bluuebunny/crossref_metadata_2025_split huggingface dataset:
DOP88 Lipids drive myofibroblast activation and the epithelial-mesenchymal transition process in Crohn’s Disease
Authors: Rizzo G , Wozny M , Pineda Chavez S E , Mori G , Carriles Linares A , Morosi L , Martano G , Ghisletti S M L , Gandolfi F , Vallelonga V , Loy L , Bezzio C , Spaggiari P , Dal Buono A , Gabbiadini R , Carvello M , Spinelli A , Repici A , Armuzzi A , Vetrano S 
Published: January 2024

Abstract

Background
Fibrostenosis represents one of the major clinical complications of Crohn’s Disease (CD). However, the molecular mechanisms underlying intestinal fibrogenesis remain poorly understood and anti-fibrotic therapy has not yet been developed. Growing evidences supports that oxidative stress and the epithelial-mesenchymal transition (EMT) process contribute to the development and progression of fibrosis. Here, we assessed the role of lipids in fibrogenesis and whether mitochondria dysfunction can support fibrosis formation.

Methods
Spatial metabolomics analysis and adipophilin (ADFP) staining were performed on surgical specimens from fibrotic ileum and the surrounding region (healthy) derived from CD patients. Fibroblasts isolated from fibrotic and healthy ileum and human induced pluripotent stem cell-derived intestinal organoids (iHOs) were treated for 1-3 weeks with lipid mix (2%) and analyzed by RNA sequencing. Pathways significantly modulated were validated by RT-PCR, WB, FACS analysis, and proliferative, migratory and contraction assays, and mitochondrial functions by Seahorse analyzer.

Results
Spatial metabolomics revealed a significant accumulation of lipids characterized by very long-chain fatty acids in the fibrotic area compared to surrounding healthy regions, which was further confirmed by ADFP staining. RNA-seq analysis showed overlapping dysregulated pathways between healthy fibroblasts stimulated with lipids and fibrotic fibroblasts, in particular genes related to cell cycle, cytoskeletal reorganization, migration, contractility and cell metabolism (oxidative stress) which were further confirmed by functional assays. Among the genes related to oxidative stress, lipid stimulation significantly decreased NFE2L2 and promoted an increase in CPT1A, gene encoding for a fatty oxidation mediator. Moreover, lipid-stimulated fibroblasts showed a reduction in mitochondrial respiration, particularly basal and maximal respiration, and spare respiratory capacity by Seahorse analysis. On the other hand, long exposure of iHOs to lipids promoted the EMT process through the upregulation of TWIST, ZEB, VIM, and S100a4.

Conclusion
Lipid stimulation induced myofibroblast activation through oxidative stress and alteration of mitochondrial metabolism, and increased the expression of EMT transcription factors in intestinal epithelial cells (iHOs) that may support fibroblast-mediated pro-fibrotic activity. These data indicate that regulators of fatty acid oxidation involved in the fibrotic process may represent new potential targets for the treatment of fibrostenotic CD patients.
DOP89 Unfolded protein response controls group 3 innate lymphoid cells in IBD
Authors: Cao S , Colonna M 
Published: January 2024

Abstract

Background
Group 3 innate lymphoid cells (ILC3s) have recently emerged as important regulators and potential drug targets for IBD. However, the response of ILC3s to environmental stimuli during intestinal inflammation remains elusive. IRE1a-XBP1 serves as the regulatory hub of endoplasmic reticulum stress and the unfolded protein response (UPR) that plays a vital role in intestinal inflammation.

Methods
We generated conditional knockout Ire1aflo/flox,Rorc-Cre (Ire1aΔRorc) and Rag-/-Ire1aΔRorc mice with Ire1a deleted in ILC3s. Intestinal infections were induced by Citrobacter rodentium and Clostridium difficile. ILC3s-derived cytokines including IL-22 is crucial for host defense in the acute phase of C. rodentium and C. difficile infection. Dextran sodium sulfate (DSS) was given in drinking water to induce acute colitis. We recruited patients with active Crohn’s disease (CD) and collected ileal biopsies during routine colonoscopy before starting ustekinumab, a non-selective anti-IL-23 antibody. Mucosal ILC3s were isolated for analysis of ILC3s by flow cytometry.

Results
In murine intestinal ILC3s, the activity of IRE1a-XBP1 pathway follows a robust 24h circadian rhythm which parallels that of clock genes and cytokines including IL-22. XBP1s is activated in ILC3s ex vivo by vasoactive intestinal peptide, which was previous shown to orchestrate the circadian expression of IL-22 by ILC3s. IRE1a-XBP1 in intestinal ILC3s is further activated in response to IL-23 and colitis in mice, as well as in inflamed IBD tissues compared to normal tissues from healthy individuals. We further showed that IRE1a-XBP1 activation in stimulated ILC3s requires mitochondrial reactive oxygen species (mtROS) (Fig 1). Using the Ire1aΔRorc mice, we demonstrated that IRE1a-XBP1 is critical for cytokine expression by ILC3s. Ire1aΔRorc mice are highly susceptible to acute C. rodentium and C. difficile infection as well as acute DSS colitis (similar vulnerabilities were found in Rag-/-Ire1aΔRorc mice compared to Rag-/-Ire1aflox/flox littermates). Ire1aΔRorc mice exhibit reduced IL-22 and IL-17A in ILC3s and impaired epithelial barrier function with diminished expression of mucins and antimicrobial peptides. The susceptibilities of Ire1aΔRorc and Rag-/-Ire1aΔRorc mice to colitis were rescued by administration of recombinant IL-22 (Fig 2). Moreover, the frequency of XBP1s+ ILC3s in ileal mucosa from CD patients before initiation of ustekinumab positively correlates with response to therapy.

Conclusion
We demonstrate that a non-canonical mtROS-IRE1a-XBP1 pathway augments cytokine production by intestinal ILC3s. We also identify XBP1+ ILC3s as a potential biomarker for predicting responsiveness to anti-IL-23 therapies in IBD.
DOP90 Amphiregulin promotes colitis-associated intestinal fibrosis through activation of PI3K/AKT signaling in Intestinal fibroblasts
Authors: Wang S , Zhang H , Zhao X 
Published: January 2024

Abstract

Background
Intestinal fibrosis is one of the most threatening complications of Crohn’s disease (CD). Amphiregulin (AREG) expression is increased in patients with CD with fibrosis.

Methods
The role of AREG in activating intestinal fibroblasts and driving fibrogenesis is largely unexplored. In this study, fibrotic and nonfibrotic tissues were obtained from CD patients undergoing surgical resection while normal intestinal tissues were obtained from patients with diverticulum of small intestine. Fibrosis index score for fibrosis were assessed by Masson staining. AREG, collagen Ⅰ and α-SMA expression were determined in intestinal tissues of patients with CD with fibrosis or without fibrosis by qRT-PCR. Wild-type (WT) and Areg knockout (Areg-/-) mice were induced into intestinal fibrosis by dextran sulfate sodium (DSS). We induced intestinal fibrosis in WT mice using a chronic DSS-induced colitis model with or without AREG intraperitoneal injection. The fibrotic indicators in colons were evaluated. Fibroblasts were isolated from fibrotic intestinal surgical resections of patients with CD. The effect of AREG on activation and proliferation in human intestinal fibroblasts was determined. RNA-sequencing of human intestinal fibroblasts treated with or without AREG was performed.

Results
AREG expression was increased in intestinal fibrotic sites compared with nonfibrotic sites from patients with CD. Mice with Areg deficiency represented with severer intestinal inflammation but alleviated fibrosis in colons. Although additional given AREG attenuated intestinal inflammation in mice with DSS-induced chronic colitis, it promoted more severe intestinal fibrosis. AREG promoted human intestinal fibroblast activation and proliferation by activating PI3K/AKT signaling. PI3K inhibitor suppressed AREG-induced fibroblast activation and proliferation, thus lightened intestinal fibrosis in mice. Besides, AREG promoted secretion of LPA (Lysophosphatidic acid) from intestinal fibroblast, which activated fibroblast through LPAR3/PI3K/AKT pathway.

Conclusion
These findings reveal that AREG promotes intestinal fibrotic responses in experimental colitis and human patients with CD by promoting fibroblast activation and proliferation through PI3K/AKT pathway. Moreover, LPA secreted from intestinal fibroblast induced by Areg is involved in formation of CD intestinal fibrosis. Thereby, PI3K, AKT, LPA might serve as potential therapeutic targets for fibrosis in CD.
P003 Metabolic Reprogramming in Intestinal Fibrosis: Integrating Metabolomics, Single-Cell Transcriptomics and Isotope Tracing
Authors: Zhang Y , Zhuang H , Gu Y , Zou D 
Published: January 2024

Abstract

Background
This study aims to systematically elucidate the metabolic landscape of intestinal fibrosis in Crohn's disease, revealing the key reprogrammed metabolic pathways in the fibrotic microenvironment.

Methods
We collected normal and fibrosis tissues from 122 Crohn's disease patients undergoing surgery for fibrosis-induced intestinal obstruction, and established DSS/TNBS-induced murine intestinal fibrosis model. To measure metabolites level in normal and fibrotic tissues, untargeted metabolomics was performed. C13-labeled glutamine was used for metabolic tracing to delineate metabolic fluxes in vitro and in vivo. We performed integration analysis by merging metabolomics and metabolic tracing data with established single-cell transcriptomic profiles. Isolation and further validation of human intestinal stromal cells were carried out, and gene expression was assessed using qRT-PCR and immunofluorescence.

Results
Significant metabolic heterogeneity was observed between normal and fibrotic tissues, with differential metabolites enriched in glucose and amino acid metabolism. Notably, intermediates of glycolysis were significantly increased, while tricarboxylic acid (TCA) cycle intermediates were decreased in fibrotic regions (p&lt;0.05). Amino acids were broadly downregulated in fibrosis, whereas glutamate and glutamine were highly enriched (p&lt;0.05). Mechanistic insights into these changes were provided by isotope tracing, revealing enhanced glycolysis and weakened TCA flux attributed to transition from TCA intermediate α-KG to glutamate. Moreover, catabolism of glutamine to glutamate was enhanced. Glutamate from above pathways converted to proline, which is the ingredient of extracellular matrix (ECM).
Single-cell transcriptomic analysis indicated a high enrichment of genes related to glutamine metabolism in stromal cells, with significant upregulation of enzymes such as GLS (Glutaminase) and PYCR1 (Pyrroline-5-carboxylate Reductase 1) involved in glutamine and proline synthesis. Immunofluorescence and qRT-PCR confirmed the upregulation of GLS and PYCR1. Further experiments using primary stromal cells and murine models demonstrated that inhibiting GLS activity reduced ECM synthesis, alleviating intestinal fibrosis.

Conclusion
In intestinal fibrosis, glycolysis and glutamine catabolism enhanced. Stromal cells utilized the reprogrammed metabolic pathway to synthesize glutamate, thereby promoting pathological ECM anabolism. Targeting the key enzyme GLS represent a potential therapeutic strategy for intestinal fibrosis.
P006 Development of a Crohn's disease molecular activity score to identify region-specific chronically inflamed and non-inflamed processes
Authors: Richards D , Venkat S , Terry N A , Vetter M , Gomez M , Freeman T C , McRae B , Feagan B , Reinisch W , Branigan P 
Published: January 2024

Abstract

Background
Crohn’s disease (CD) is characterized by significant heterogeneity in disease location and other clinical factors, making it challenging to define the underlying molecular mechanisms associated with disease. Here, we provide a detailed molecular characterization of baseline ileal and colonic tissue from patients enrolled in the GALAXI Ph2b study of guselkumab in CD (NCT03466411) and explore the association of regional molecular profiles with endoscopic and histologic severity.

Methods
Baseline biopsies from 241 patients were obtained from rectum (R), splenic flexure (SF), and terminal ileum (TI). Paired biopsies within a region were processed for histology and RNA sequencing (RNAseq). Cell-type specific transcriptional modules were used to evaluate differential expression and to create a disease severity axis. Principal component analysis (PCA) was applied to modules with the highest correlation with paired Global Histologic Activity Scores (GHAS) per region and the resulting PC1 was used to define an objective tissue-based molecular activity score (MAS) to identify inflamed and uninflamed tissue. Inflamed tissue was used to assess regional molecular profiles which were correlated with paired regional histology scores defined by GHAS and endoscopic severity defined by Simple Endoscopic Score for CD (SES-CD).

Results
Modules specific to inflammatory myeloid and epithelial transcriptional states, and IL-23 and interferon response pathways, were highly correlated with paired GHAS. Applying this subset of modules as a MAS revealed a bimodal distribution of inflamed and non-inflamed biopsies. MAS identified 114/241 (47.3%), 85/229 (37.1%) and 114/229 (49.8%) inflamed biopsies in the R, SF, and TI, respectively. Regional molecular profiles of inflamed biopsies demonstrated increased mRNA expression of IL-23p19 (IL23A) and Fc-γ receptor 1 (CD64) (p&lt;0.001), and enrichment of inflammatory modules across locations. Local MAS per region showed highest correlations with paired baseline GHAS, followed by SES-CD in the segment (Figure 1 and Table 1). Notably MAS provides higher resolution to identify representative biopsies for affected regions.

Conclusion
Application of the MAS to identify inflamed tissue enabled identification of key immune and inflammatory related processes across the ileum and colon that were associated with the IL-23 pathway, and baseline regional endoscopic and histologic severity. By identifying inflamed baseline samples, molecular changes associated with treatment can be more accurately defined in future analyses to better understand regional heterogeneity in CD.
P013 Magnetic Resonance Imaging-Based Average T2 Signal as A Non-Invasive Measure of Fibrosis In Enteric Crohn’s Disease
Authors: Naim I , Clarke C , Hoad C , Jinnah N , Mukherjee A , Gowland P , Mougin O , Menys A , Bard A , Moran G W 
Published: January 2024

Abstract

Background
In Crohn’s Disease (CD), 10% of patients present with fibrostenosis with a further 10% progressing from inflammatory to a fibrostenotic disease behaviour over 7 years. 40% of fibrostenotic CD patients need surgery at 5 years compared to ~10% with inflammatory disease. Present non-invasive fibrosis biomarkers lack the required accuracy needed to be used routinely for clinical decision making. T1, T2, diffusion weighted imaging, motility and bowel volumes have shown promise in the non-invasive measure of fibrosis.

Methods
The primary objective of this study is to undertake biological validation of MRI measures as independent imaging biomarkers of histological fibrosis with the primary outcome being the correlation between MRI measures and histological fibrosis measures. GI-Seg (Motilent, London, UK) is an image analysis tool used to segment the gastrointestinal tract on T2W MRI scans.
Patients with CD histological diagnosis and in need of surgical resection for stenosis attended a one-hour 3T MRI scanning session within 12 weeks prior to their intestinal resection surgery. Histological samples were scored by a specialized gastrointestinal pathologist for inflammation, fibrosis, and muscular hypertrophy. An expert radiologist with the help of the histopathologist located the strictures on DWI, quantitative T2&T2 scans, and T2 weighted clinical MRI scans. GI-Seg was used to generate a 3D model of the stricture on T2W scans measuring bowel wall volume and average signal intensity of each stricture. Pearson correlation coefficient was used to quantify the correlation between the variables.

Results
A total of 10 CD patients, encompassing 20 strictures, were included in the study. The analysis revealed that the average T2 signal has a significant correlation with global fibrosis scores (r(20) = .563, p = .010) (Figure) and total inflammation scores (r(20) = .451, p = .046). Additionally, stricture volume showed a fair correlation with chronic inflammation component scores (r(20) = .453, p = .045). Notably, quantitative T1 values correlated with total inflammation scores (r(17) = .493, p = .045).

Conclusion
The study reveals fair correlations between the T2 signal average and both global fibrosis and global inflammation scores, as well as between stricture volume and chronic inflammation in CD strictures. While these findings indicate that the GI-seg tool's measurements of bowel wall volume and signal intensity have potential utility as imaging biomarkers for fibrosis and inflammation, the results also underscore the necessity for additional markers that are specifically correlated with fibrosis. Such markers are essential to distinctly identify fibrotic changes, independent of inflammatory processes.
P018 Inequalities in healthcare access, experience and outcomes in adults with Inflammatory Bowel Disease: A scoping review
Authors: Hawkins R , Zia M , Hind D , Lobo A , Ezaydi N 
Published: January 2024

Abstract

Background
Inflammatory Bowel Diseases (IBD) are long-term conditions requiring lifelong access to, and interactions with health services. Accumulating evidence of inequalities in health care access, experience and outcomes for individuals with IBD is apparent. However there is no existing review that systematically maps these inequalities.The purpose of this review was to map and describe the inequalities in healthcare access, experiences, and outcomes of care for adults with IBD, to identify research gaps and future research priorities in this area.

Methods
A scoping review was conducted to retrieve quantitative, qualitative, and mixed methods evidence from three electronic databases (EMBASE, Medline and CINAHL). English written articles published between 2000 and 2023 were systematically searched using a search strategy. The Joanna Briggs Institute protocol for scoping reviews was followed to perform data extraction and analysis.

Results
1343 records were identified. 51 studies met the criteria for inclusion. Studies addressed ethnic disparities of healthcare (33/51), inequalities driven by socioeconomic differences (12/51), rurality (7/51), gender (3/51), age (2/51), culture (2/51), literacy (1/51) and sexuality (1/51). The interrelationship of disparities relating to outcome, access and experience is illustrated in Figure 1. Poorer health outcomes (42/51 studies) were described in those from Black, Asian, and Hispanic ethnic groups in relation to hospital admission, readmission, emergency room attendance, length of stay, IBD-related complications, post-operative complications, treatment delay and length of stay. Deprivation, insurance status in US populations, rural location and age also affected outcome. Access inequalities (24/51 studies) were described to services, therapies and surgery but also to educational opportunities. Poor experience of care (8/51 studies) included appointment issues, lack of cultural understanding and fragmented care. A lack of research was found in the LGBTQIA+ community (1/51). No research was found to investigate inequalities in IBD patients with learning disabilities or autism.

Conclusion
Inequality affecting a wide range of population characteristics affects outcomes, access and experience in IBD care. Services need to be organised and delivered in a way that addresses this. Further research, including qualitative methods, is particularly needed to understand health experiences of underserved patient populations with IBD. The lack of research amongst LGBTQIA+ individuals, and with learning disabilities, poses a risk of creating inequalities within inequalities.
P020 Effects of Anti-TNF Treatment on Browning of Mesenteric Adipose Tissue with Crohn's Disease
Authors: Montfort-Ferré D , Vaño-Segarra I , Boronat-Toscano A , Menacho M , Moliné A , Cepero C , Mañas M J , Caro A , Serena C 
Published: January 2024

Abstract

Background
Crohn's disease (CD) is characterized by the expansion of mesenteric fat, known as creeping fat (CrF), to the inflamed segments of the intestine. Our group observed that CrF showed a conversion to beige tissue, a phenomenon known as "Browning", characterized by an elevated gene expression of thermogenin (UCP-1), the gene characteristic of browning1. A recent study also showed that when browning was induced in mice in a TNBS colitis model, mesenteric hypertrophy improved and inflammation in adipose tissue was reduced2. The aim of this study is to assess whether the administration of anti-TNF biologic treatment (infliximab) in patients with CD is related to the increase in adipose beige tissue in the CrF.

Methods
Immunohistochemistry staining of UCP-1 was conducted on CrF samples from CD patients receiving anti-TNF treatment (≥6 months) and anti-TNF-naïve patients. Additionally, we analysed the gene expression of beige adipose markers in CrF from CD patients treated with anti-TNF drugs, comparing them to those untreated. UCP-1 gene expression was also studied in adipose-stem cells isolated from CrF. Ongoing experiments involve treating adipose stem cells with infliximab (10ug/ml) for 24 hours to assess the potential conversion of white to beige adipose tissue.

Results
The study revealed a significant increase in UCP-1 staining in CrF samples among patients receiving anti-TNF treatment compared to anti-TNF-naïve CD patients (Fig 1.A). This trend was consistently observed in mesenteric CrF tissue. Beige markers TMEM26 and CPTB1 exhibited significant increases in anti-TNF-treated CrF, and other beige tissue markers showed a notable trend (Fig 1.B). Adipose stem cells displayed a significant increase in UCP-1 gene expression in anti-TNF-treated CrF compared to untreated patients (Fig 1.C). Exciting preliminary data suggest that infliximab treatment increases UCP-1 and the anti-inflammatory cytokine IL-4 gene expression in adipose-stem cells from active CD patients (Fig 1.D). This novel data implies a direct relationship between anti-TNF treatment and heightened browning in white adipose tissue of CD patients.

Conclusion
This study reveals a notable increase in UCP-1 gene expression in adipose tissue CrF of CD patients undergoing anti-TNF treatment. The findings suggest that anti-TNF therapy may trigger the transformation of white adipose tissue into the "beige" type, potentially mitigating inflammation. Browning CrF presents a promising therapeutic avenue for CD management, warranting further investigation.
Ref:
1. D. Monfort-Ferré et al. J Crohn Colitis., 2022;16(10):1571–83,
2. Zuo L et al. J Crohn Colitis. 2023 21;17(8):1179-1192.