bluuebunny/crossref_metadata_2025_split huggingface dataset:
DOP54 Guselkumab and golimumab combination induction therapy in Ulcerative Colitis results in early local tissue healing that is sustained through guselkumab maintenance therapy
Authors: Richards D , Vetter M , Germinaro M , Verstockt B , Atreya R , Panes J , Sands B E , Feagan B , McRae B , Cua D , Branigan P , Freeman T C 
Published: January 2024

Abstract

Background
Combination induction therapy with guselkumab (GUS), an interleukin (IL)-23p19 subunit antagonist, and golimumab (GOL), a tumor necrosis factor (TNFα) antagonist, induced higher rates of clinical remission, endoscopic, and histologic outcomes than either monotherapy at Week (WK)12 in patients (pts) with ulcerative colitis (VEGA NCT03662542). Data through WK38 suggested the continued benefit of combination induction even after a transition to GUS monotherapy at WK12. We explored early molecular changes in colon tissue in a subset of pts at WK4 to define mechanistic contributions of each monotherapy and combination; these parameters were evaluated again at WK38 to assess potential carry-over of efficacy.

Methods
Colon biopsies were obtained at baseline (n=195), WK4 (n=42 substudy), and WK38 (n=172). Transcriptional profiles were generated with RNA sequencing (RNAseq). Gene correlation network analysis was applied in conjunction with publicly available single cell RNAseq data to define biologically relevant bulk and cell type-specific transcriptional modules associated with molecular features of disease. Gene set variation analysis (GSVA) was used to quantitatively assess changes in biologic modules with treatment.

Results
At WK4, combination induction (n=10) showed significant (p&lt;0.05) decreases in molecular features compared to GUS (n=19) and GOL (n=13), including transcriptomic modules representing the IL-23 pathway, interferon response, and inflammatory epithelial and myeloid transcriptional states associated with endoscopic improvement at WK4 (Mayo endoscopy subscore of 0 or 1) (combination 5/11, GOL 1/13; GUS 2/19). Reduction (p&lt;0.05) in inflammatory modules persisted through WK38 with combination vs monotherapy induction. Module changes between treatments at WK4 indicated a stronger correlation between GUS and combination (R=0.8; p=2.2e-16) than GOL and combination (R=0.52; p=4.1e-12), with processes associated with epithelial and stromal biology, and mucosal inflammation. In contrast, the combination effect on specific neutrophil/myeloid biology at WK4 was more similar to GOL than GUS, supporting the early role of GOL in targeting innate inflammation.

Conclusion
Combination induction with GUS and GOL showed significant reductions in major inflammatory features of disease as early as WK4 which persisted through WK38 with GUS maintenance. Correlative analysis supports the role of GUS as the primary driver of tissue healing, with GOL further contributing to innate inflammatory activity, which demonstrate differential and complementary mechanisms of action of TNFα and IL-23p19 subunit blockade.
DOP56 Fibrosis-related transcriptome unveils a distinctive matrix remodelling pattern in penetrating but not in stricturing ileal Crohn's Disease
Authors: Tavares de Sousa H , Ferreira M , Gullo I , Rocha A M , Oliveira C , Carneiro F , Magro F 
Published: January 2024

Abstract

Background
Fibrosis underlies most of CD complications requiring surgery, such as intestinal strictures and penetrating events. We previously demonstrated there were no histopathological differences on transmural fibrosis and fibromuscular changesbetween penetrating and stricturing ileal CD. This study aimed to investigate and compare the fibrosis-related transcriptomic profiles of penetrating and stricturing ileal CD.

Methods
Using Nanostring technology and comparative bioinformatics, we analysed the expression of 787 genes covering the core pathways and processes involved in fibrosis, in 34 ileal surgical specimens, 11 B2 and 23 B3, the latter encompassing 12 with associated stricture(s) (B3s) and 11 without (B3o). Each patients’ tissue was retrieved from three FFPE 20mm-sections. Quality control of extracted RNA was performed according to Nanostring parameters and principal component analysis (PCA) graph for the distribution analysis. For the selection of the differentially expressed genes (DEGs) we adopted a P-adjusted &lt;0.05 and a Fold Change ≤-1.5 or ≥ 1.5. Statistical analysis adopted level of significance at 5%.

Results
We included 34 patients, in a well-balanced sample for age (at diagnosis and at surgery), gender, CD localization, perianal disease and therapy. Inflammation and fibrosis histopathological scoring were similar (median of 3 and 1, respectively), in the three groups. No DEGs differentiated B3s and B3o subgroups, which were grouped as B3. The B2 and B3 groups showed a very good clustering on the gene heatmap, which uncovered 30 DEGs between the 2 groups, all being upregulated in B3 and, conversely, downregulated in B2 patients – Figure. More than half of the upregulated genes have been established or partially involved in CD fibrogenesis (yellow-box and line, respectively), while 8 DEGs have been implicated in fibrosis in other organs (brown-line box) and 3 others having an anti-fibrotic role in other tissues (green-line box). Importantly, the vast majority of the most significantly active biologic processes and pathways in the B3 group were related to response to TGF b and matrix organization, production and degradation, unveiling a continuous process of tissue healing and destruction distinctive of this aggressive phenotype.

Conclusion
Despite the histopathological similarities of fibrosis between stricturing and penetrating ileal CD, their fibrosis-related transcriptomic profiles are distinct. Penetrating CD was remarkable for the activation of all the unveiled DEGs and of biologic pathways concerning permanent matrix production and degradation. Hence, although fibrosis underlies both penetrating and stricturing CD, therapeutic targets may differ even in end-of-stage disease.
P004 Functional interrogation of the stem cell graft reveals heterogeneity that may determine clinical outcomes after hematopoietic stem cell transplant for refractory Crohn's Disease
Authors: Guisado D , Talware S , Singh S , Fozilov E , Colombel J F , Cho J , Cohen L 
Published: January 2024

Abstract

Background
For the treatment of refractory Crohn’s disease (CD) autologous stem cell transplant (auto-SCT) is unparalleled in its ability to induce clinical and endoscopic remission. Auto-SCT is unique as a cellular therapy aimed to reset immune pathophysiology to a pre-disease state using hematopoietic stem cells (HSC). To date no studies in any cohort have defined the mucosal and peripheral immune response to auto-SCT or analyzed HSC functions.

Methods
Patients with CD were enrolled in a Phase IIa study of auto-SCT (NCT03219359) with 16 patients transplanted (2018-2023). Patients underwent a standard mobilization regimen with cyclophosphamide and G-CSF. Mononuclear cells from the mobilized stem cells of 9 patients were isolated and then analyzed for phenotyping using flow cytometry. For xenograft studies, CD34+ selected stem cells from 6 patients were injected into NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice to assess engraftment and differentiation potential by flow cytometry of cells from bone marrow at multiple time points.

Results
At 6 months from auto-SCT, 15/16 patients had an endoscopic response (Total SES-CD decrease ≥ 50%) and 11/16 were in endoscopic remission (total SES-CD ≤ 4, no sub-score &gt;1). Phenotyping of the stem cell graft demonstrated variability between patients but overall there was predominance in short-term HSCs and lineage committed progenitors rather than long-term HSCs (LT-HSC). There was variable engraftment of the CD34+ cells that were transplanted in NSG mice and 2 of the patients who had inadequate engraftment also were noted to have suboptimal clinical results, 1 with disease relapse and 1 without clinical response after auto-SCT. Of those with engraftment, we found a predominance of myeloid populations emerging early and sustaining and emergence of B cells at 8 weeks.

Conclusion
Here we report the first ever study of the stem cell graft using a xenograft model to understand therapeutic mechanisms of auto-SCT for refractory CD. These results show tremendous functional variation in mobilized stem cell populations from patients with CD and a complete lack of LT-HSCs, which have greater self-renewing potential. This suggests that the efficacy of auto-SCT for refractory CD may be due in part to the HSCs transplanted and not just the conditioning regimen (i.e. chemotherapy). These studies suggest the response to stem cell transplant may depend on the presence of functional HSC populations and perhaps the lack of long-term response is due to the lack of LT-HSC in all patients.
P011 Altered Expression and Enzymatic Activity of Xanthine Oxidase in Inflammatory Bowel Disease
Authors: Di Petrillo A , Onali S , Era B , Raho N , Murtas D , Maxia C , Favale A , Eleonora G , Pinto S , Fais A , Fantini M C 
Published: January 2024

Abstract

Background
Xanthine oxidase (XO) is involved in the last steps of purine metabolism; indeed, it catalyzes the oxidation of hypoxanthine to xanthine and xanthine to uric acid (UA) with the production of superoxide anion. The accumulation of UA has been shown to initiate the inflammatory process through NLRP3 inflammasome and the production of reactive oxygen species (ROS) contributes to inflammation-related tissue damage. In murine models of colitis, XO inhibitors, allopurinol or febuxostat, reduced the expression of proinflammatory cytokines supporting the involvement of XO in the intestinal inflammatory process. This study aims to conduct a thorough investigation of the role XO plays in IBD pathogenesis and to explore the potential of XO inhibition as a therapeutic strategy.

Methods
Biopsies from the colon of patients with ulcerative colitis (UC) and Crohn’s disease (CD) were collected at University Hospital of Cagliari. Disease activity was assessed using Mayo and SES-CD scores. XO expression levels were evaluated using Real-Time PCR, validated by GEO database (GSE11223, GSE117993). XO enzymatic activity was quantified spectrophotometrically in protein extracts from the biopsies. XO and NLRP3 inflammasome activation pathway, including NLRP3, ASC, and CASPASE1, was investigated using immunohistochemistry (IHC). In vitro, Caco2, HT29, SW480 cells were treated with xanthine, XO inhibitors (allopurinol, febuxostat) to assess XO inhibition on NLRP3 activation, analyzed by immunocytochemistry (ICC).

Results
Biopsies from the sigmoid colon of 15 HC, 20 UC, and 15 CD patients, as well as from the ileum of 10 HC and 10 CD, were analyzed. Increased XO expression was found in UC and CD sigmoid mucosa (Relative Quantification [RQ]: 1.93 vs. 0.99, p=0.004), and CD ileum (RQ: 2.72 vs. 1.26, p=0.004), confirmed by GEO data. XO activity was higher in UC and CD sigmoid (3.86 nmol UA/min/mg in UC, 4.82 in CD vs. 1.03 in HC, p&lt;0.001) and CD ileum (8.41 vs. 1.83 in HC, p&lt;0.001). IHC confirmed that XO and the NLRP3 inflammasome pathway are more highly expressed in IBD patients than HC, predominantly within epithelial cells. In vitro studies on Caco2, HT29, and SW480 cells demonstrated that suppressing XO activity with XO inhibitors leads to the inhibition of the NLRP3 pathway.

Conclusion
XO expression was higher in IBD pts than in HC individuals. Alongside this increase in XO activity, there was a concurrent hyperactivation of the NLRP3 inflammasome. The inhibition of XO activity in cell lines resulted in reduced activation of the NLRP3 inflammasome pathway. These findings support the hypothesis that XO plays a critical role in the pathogenesis of IBD and highlight its potential as a therapeutic target.
P017 An activity-based chemiluminescence assay targeting granzyme A for the detection of Inflammatory Bowel Diseases
Authors: Scott J , Thompson E , Cheng Z , David M , Gordon D , Medive-Tapia L , Jones G R , Hall R , Chuah S , Drury B , Rossi A , Shabat D , Ho G T , Vendrell M 
Published: January 2024

Abstract

Background
Inflammatory Bowel Diseases (IBD) represent a pressing global health concern lacking precise diagnostic tools for accurate patient stratification. Conventional methods, such as colonoscopy are extremely invasive whilst generic inflammatory markers (such as CRP or Calprotectin), are not specific enough and pose limitations in assessing disease activity, urging the need for novel, non-invasive diagnostic approaches which can rapidly inform clinicians on the inflammatory status of a patients bowels.

Methods
Our study focused on developing a non-invasive chemiluminescence assay targeting active granzyme A (GzmA) in stool supernatants. Employing a multifaceted approach encompassing functional, chemical, and imaging assays on human cells and gut tissue biopsies, we discerned elevated GzmA levels predominantly secreted by CD8+ T cells in inflamed gut tissues.

Results
The design of GzmA-INH, a specific inhibitor for GzmA, underscored the prevalence of active extracellular GzmA and it’s role in driving inflammation in the guts - revealing promising potential as a novel IBD biomarker. Further, we synthesized GzmA-CL, a highly reactive and selective chemiluminescent reporter for GzmA. Integrating GzmA-CL into a robust antibody-coated 96-well plate assay (Figure 1a), we analyzed 30 IBD biosamples along with healthy controls. Notably, substantially higher GzmA levels were observed in IBD patients, with discernible distinctions between ulcerative colitis (UC) and Crohn’s disease (CD) (Figure 1b). These levels correlated positively with established biomarkers, including faecal calprotectin and immune cell counts.

Conclusion
Our investigation highlighted active extracellular GzmA as a noteworthy biomarker in gut inflammation. The rapid, selective, and sensitive nature of our chemiluminescence assay presents promising prospects for precise IBD diagnosis and advancements in personalized medicine.
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

Abstract

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

Abstract

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

Abstract

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.