Introduction

The BIOSTOP trial is a Norwegian, multicenter study with a prospective, randomized, controlled, and open-label design. The trial was initiated to evaluate the two-year outcomes following the withdrawal of tumor necrosis factor-alpha (anti-TNF) antibodies in patients with ulcerative colitis (UC) who are in sustained clinical and endoscopic remission.

Aims & Methods

The main aim was to identify transcriptomic differences in colon biopsies associated with increased risk of relapse with anti-TNF withdrawal. We also wanted to increase our understanding of the underlying pathological mechanisms of relapse after anti-TNF withdrawal and treatment failure during continued treatment.
Between June 2017 and December 2020, 174 ulcerative colitis patients in confirmed clinical and endoscopic remission were enrolled and randomized, with 86 assigned to continue anti-TNF therapy (control group) and 86 to discontinue treatment (intervention group). Mucosal biopsies were collected at baseline, two-year follow-up, and at point of relaps
Bulk mRNA-sequencing was performed on RNA extracted from rectal biopsies. For patients in remission, samples from baseline and two-year follow-up were analyzed. For those who relapsed or experienced treatment failure, samples from baseline and point of relapse were analyzed. Differential gene expression was performed using DESeq2, and pathway enrichment analysis was assessed with clusterProfiler.

Results

There were no significant differences found at baseline between patients that relapsed (n=45) vs those that stayed in remission (n=41) after anti-TNF withdrawal. In patients that relapsed, the comparison from baseline to point of relapse showed upregulation of key inflammatory genes, including IL1B, STAT3, OSMR, TNF and CXCL10. Pathway enrichment analysis showed that the upregulated genes were associated with an inflammatory response, including TNF-signaling. At two-year follow-up patients who remained in remission also showed upregulation of proinflammatory genes, including genes associated with TNF-signaling. However, the expression levels were lower than in the relapse group. Patients in the control group who experienced treatment failure (n=10) showed a gene expression signature that strongly overlapped with those that relapsed in the intervention group.

Conclusion

We did not detect any transcriptional differences between the relapse and remission group at baseline and after anti-TNF withdrawal the expression of proinflammatory genes including genes involved in TNF-signaling increased in both groups. However, a more pronounced increase was observed in the relapse group, suggesting that those that relapse exhibited reduced immunoregulatory capacity.

Introduction

Crohn ‘s disease (CD) is characterized by chronic inflammation in the gut, where severe complications such as fibrotic strictures require surgical resection. Stricture development involves excessive extracellular matrix (ECM) deposition due to continuous activation of intestinal myofibroblasts, that further contribute to intestinal fibrogenesis. Emerging data suggests that stricture-related intestinal myofibroblasts overexpress a serine protease called fibroblast activation protein (FAP). Furthermore, type III collagen deposition is increased during fibrosis in all layers of the intestinal tract, and studies have shown elevated collagen degradation in serum from patients with CD. The present study aimed at evaluating the potential of FAP-cleaved type III collagen [C3F] as a serum marker for intestinal fibrosis in CD.

Aims & Methods

The cohort consisted of 49 patients with luminal CD and 62 patients with stenotic CD. Clinical assessment was conducted for all patients at baseline and 12 months post study initiation. For the luminal phenotype, endoscopy was performed at both time points. For the stenotic group, radiographical assessment was done prior to surgery and endoscopy was performed at 12 months. C3F was measured in serum at all timepoints for baseline, post-operative day (POD), 1, 3, 6 and 12 months. To compare C3F levels at baseline for luminal vs stenotic phenotype, Mann-Whitney U-test was conducted. For the stenotic group, mixed-effect analysis, with a Dunnett’s multiple comparison applying Bonferroni Correction, was conducted to compare C3F levels for baseline vs POD and Spearman’s rank correlation was applied.

Results

C3F baseline levels were significantly higher in patients with stenotic CD compared to luminal CD (Table 1). In the stenotic group, C3F was significantly increased in patients at POD compared to baseline (Table 1). In the luminal group, C3F was significantly elevated in patients at 12 months compared to baseline. Furthermore, C3F showed a positive correlation with C-reactive protein (CRP) (ρ=0.350, p=0.006) and a significant negative correlation with disease duration (ρ= -0.257, p=0.048) for patients with the stenotic CD phenotype (Table 1).


Table 1. C3F levels are elevated in the stenotic CD vs the luminal CD group, with increased levels at POD compared to baseline for the stenotic phenotype. Spearman’s rank correlation, depicting the relationship between C3F and routine blood demographics, as well as demographics. Significant findings (p<0.05) are highlighted in bold.

Conclusion

C3F was elevated in patients with stenotic CD, with even higher levels after the surgical resection. The data suggests that C3F reflects fibroblast activity during fibrostenosis and tissue remodeling following surgery. The positive correlation with CRP supports the association with inflammation leading to collagen deposition and fibrosis. Interestingly, C3F shows negative correlation with disease duration suggesting that C3F could reflect fibrosis at early stages of stenotic CD. These findings highlight the potential use of C3F as a novel biomarker for intestinal fibrosis in CD.

Disclosure

TTK, MSA, MP are employed at Nordic Bioscience. ACBJ, JHM and MAK are employees and stock owners at Nordic Bioscience.

Introduction

In Crohn’s disease (CD), an exposure-response relationship is suggested for ustekinumab, with higher serum levels associated with better outcomes. Pharmacokinetic data are scarce, with suggested target maintenance trough level associated with remission at 0.8-1.4 µg/mL. ^{1, 2} The REScUE study (NCT04245215) investigated the effect of two different re-induction regimens with ustekinumab in patients with CD. No significant difference was observed on clinical, endoscopic and biological outcomes.³ Here we present the pharmacokinetic and pharmacodynamics results of the REScUE study.

Aims & Methods

This Belgian multicentre prospective double-blind randomized placebo-controlled trial included adult patients with CD treated with ustekinumab who presented with secondary loss of response to ustekinumab after documented primary response. All patients received a single IV re-induction with ustekinumab (≈6mg/kg) and were then randomized 1:1 to blinded maintenance ustekinumab 90 mg SC Q4W (intervention) or 90 mg SC Q8W (control) for 48 weeks. PK samples were obtained at baseline, week 4-8-12-20-28-36-48.

Results

In total, 108 patients were included: 67% female, median [IQR] age 41 [32-54] year, median disease duration 14 [7-22] years. Prior anti-TNF exposure was reported in 92% of the patients. Baseline ustekinumab trough levels at moment of loss of response were similar in both groups and at the suggested target level of 0.8-1.4 µg/mL ¹: median (IQR) q4 week 1.39 (0.84-2.74) µg/mL and q8 week 1.31 (0.91-2.34) µg/mL (p>0.9). Eight weeks after induction, there was a statistically significant difference in ustekinumab trough levels between both groups 10.8 (8.0-15.9) µg/mL versus 8.2 (4.9-11.1) µg/mL for q4weeks vs q8weeks respectively (p=0.002). After 48 weeks of treatment the statistically significant difference persisted with 6.0 (4.8-9.6) µg/mL versus 2.1 (1.6-2.9) µg/mL (p<0.001). No significant association between baseline/week 8/week 48 ustekinumab trough levels and (steroid free) clinical remisison [PRO2 (AP ≤ 1 AND SF ≤ 3) and FCP <250µg/g and no steroids for 90 days prior to week 48] or endoscopic remission [SES-CD <3 at week 48] was seen. However, there was a significant association between baseline/week 8/week 48 ustekinumab trough levels and biomarker remission [CRP <5 mg/L and FCP <250 µg/g at week 48]; OR (95% CI) 2.98 (1.85, 5.23), 1.12 (1.06, 1.19) and 1.35 (1.13, 1.66) (all p<0.01) respectively. As per quartile analysis of the ustekinumab trough levels at week 48, the lowest quartile (ustekinumab <0.9 µg/mL) had the lowest steroid free clinical remission in each treatment group.

Conclusion

In patients with CD experiencing secondary loss of response to ustekinumab, baseline ustekinumab levels are low. Dose optimization with a single IV re-induction followed by intensified maintenance dosing (90 mg SC q4W) leads to higher ustekinumab levels compared to IV re-induction followed by conventional maintenance dosing (90 mg SC q8W). No association was found between ustekinumab levels and different outcomes except for biomarker remission. This suggests that treatment with ustekinumab should be personalized on a patient level, guided by drug-clearance rather than guided by target trough levels.

References

¹ Adedokun O. et al Gastroenterology 2018;154:1660–1671
² Schreiber S. et al ECCO 2023 Copenhagen; P436
³ Bossuyt P. et al ECCO 2025 Berlin; OP35

Disclosure

PB: Financial support for research: AbbVie, EG. •Lecture fees: AbbVie, AMC ICP, Amgen, Bristol Myers Squibb, Celltrion, Dr Falk Benelux, EG, Galapagos, Globalport, Lilly, Medtalks, Materia Prima, Pentax, Springer Media. •Advisory board: AbbVie, Bristol Meyers Squibb, CIRC, Galapagos, Janssen, Lilly, Pentax, PSI-CRO, Roche, Takeda, Tetrameros.

Introduction

The Mayo Endoscopic Subscore (MES) is widely used in clinical trials and daily practice to assess disease activity and response to treatment in UC.

Aims & Methods

The objective of this study was to identify any nuances in patients with MES > 1 treated with non-TNF biologics and targeted oral therapies. The Ipsos UC Therapy Monitor, a multi-centre online medical chart review study of patients with UC, was conducted between July and September 2024 among gastroenterologists practicing across hospital and private practices in the UK, France, Germany, Italy, and Spain. Physicians were screened for practice duration and patient volume. Charts of patients prescribed advanced therapy were included in the analysis.

Results

205 sampled physicians collectively reported 820 UC patients. Of the reported patients with MES > 1, 62 were treated with non-TNF biologics and 62 were treated with novel oral therapies. Among the patients treated with non-TNF biologics, a higher proportion were female vs those receiving oral therapy (48% vs 31%); they were also older in age (mean years: 43 vs 38).
The patients treated with non-TNF biologics had a higher prevalence of comorbidities, including depression/anxiety (24% vs 19%), cardiovascular disease (8% vs 0%), and dyslipidemia/hyperlipidemia/hypercholesterolemia (11% vs 7%), compared to those treated with novel oral therapy.
Mean CRP levels were higher in the oral therapy group (10.2 mg/L vs 9.1 mg/L), while mean Mayo scores were slightly lower (4.7 vs 5.2). Fecal calprotectin levels were elevated in patients treated with non-TNF biologics vs oral therapy group (441 µg/g vs 503 µg/g).
When analysing reasons for prescribing selected advanced therapy between the two patient groups, a greater proportion of patients treated with non-TNF biologics had safety reasons specified more frequently by the sampled physicians.

Table 1: % of reported UC patients receiving advanced therapy for specific cited reasons

Conclusion

This study highlights key differences in patient characteristics and treatment considerations among UC patients with MES > 1 treated with non-TNF biologics compared to those treated with targeted oral therapies. Patients on non-TNF biologics tend to be older, more often female, and have a higher prevalence of comorbidities. Physicians prescribing non-TNF biologics prioritize safety, while those prescribing oral therapies focus on rapid symptom control. These findings highlight the importance of considering patient characteristics and treatment goals when selecting advanced therapies for UC patients with MES > 1. Further investigation using comparator cohort is warranted.

Introduction

The pathogenesis of ulcerative colitis (UC) involves complex immune responses, including Th1, Th2, and Th17 pathways, and is characterized by substantial heterogeneity. Cytokine profiling may contribute to understanding individual disease mechanisms and guiding personalized therapeutic strategies; however, its clinical application remains limited due to technical complexity. This study aimed to investigate whether endocytoscopic and histopathological findings correlate with mucosal cytokine expression levels and could serve as surrogate markers.

Aims & Methods

Thirty-five patients with UC receiving treatment at our institution were enrolled. Endocytoscopy (ultra-magnifying endoscopy) was performed in all patients, and the findings were classified according to the Endocytoscopic Classification system (EC classification: EC-A, EC-B, EC-C, EC-D). Mucosal biopsies were obtained from the most inflamed areas identified during endocytoscopy. Histological evaluation included the Matts histological grade and assessments of neutrophil, eosinophil, and lymphocyte infiltration; goblet cell depletion; crypt distortion; and epithelial damage, each graded on a four-point scale (absent, mild, moderate, severe). mRNA expression levels of cytokines (IFN-γ, TGF-β1, TNF-α, IL-1β, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, IL-13, IL-17, and IL-23) were quantified using RT-PCR. Statistical analyses were performed to assess correlations between endocytoscopic/histopathological findings and cytokine expression levels.

Results

The median age of participants was 44 years (IQR: 35–57), with 24 males and 11 females. Disease types included pancolitis (n = 16), left-sided colitis (n = 14), and proctitis (n = 5). The median Mayo score was 1 (IQR: 0–5). EC classification scores showed significant positive correlations with the expression levels of IL-1β, IL-6, IL-8, IL-13, IL-17, and IL-23 (Spearman’s rank correlation, p < 0.05; r = 0.555, 0.526, 0.584, 0.441, 0.422, and 0.449, respectively). Similarly, Matts histological grades were significantly correlated with the expression of IFN-γ, IL-1β, IL-6, IL-8, IL-13, IL-17, and IL-23 (p < 0.05; r = 0.493, 0.756, 0.692, 0.730, 0.656, 0.623, and 0.448, respectively). Histopathological features—including neutrophil and lymphocyte infiltration, goblet cell depletion, epithelial damage, and crypt distortion—were significantly associated with the expression of IFN-γ, IL-1β, IL-6, IL-8, IL-13, IL-17, and IL-23. Additionally, eosinophil infiltration was positively correlated with the expression levels of IL-1β, IL-5, and IL-13.

Conclusion

This study demonstrates that endocytoscopic and histopathological indicators of inflammation are associated with mucosal expression levels of several cytokines—particularly inflammatory and Th17-related cytokines—in patients with ulcerative colitis (UC). These findings suggest that such evaluations may serve as surrogate markers, potentially improving disease assessment and informing therapeutic decision-making.

Introduction

The Mayo Endoscopic Subscore (MES) is the most widely used endoscopic index for assessing disease activity in ulcerative colitis (UC). Although artificial-intelligence　(AI)–assisted colonoscopy is expected to reduce inter-observer variability, the MES 4-point scale (0–3) lacks sufficient sensitivity to detect subtle changes in mucosal activity, limiting its ability to capture incremental improvement or deterioration. Moreover, because MES focuses on qualitative features—such as bleeding and erosions—without considering the extent or surface area of inflammation, it provides an incomplete estimate of a patient’s overall inflammatory burden.

Aims & Methods

We sought to develop and validate a quantitative, AI–driven colonoscopic score—Quantitative Ulcerative Colitis Assessment using Deep-learning (QUAD)—and to determine whether it predicts clinical relapse in patients with UC. The QUAD system was trained on 84 743 colonoscopic images from 998 UC patients at three centers and comprises three main functions: i) Image filtering: Automatic exclusion of low-quality images that are unsuitable for analysis owing to light saturation (halation), inadequate insufflation, excessive proximity to the mucosa, chromoendoscopy, or virtual chromoendoscopy. ii) Frame-level scoring: Each frame is divided into four quadrants; inflammation in each quadrant is graded from 0 to 3, and the quadrant scores are summed to yield a QUAD score of 0–12. iii) Video-level aggregation: Full-length withdrawal videos are processed to generate (i) an Entire QUAD score (0–60), representing pan-colonic inflammation, and (ii) a Distal QUAD score (0–12) for the rectosigmoid segment (final 20 % of the video).
The QUAD system contains mainly three function: i) automatically exclude low quality images that is ineligible for analysis in cases of halation, inadequate air de­livery, proximity to mucosa, chromoendoscopy, and virtual chromoendoscopy; ii) fully auto­mated twelve-class score output based on frame analysis: Each frame was divided into four quadrants; inflammation in each quadrant was graded 0–3 and summed to yield a QUAD score of 0–12; and iii) full-length withdrawal videos were processed to generate (I) an Entire QUAD (0–60) for considering whole-colon inflammation status and (II) a Distal QUAD (0–12) for the distal rectum–sigmoid segment (final 20 % of the video). An open-label cohort of adults with quiescent UC (partial Mayo score ≤ 2 for ≥6 months) was prospectively enrolled at a single center (August 2022–February 2023), and followed for 24 months after baseline colonoscopy; clinical relapse was defined as partial Mayo score > 2. The predictive value of QUAD scores for relapse was evaluated.

Results

Frame-level QUAD scores correlated strongly with expert-assigned MES values (Spearman r = 0.68, p < 0.01). Among 156 participants (median age, 51 years; 49 % male), 22 experienced relapse during follow-up. Relapse occurred in 19.4 % (19/98) of patients with a QUAD score > 3 versus 5.2 % (3/58) with a score ≤ 3 (log-rank p = 0.01).
In video analysis, an Entire QUAD > 6 did not significantly stratify relapse risk (16.8 % [17/101] vs 9.1 % [5/55]; log-rank p = 0.17). In contrast, a Distal QUAD > 3 was strongly associated with relapse (43.8 % [7/16] vs 10.7 % [15/140]; log-rank p < 0.01).

Conclusion

AI-generated QUAD provides a sensitive, fully quantitative measure of mucosal inflammation and stratifies relapse risk in clinically quiescent UC. In particular, the automatically derived Distal QUAD—focused on the distal colon and independent of operator image selection—offers a practical, objective tool for longitudinal monitoring and treatment tailoring in UC.

Introduction

Crohn’s disease (CD) is a human immune-mediated inflammatory disease (IMID) of unknown etiology that affects the gastrointestinal (GI) tract. CD is now a global health issue due to its lifelong nature and significant burden on patients and public health [1]. Phase III clinical trials have shown that treatment with oral Janus Kinase (JAK) inhibitors can alleviate disease in patients with CD. However, the United States (US) Food and Drug Administration (FDA) has issued warnings about side effects for this class of oral drugs which target JAKs systemically in the human body. Therefore, new targeted delivery approaches are needed to block JAK activity selectively in the GI tract. The JAK1/2-STAT1 intracellular signaling pathway is activated by the Th1 cytokine IFN-γ alone or in synergy with another Th1 cytokine, TNF-α, and SOSC1 is an endogenous negative regulator of JAK1/2. This pathway activates the expression of inflammatory genes in multiple cell types typically associated with active or inflamed CD [2]. RNA interference (RNAi) mediated gene silencing using small interfering RNA (siRNA) targeting JAK1/2-STAT1 and SOCS1 overexpression using exogenous SOCS1 mRNA are promising methods to reduce the JAK-STAT pathway activation.

Aims & Methods

The GENEGUT project aims to develop oral RNA-based therapies for ileal CD. In this study, we have developed new approaches to deliver siRNAs targeting JAK1, JAK2, and STAT1 into relevant cell models, including human intestinal epithelial cell (IEC) lines. Epithelial cell models that replicate the inflammatory microenvironment in CD were used by exposing IECs to inflammatory cytokines IFN- γ/TNF- α, leading to increased cytokine production and cell death.

Results

Selected siRNAs effectively blocked JAK1, JAK2 and STAT1 expression and activity, preventing the downstream activation of inflammatory and cell death pathways. SOCS1 overexpression also suppressed JAK1/2-STAT1 pathway activation and enhanced cell survival.

Conclusion

Ongoing work is focused on optimizing the delivery of siRNA and mRNA into differentiated epithelial monolayers, co-cultures and patient-derived organoids to advance translational applications.

References

1. Dulai PS, Singh S, Vande Casteele N, Boland BS, Rivera-Nieves J, Ernst PB, Eckmann L, Barrett KE, Chang JT, Sandborn WJ. Should We Divide Crohn's Disease Into Ileum-Dominant and Isolated Colonic Diseases? Clin Gastroenterol Hepatol. 2019 Dec;17(13):2634-2643. doi: 10.1016/j.cgh.2019.04.040. Epub 2019 Apr 19. PMID: 31009791; PMCID: PMC6885453.
2. Cordes F, Foell D, Ding JN, Varga G, Bettenworth D. Differential regulation of JAK/STAT-signaling in patients with ulcerative colitis and Crohn's disease. World J Gastroenterol. 2020 Jul 28;26(28):4055-4075. doi: 10.3748/wjg.v26.i28.4055. PMID: 32821070; PMCID: PMC7403801.

Disclosure

Funded by the European Union (GA 101057491) and supported the Swiss State Secretariat for Education, Research and Innovation (SERI) under contract number 22.00119. Views and opinions expressed are however those of the author(s) only and do not necessarily reflect those of the European Union or the European Health and Digital Executive Agency (HaDEA). Neither the European Union nor the granting authority can be held responsible for them. UK participants in Horizon Europe Project GENEGUT are supported by UKRI grant number 10042451 (Bangor University).