases throughout the western world could be partially explained by changes in dietary behaviour such as an increased consumption of polyunsaturated fatty acids (PUFA). In previous projects we demonstrated that PUFA induce a Crohn’s disease (CD) like gut inflammation in mice lacking antioxidant GPX4 in their intestinal epithelial cells. Cellular stress within Paneth cells has been shown to drive enteritis in mice. In this project we investigated the role of the PUFA receptor and transcription factor RXRα as driver of PUFA induced metabolic gut inflammation.

Methods

We generated mice lacking GPX4 (GPX4ΔPC) and GPX4 as well as RXRα (GPX4/RXRαΔPC) specifically in Paneth cells and fed them and littermate controls a Western-style diet enriched with PUFA (PUFA-WD). Mice were treated with 9-cis retinoic acid, HX531 or a neutralizing CXCL1 antibody. MODE-K cells (murine IECs) were stimulated with PUFA and were analysed by biochemical methods including ELISA, qPCR, ChIP and RNA sequencing for mechanistical workup in vitro.

Results

When stimulated with PUFA, GPX4 deficient IECs showed an increased activation of RXRα as well as an increased transcription and production of neutrophil attracting chemokine CXCL1, the murine homologue of IL-8. This could be attenuated by ablation of RXRα or treatment with HX531, an RXRα antagonist. By using ChIP we can demonstrate that RXRα is directly binding to the promoter region of CXCL1 upon PUFA stimulation, thereby regulating its transcription. GPX4ΔPC mice develop a CD like enteritis with mucosal to submucosal infiltration of neutrophils, macrophages, B and T cells. GPX4/RXRαΔPC mice were protected from PUFA induced enteritis and gut inflammation could be significantly ameliorated by treatment of GPX4ΔPC mice with RXRα antagonist HX531 or a neutralizing CXCL1 antibody after a PUFA-WD challenge. Moreover 9-cis retinoic acid treatment dose dependently blunted CXCL1 response and attenuated PUFA induced intestinal inflammation in GPX4ΔPC mice, thus indicating a competitive binding of PUFA to RXRα.

Conclusion

Our findings reveal, that PUFA binding to RXRα within Paneth cells regulates the transcription of cytokines and is crucially involved in the development of diet induced metabolic gut inflammation. Future studies are now warranted to prove if RXRα could be a promising new druggable target in human CD patients.
P054 R. intestinalis improves inflammatory bowel disease by preventing gut uric acid absorption through inhibiting METTL3 mediated m6A modification of SLC2A9
Authors: Nie K
Published: January 2024

Background

Inflammatory bowel disease (IBD) is a chronic inflammatory disease with persistent, recurrent, unsatisfactory response, and a poor prognosis. The incidence of Chinese IBD has increased rapidly, and Westernized diets such as purine-rich foods play an important role in its rapid growth. The alteration of purine-urate metabolism by microbiota may contribute to the development of IBD.

Methods

In this study, metagenomic and metabolomic analyses were performed on fecal samples from 30 untreated IBD patients and 30 healthy controls. R. intestinalis was used to observe the effect on serum uric acid levels in mice, respectively. Body weight, colon length, disease activity score, serum uric acid levels, and electron microscopy were all assessed. The RNAseq analysis of intestinal tissue from DSS-induced colitis mice intervened with R. intestinalis and co-cultured with R. intestinalis and the normal intestinal epithelial cell line NCM460, combined with m6A antibody dot blot assay, RM2Target database, and MeRIP sequencing data analysis, provided evidence that SLC2A9 was modified by METTL3, and SRAMP database predicted the potential for SLC2A9 mRNA to be modified by m6A after R. intestinalis intervention.

Results

Multi-omics analysis revealed that the abundance of R. intestinalis was significantly decreased in untreated IBD patients, and metabolomics revealed a correlation between high uric acid levels and disease severity, while the multi-omics integrated analysis suggested a significant negative correlation between R. intestinalis and uric acid. The research team confirmed that R. intestinalis can lower blood uric acid levels in mice and found that R. intestinalis intervention can alleviate the intestinal mucosal inflammation exacerbated by uric acid treatment in a DSS-induced mouse colitis model. Butyric acid has been reported to regulate m6A, and dot blot assays showed that R. intestinalis can reduce the m6A modification level of intestinal epithelial cells, METTL3, and SLC2A9. RM2Target database identified the protein-RNA interaction of the m6A regulator (WER) for SLC2A9 mRNA, and MeRIP sequencing data further indicated that METTL3 can bind to SLC2A9 mRNA through m6A modification. Meanwhile, based on sequence prediction, the research team identified 13 potential m6A modification sites on SLC2A9 mRNA, of which 4 sites had extremely high credibility . The highest credibility site had a score of 0.694, with a modification site sequence of GGACU located on an adenosine nucleotide.

Conclusion

R. intestinalis inhibit the METTL3 mediated m6A modification of intestinal epithelial SLC2A9, reducing the expression of SLC2A9, preventing the epithelial absorption of uric acid, and relieving IBD-related colitis
P060 Increased metabolic activity of faecal Sutterella Wadsworthensis at IBD onset is associated with an increased risk of escalation to biologic therapies - Results from the Birimingham Inception Cohort Study
Authors: Rimmer P , Cheesbrough J , Horniblow R , Tull S , Sharma N , Cooney R , Beggs A , Regan-Komito D , Iqbal A , Iqbal T , Quraishi M N
Published: January 2024

Background

Gut microbiome disruption is an established characteristic of IBD. The lack of treatment naïve longitudinal microbiome data makes it challenging to separate the influence of potential confounders. We present detailed stool metagenomics analysis from a treatment naïve cohort of IBD patients seen in our rapid-access IBD inception clinic.

Methods

Stool collected in DNA Genotek OM-200 kits was collected pre-diagnosis, alongside a faecal calprotectin (FCP). Microbial DNA was extracted. Libraries were prepared using NEBNext Ultra II FS DNA Library Prep Kit, NEBNext Multiplex Oligos and sequenced using an Illumina NextSeq550. Data processing was undertaken using the HUMAnN 3.0 workflow. After quality control, a median of 3.6 million reads per sample were obtained. Abundance was mapped to the ChocoPhlAn database and functional pathways derived from MetaCyc. Diversity analysis was undertaken in Vegan, with MaAsLin2 for taxa differences. A False discovery Rate (FDR) <0.2 was considered relevant, <0.05 highly significant.

Results

A total of 45 CD and 33 UC inception patients were included. There was no significant difference in alpha diversity (Shannon; UC median 2.58[IQR 0.8], CD 2.69[1.04] p=0.36) or differential clustering for beta diversity. Shannon alpha diversity significantly associated with FCP across IBD (n=74, Spearman’s Rho [SR] -0.33 p=0.004) and endoscopic severity in both IBD subtypes (UCEIS n=30 SR -0.37 p=0.047, SESCD n=44 SR -0.35 p=0.016). Across IBD, patients going on to require biologic therapy had a significantly lower baseline Shannon (BioYes 2.26[0.98], BioNo 2.77[0.88] p=0.02, split by IBD subtype in Figure 1).

Across IBD, increased FCP associated with depletion of two strains of Faecalibacterium Prausnitzii (SGB15318, log2 fold change[L2FC] -1.2 FDR 0.09; SGB15342 L2FC -1.1 FDR 0.15) alongside Roseburia Intestinalis (SGB4951 L2FC -0.7 FDR 0.09). R.Intestinalis also negatively correlated with Harvey Bradshaw Index (L2FC -0.93 FDR 0.02), whilst F.Prausnitzii (SGB15342) did with UCEIS (L2FC -2.3 FDR 0.05).

82 metabolic pathways correlated with UCEIS (FDR <0.2, 8 <0.05), with 89 for SESCD (6 <0.05). Increased activity of Sutterella Wadsworthensis associated positively with UCEIS (5 pathways). On initial comparison across IBD, no significant signal was seen for future biologic utilisation. However, if analyses were corrected for baseline inflammation (derived by FCP), increased activity of S.Wadsworthensis was again seen (Table 1).

Conclusion

Reduced alpha diversity at IBD onset associated with increased disease severity and future utilisation of biologic treatment. Increased activity of S.Wadsworthensis associated with an increased risk of future biologic requirement above that explained by mucosal inflammation alone.
P065 The evolution of inflammatory bowel disease can be monitored through multi-omics characterization of gut tissues
Authors: Tout I , Toufiq M , Missous G N , Chin-Smith E , Mathew R , Tomei S , Ibrahim N , Abunahia N S , Mifsud W , Charles A , Ammar A , Al Kaabi S , Van Panhuys N , Al-Mohannadi M J , Mamoun E , Maccalli C
Published: January 2024

Background

The aim of the study is to dissect the mechanisms associated with the evolution of Inflammatory Bowel Disease (IBD), including Crohn’s Disease (CD) and Ulcerative Colitis (UC) in both pediatric and adult patients, along with the severity of the disease and the risk to develop malignancy.

Methods

A multiomics approach, including miRNA (N=700), gene expression (N=800 genes) (nCounter platform; Nanostring) and methylation (Infinium Methylation Epic Bead Chip kit, Illumina), was utilized to characterize the gut tissue biopsies from pediatric (N=20) and adult (N=18) IBD patients. Moreover, a cohort of patients with gut malignancies with history of IBD were also enrolled in the study. Additionally, the role of Natural Killer Group 2D ligands (NKG2DLs) along with different stages of IBD (remission, mild, moderate and severe) were investigated.

Results

Differential miRNA and gene signatures were found by the comparison of tissues from pediatric and adult patients with different stages of the disease. N=83 miRs were significantly differentially detected between pediatric and adult patients (P<0.05). miR-630, miR-493-3p and miR-370-3p were found over-expressed in adult vs. pediatric gut tissues. These miRs are involved in epithelial-to-mesenchymal transition (EMT) through the WNT/β-catenin pathway. These miRNAs can also regulate cell proliferation, cell adhesion and migration and cancer development. Interestingly, miR-20a and miR-34a, which are known modulators of NKG2DLs expression were upregulated in tissues from severe vs. mild disease. The gene expression analysis highlighted, among multiple differentially detected pathways in tissues with different grades of IBD, genes, such as SMAD4, EGR1, CHUCK and JAG1, associated with inflammation, stemness, immune regulation, cell proliferation and migration. Through qPCR analyses, the higher expression of the NKG2DLs MICB was detected in tissues from left or right sides, respectively for both pediatric and adult patients while similar observation occurred for MICA only in adult patients. Soluble NKG2DLs were also detected in the plasma of IBD patients, with in some cases differences (P<0.05) in patients vs. healthy donors. sULBP3 correlated with the presence of active inflammation in adult IBD tissues. Confocal imaging of IBD tissues showed upregulation of stemness-associated markers (e.g., LGR5) and NKG2DLs (ULBP-1 and ULBP-3) in adult vs. pediatric patients.

Conclusion

The ongoing integration of data from multi-omics and the expression of immune-related molecules can lead to the identification of candidate molecular mechanisms associated with the evolution of IBD.
P069 Ultra-processed food intake and dietary insufficiencies are common in adults with Crohn’s Disease in remission. The INTICO 2 Study
Authors: Westoby C , Katarachia V , McDonnell M , Sartain S , Cummings F , Wootton S
Published: January 2024

Background

Adults with Crohn’s Disease (CD) may have dietary restrictions and micronutrient inadequacies. Consumption of ultra processed foods (UPF) has been associated with an increased need for IBD-related surgery and is a growing area of research. This study aimed to investigate the habitual diet of adults with CD during disease remission to understand the nutritional adequacy, diet diversity and diet quality.

Methods

196 adult CD outpatients in remission (Harvey Bradshaw Index <5 and calprotectin <250µg) were recruited from a single UK IBD centre outpatients as part of the INTICO 2 observational study. Seven-day food diaries were electronically recorded using Nutritics® (Libro) software. UPF intake was determined using NOVA Score . Data cleaning was undertaken by a dietitian prior to analysis to identify input errors. NOVA Score1 criteria was utilised to determine UPF intake. Product ingredient lists were examined using supermarket websites and ‘Open food Facts’ by two research dietitians to assign a NOVA score. Consensus on NOVA Scoring was achieved, and a data dictionary was prospectively updated to justify the assigned score.

Results

97% (189/195) of those who underwent food diary analysis had at least one micronutrient below the LNRI. The intake was below the LNRI for a mean of 6 (SD 4.2) micronutrients per subject. The number of subjects who failed to meet the LNRI from diet (with and without inclusion of usual supplements) is listed for each micronutrient in table 1. Of note, 93% (181/195) of subjects failed to meet the SACN recommended 30g of fibre per day (median intake 17.7g). 33% of dietary intake was classified as UPF (Nova 4) and 13% processed (NOVA 3).

Conclusion

Both micronutrient insufficiencies and UPF intake are common in a CD remission population and should be considered alongside one another in studies of dietary intake. UPF intake should be interpreted with caution as NOVA does not account for nutritional content. Blanket recommendations to avoid UPFs need to be carefully considered within this already nutritionally vulnerable population as avoidance of UPF products such as fortified cereals or oral nutritional supplements could lead to increased dietary inadequacies.

References:

Monteiro, C.A. et al, P. 2019. Ultra-processed foods, diet quality, and health using the NOVA classification system. Rome, FAO.
P078 Inflammatory transcriptomic signatures and cell type compositions in colonic mucosa of ulcerative colitis
Authors: Song E M , Jung S A , Joo Y H , Moon C M , Kim J
Published: January 2024

Background

Previous studies have explored transcriptomic profiles associated with active inflammation in patients with IBD patients, but consistent characteristics of gene expression and enriched pathways, particularly in Asian patients with UC, remain unconfirmed. Here, we aimed to assess distinctive transcriptomic changes associated with persistent active mucosal inflammation in Asian patients with UC despite of medical treatment.

Methods

We obtained colonic mucosa biopsies of inflamed and noninflamed tissue from 15 patients with UC and 15 healthy controls and performed RNA sequencing analysis. Utilizing publicly available single cell RNA sequencing (scRNAseq) data, we also identified cell types expressing differentially expressed genes from RNAseq data and performed data deconvolution using CIBERSORTx.

Results

Transcriptomics profiling revealed that the inflammatory transcriptomic signature was highly enriched in the colonic mucosa of UC-active compared to UC-inactive and controls. By incorporating scRNAseq data, the genes upregulated in UC-active were highly expressed in M cells, inflammatory monocytes, and inflammatory fibroblasts, while downregulated genes were prominent in BEST4+ enterocytes and WNT5B+ fibroblasts. Notably, a sub-cluster of enterocytes associated with SAA1, SLC6A14, and DUOXA2 genes exhibited high expression in UC-active. Deconvolution analysis using CIBERSORTx identified significant enrichment of natural killer cells, inflammatory monocytes, Tuft cells, inflammatory fibroblast, WNT2B+Fos-lo1, and pericytes in UC-active.

Conclusion

Our RNAseq analysis not only identified significant gene expression changes but also revealed shifts in cell type proportions associated with persistent inflammation. These findings offer valuable insights for developing novel treatment strategies for UC.
P081 The integration of transcriptomic and microbiomic data links several intestinal bacterial families with key biological functions in the development of postoperative recurrence in Crohn's disease
Authors: Cabrer M , Suau R , Lopez-Siles M , Martinez-Medina M , Aterido A , Julià T , Zabana Y , Calafat M , Clua L , Lorén V , Domènech E , Mañosa M , Manyé J
Published: January 2024

Background

Crohn's disease (CD) is characterized by an imbalance between the microbiota and the host. The severity of CD leads to intestinal resection. However, more than half of the patients not receiving prophylaxis experience post-operative recurrence (POR) within one year. The main objective of this study has been to improve the knowledge of the pathophysiological bases of POR through the integration of transcriptome and microbiome data.

Methods

Samples of inflamed and microscopically non-inflamed ileal mucosa were collected from patients with CD undergoing intestinal resection. The severity of POR was classified according to the Rutgeerts endoscopic index: without recurrence (i0+i1, n=12) and with recurrence (i2+i3+i4, n=8). Samples of healthy ileal tissue were also obtained from patients with colorectal cancer (n=10). Gene expression was evaluated by microarrays and organized into co-expression modules. By sequencing the V3-V4 region of the 16S rRNA gene, intestinal bacteria were identified and classified at the genus and family taxonomic level. The differential expression and correlations between the two sources of omics information were studied. Next, the analysis of pathway enrichment and clustering was carried out according to the severity of the POR and the intestinal area affected or not (graphical abstract A).

Results

The results of the integrative analysis in the uninflamed area of patients without recurrence showed that Actinomycetales negatively correlated with COX6A1 and MTRF1L, related to mitochondrial respiration (GO:0098803; q=0.004) and protein synthesis (R-HSA-72766; q=0.034) (B). These genes and bacteria also played a key role in modules 3 and 25 related to the respiratory chain. In contrast, the inflamed area of recurrent patients showed a positive correlation of the Xanthomonadaceae family in the regulation of neutrophil granules (GO:0035579; q=0.011), with the key participation of the ITGAM transcript (C). This transcript was also related to integrins (R-HSA-216083; q=0.007) in the transcriptomic analyses. In addition, Porphyromonadaceae correlated with LSR and TFF1, involved in epithelium maintenance (GO:0010669; q=0.012) and A group in the Lachnospiradaceae family correlated with CDC20 and CDT1, linked to the cell cycle (q=0.019).

Conclusion

Bacterial families, involved in butyrate production, such as Porphyromonadaceae and Lachnospiradaceae, could influence functions that remember to epithelial regeneration in recurrent patients, while Actinomycetales are associated with a normalization in energy production in uninflamed areas of resections of patients without POR. These findings reveal a potential interaction between the microbiota and the pathophysiology of POR in patients with CD.
P085 Tc17 cells with a distinct immune signature do not correlate with response to therapy with ustekinumab or TNF antibodies in active Crohn’s Disease
Authors: Szentandrasi J , Hockenjos B , Reinscheid M , Thimme R , Bengsch B , Hasselblatt P , Mayer L S
Published: January 2024

Background

IL-17 producing CD8+ T cells (Tc17) expressing a specific immune signature (CD6high, CD39, CD69, PD-1, CD27low) were shown to be associated with active Crohn’s Disease (CD) and inform flare free survival. Here, we addressed the question whether Tc17 cells and their signature also correlate with response of CD patients to therapies with antibodies targeting TNFα or IL-12/23.

Methods

Peripheral blood mononuclear cells were longitudinally collected from CD patients prior to and up to 35 weeks after initiation of therapy with either TNF antibodies or ustekinumab and CD8+ T cells were analyzed by flow cytometry. Results were correlated with clinical outcomes.

Results

We analyzed dynamic changes during treatment in blood samples of 36 CD patients initiating anti-TNF (n = 14) or ustekinumab therapy (N = 22). Baseline Tc17 frequencies differed between ustekinumab and anti-TNF treatment groups, with significantly higher frequencies in the ustekinumab group. This may be due to differences in the baseline patient characteristics; while ustekinumab treated patients were all previously treated with anti-TNF-antibodies, most of the anti-TNF treated patients were naïve to biologics. Indeed, Tc17 frequencies increased during anti-TNF treatment, while Tc17 frequencies were reduced at follow up in the ustekinumab group. However, Tc17 frequencies in the peripheral blood at baseline did not correlate with response to either therapy, nor did the frequencies of Tc17 cells expressing Tc17 signature proteins. The expression of Tc17 signature proteins by IL-17A producing CD8+ T cells did not change significantly during follow up, suggesting that the Tc17 signature was preserved during anti-TNF and ustekinumab therapy. We did not observe major changes in the production of pro-inflammatory cytokines by Tc17 cells during therapy.

Conclusion

Our data suggest that despite playing a pathogenic role in CD, Tc17 cells with a distinct immune signature do not directly correlate with response to therapy with anti-TNF agents or ustekinumab. However, anti-TNF therapy may skew CD8 T cell differentiation towards a Tc17 phenotype, which may have an impact on long term disease outcomes and should be investigated further.
P086 Identification of a Novel Immunometabolic Target and Agonist for PLXDC2 for Amelioration of DSS Colitis Model in Mice
Authors: Abdurahiman S , Mosig R , Cataldi F , Vermeire S , Verstockt B
Published: January 2024

Background

PLXDC2 is expressed on the cell surface of macrophage, dendritic and specific mesenchymal and epithelial cells whose activation shifts cellular metabolism and reduces oxidative stress to rebalance the immune response and decrease inflammation. It’s role in IBD pathogenesis is unknown. PLXDC2 activation improves disease severity in rheumatoid arthritis models1, while loss of PLXDC2 exacerbates the severity of disease in DSS colitis2. PX-04 is a novel, first-in-class, orally active PLXDC2 agonist. Here, we report on the consequences of activating PLXDC2 using PX-04 in an acute DSS colitis model.

Methods

Mice were given DSS in drinking water for seven days to induce disruption of the epithelial layer. At project initiation, mice were 8 weeks of age and began once daily dosing with 20mg/kg of PX-04 24 hours after being placed on DSS. Mice were weighed and scored daily for 15 symptoms of disease (weight loss, diarrhea, rectal bleeding, rectal inflammation, pain, & overall behavior). Colons were collected and digested, and the resultant cellular suspensions were filtered to produce single cell suspensions. Colonic Lamina Propria Immune cells were isolated by Percoll gradient centrifugation. Cells were labeled with mixtures of extracellular (CD45, CD3, CD4, CD8, CD19, NK1.1, CD25, F4/80, CD11b, Gr1, CX3CR1, CD64) and intracellular (Tbet, RORγT, FOXP3, IFNγ, IL17, IL10, TNFα) antibodies in a sequential live staining in 96-well plates. Data was acquired using a FACS Celesta flow cytometer with FACSDiva software.

Results

Oral PX-04 treatment decreased the cumulative disease activity of mice challenged with DSS (FIG 1A). Immunologically, PX-04 affected both the adaptive and innate immune responses. First, PX-04 greatly decreased Th17 cells in the colon, while providing a slight increase to regulatory CD4+ T cells (FIG. 1C). A lower proportion of Natural Killer (NK) cells (FIG. 7D) produced IFNγ. Meanwhile, the proportion of TNF producing dendritic cells was decreased by PX-04 treatment.

Conclusion

Conclusion: PLXDC2 is expressed in cells relevant to IBD pathogenesis. PX-04, which binds to and activates PLXDC2, effectively decreases levels of effector T cells and myeloid cells, as well as overall disease severity in acute DSS colitis, warranting further investigation.

1Tubau-Juni et al. J Immunol 206(Supp)

2Tubau-Juni et al. Sci Rep 10(1)
P102 Neuron-specific gene-2 aggravates colitis and colon tumorigenesis by controlling microbial composition
Authors: Han Y M , Jo K , Lee C H , Yu H , Kim K W , Kang H W , Kim J W , Koh S J
Published: January 2024

Background

Previous studies have shown that Neuron Specific Gene-2 (NSG-2) plays a role in exacerbating murine colitis and colitis-associated tumorigenesis. This study aimed to elucidate the role of Nsg-2 in the pathogenesis of intestinal inflammation by controlling microbial composition.

Methods

The effect of NSG-2 in acute murine colitis was evaluated using both wild-type (WT) and Nsg-2 knock-out (KO) mice. The effect of NSG-2 on colitis-associated tumorigenesis was evaluated using an azoxymethane (AOM) and 3% dextran sodium sulfate (DSS)-induced murine model. The co-housing method was used to assess the role of microbiome in colitis and colitis-associated tumorigenesis. Microbial composition was analyzed using 16S rRNA between the two groups before and after co-housing.

Results

DSS-treated WT mice showed severe colitis compared to DSS-treated Nsg-2 -/- mice. In addition, WT mice exposed to AOM/DSS exhibited higher tumor burdens than Nsg-2-/- mice. After co-housing using WT and Nsg-2 -/- mice, the severity of colitis and colitis-associated tumorigenesis was not different between the two groups, suggesting that microbiome may play a key role in Nsg-2-related aggravation of colitis and tumorigenesis. The microbial taxonomy differed between the two groups in 16S rRNA analysis of stools collected before and after co-housing. Before co-housing, Nsg-2-/- mice showed higher α-diversity than WT mice. Significant differences exist in the Sangeribacter Muris and Lactobacillus Taiwanensis proportion between the two groups. PCoA of beta diversity showed that the microbiome profiles between the two groups were separated before co-housing. However, after co-housing, the difference between the two groups significantly decreased.

Conclusion

Nsg-2 intensifies colitis resulting in colon tumorigenesis through microbial interaction, which suggests that Nsg-2 may have the potential as a therapeutic target for intestinal inflammation.
P130 High serum lipocalin-2 expression at the diagnosis in children with ulcerative colitis: a pilot study
Authors: D'Arcangelo G , Petrella C , Zucconi F , Fiore M , Distante M , Aloi M
Published: January 2024

Background

The identification of new biomarker is a main challenge in inflammatory boweldisease. Lipocalin-2 (LCN-2) is a pleiotropic mediator of various inflammatory processes. We determined the serum levels of LCN-2 in a cohort of newly diagnosed UC children, and we compared them with healthy controls.

Methods

Prospective cross-sectional observational study conducted at the Pediatric Gastroenterology, hepatology, and nutrition Unit of the Umberto I Hospital in Rome and at the IBBC – Institute of Biochemistry and Cell Biology – CNR in Rome. All children aged 6-18 years of age, newly diagnosed with UC at the Pediatric Gastroenterology and Liver Unit, Sapienza University of Rome, were consecutively enrolled. Blood withdrawal was conducted upon diagnosis. Human serum LCN-2 (Cat. No. DY1757) was measured using a sandwich enzyme-linked-immunosorbent assay (ELISA) kits (R&D Systems, Minneapolis, MN, USA), according to the protocols provided by the manufacturer. Clinical, demographic, biochemical and endoscopic data were recorded.

Results

Thirty-two children with a new diagnosis of UC [11 (34%) female, mean age 12,7 ± 4] and 38 age- and sex-matched healthy control [21 (55%) females, mean age 11.71 ± 4] were consecutively enrolled. Serum LCN-2 levels were significantly higher in cases compared to controls (280 ± 152 ng/mL vs 66 ± 55 ng/mL), p < 0,001]. Among UC children, significantly higher LCN2 levels were measured in pancolitis (363.7± 155.2 ng/mL) compared to proctitis, left-sided and extensive colitis (184.9 ± 74.65 ng/mL; p = 0.0003). A significant inverse correlation was observed between LCN-2 and albumin levels at the diagnosis by the Spearman coefficient correlation (r = −0.455; P = 0.03) and a significant direct correlation was observed between LCN-2 and CRP values at the diagnosis (r = 0.44; p <0.05).

Conclusion

Serum LCN-2 levels are significantly higher in children with UC compared to the healthy control group, with the highest levels observed in children with the most extensive disease. Such results as well as and its correlation with the actual disease monitoring tool, make this protein an interesting candidate biomarker.
P135 Mixed probiotics containing Bifidobacterium longum and its metabolites inhibit the formation of NLRP3 inflammasomes by suppressing GBP5 expression in macrophages in ulcerative colitis
Authors: Zhou H , Wu T , Wang F , Zhang T , Lin J , Shi Y , Liang J
Published: January 2024

Background

Ulcerative Colitis (UC) is a non-specific chronic inflammation of the intestine. As an intestinal disease with a gradually increasing incidence in recent years and increasingly valued by medical staff, UC has a close relationship with intestinal flora. At present, domestic and foreign guidelines point out that probiotics can be used to induce remission in ulcerative colitis, but there are no clear instructions on the selection of probiotics, the use of dosage forms, and the dosage of probiotics, and there is a lack of relevant clinical and basic research. This study aims to explore the distinct therapeutic effects of mixed probiotics and individual strains of probiotics (Bifidobacterium longum), as well as elucidate the underlying mechanisms responsible for these variations.

Methods

A total of 90 patients with ulcerative colitis (UC) were enrolled in our center, who were subsequently divided into three groups: control group, single strain probiotic group (Bifidobacterium longum), and mixed probiotic group. After three months of adjuvant probiotic therapy, serum and fecal samples were collected for comparative analysis of therapeutic effects using ELISA and fecal 16s rDNA and metabolome sequence analysis. Mixed probiotic and Bifidobacterium longum were cultured in BHIs in an anaerobic chamber. The supernatant obtained after bacterial cultivation was subjected to metabolome sequence analysis. Human colonic organoids and Caco-2 cells were co-cultured with mixed probiotic and Bifidobacterium longum subsequent to inflammation induction, and cytotoxicity, permeability, and inflammatory cytokine release were assessed. THP1 cells were also to co-cultured with mixed probiotic and Bifidobacterium longum, and expression of virulence genes was analyzed using RT-qPCR, transcriptome sequencing, and western blot. GBP5 gene manipulated cell and mouse model were used to evaluate inflammation and apoptosis.

Results

The utilization of mixed probiotic therapy in patients with UC demonstrated significantly superior therapeutic efficacy compared to those receiving a single strain probiotic (Bifidobacterium longum). Differential metabolite 8-Deoxylactucinwas identified among various groups through the metabolome sequence analysis of clinical patients’ feces, bacterial culture supernatant, and mouse feces. Through in vitro and in vivo experiments, it has been demonstrated that 8-Deoxylactucin exhibits inhibitory effects on the expression of GBP5 in THP-1 cells and ameliorates inflammation.

Conclusion

Collectively, these findings indicate that mixed probiotics and its metabolites suppress GBP5 expression in macrophages in ulcerative colitis.