Introduction

Metabolic dysfunction-associated steatohepatitis (MASH) increases the risk of cardiovascular (CV) events and liver fibrosis. Semaglutide, a glucagon-like peptide-1 (GLP-1) receptor agonist, reduces CV risk associated with overweight and obesity in the absence of diabetes and has a profound effect on weight reduction and MASH resolution. Management of MASH may benefit from a deeper understanding of the mechanistic basis of semaglutide-induced MASH resolution.

Aims & Methods

This study used a multi-omics approach to investigate the effects of semaglutide on the hepatic microenvironment and fibrosis development in the context of semaglutide-induced MASH resolution. GLP-1 receptor expression was assessed using RNA in situ hybridization and immunohistochemistry in human liver tissue and immunohistochemistry in liver tissue from DIO‑NASH and CDA-HFD mice. The effect of semaglutide treatment on liver fibrosis was also assessed using liver transcriptome analysis in both of these mouse models of MASH. Proteomic analyses of 4,979 proteins using the SomaScan Platform were conducted on serum samples from 293 patients with MASH who received semaglutide or placebo in a phase 2b study (NCT02970942), and an independent cohort of 141 patients with MASH and 89 healthy volunteers.

Results

GLP-1 receptor mRNA or protein expression were not detected in human or mouse liver tissue. However, semaglutide improved histological markers of fibrosis and expression of genes involved in collagen turnover in two distinct mouse models of MASH; one that phenocopies the metabolic-associated features of human disease and another that exhibits non-abdominal obesity-induced advanced steatohepatitis and rapid fibrosis. In semaglutide-treated patients with MASH, proteomics identified 72 proteins significantly associated with MASH resolution after 72 weeks of treatment. These 72 proteins were also identified as differentially expressed in patients with MASH who were not treated for MASH, relative to healthy individuals. In semaglutide-treated patients with MASH, the abundance of the 72 proteins was similarly altered; however, semaglutide reverted the altered disease profile to that seen in non-pharmacologically treated healthy individuals. In contrast, changes in the levels of the 72 proteins in patients who received placebo were negligible. Notably, changes in the abundance of 26 of the 72 proteins were significant even after correction for weight loss. Although most of these 26 proteins are associated with metabolic functions, several are implicated in fibrosis-related pathways and CV disease (e.g. ADAMTSL2 and ACY1), and hepatocellular carcinoma (e.g. SERPINC1).

Conclusion

Based on circulating proteomic profiling, semaglutide-induced MASH resolution is associated with changes in multiple pathogenic pathways even after correction for weight loss. Semaglutide appears to re-balance the expression levels of disease-associated proteins in MASH to levels found in healthy individuals.

Disclosure

MJ is an employee of Novo Nordisk A/S. JN is an employee of Novo Nordisk A/S. MSK is an employee of Novo Nordisk A/S. KA is an employee of Novo Nordisk A/S. KMB is an employee of Novo Nordisk A/S. EB served as a consultant or advisory board member for Boehringer Ingelheim, Gilead Sciences, Intercept, Merck, Novo Nordisk, Pfizer, ProSciento. Grants from Gilead Sciences for fatty liver research, Speaker for Gilead Sciences, Intercept, Merck, Novo Nordisk, and Pfizer. KC served as a consultant for Aligos Therapeutics, Arrowhead, AstraZeneca, 89Bio, BMS, Eli Lilly & Co., Merck, Novo Nordisk, Prosciento, Sagimet Biosciences, Siemens, and Terns Pharma. Received research support (University of Florida; Principal Investigator) from Boehringer Ingelheim, Echosens, Inventiva, Labcorp, and Perspectum. EDG is an employee of Novo Nordisk A/S. MG is an employee of Novo Nordisk A/S. LLG is an employee of Novo Nordisk A/S. LMH is an employee of Novo Nordisk A/S. RL is a consultant to Aardvark Therapeutics, Altimmune, Anylam/Regeneron, Amgen, Arrowhead Pharmaceuticals, AstraZeneca, Bristol-Myer Squibb, CohBar, Eli Lilly, Galmed, Gilead, Glympse bio, Hightide, Inipharma, Intercept, Inventiva, Ionis, Janssen Inc., Madrigal, Metacrine, Inc., NGM Biopharmaceuticals, Novartis, Novo Nordisk, Merck, Pfizer, Sagimet, Theratechnologies, 89 bio, Terns Pharmaceuticals and Viking Therapeutics. His institutions received research grants from Arrowhead Pharmaceuticals, Astrazeneca, Boehringer-Ingelheim, Bristol-Myers Squibb, Eli Lilly, Galectin Therapeutics, Galmed Pharmaceuticals, Gilead, Hanmi, Intercept, Inventiva, Ionis, Janssen, Madrigal Pharmaceuticals, Merck, NGM Biopharmaceuticals, Novo Nordisk, Pfizer, Sonic Incytes and Terns Pharmaceuticals. Co-founder of LipoNexus Inc. GM is an employee of Novo Nordisk A/S. PNN is a consultant for Novo Nordisk, Boehringer Ingelheim, Gilead, Intercept, Poxel Pharmaceuticals, Bristol Myers Squibb (BMS), Pfizer, Sun Pharma, Madrigal and GSK. He also reports honoraria as a speaker for Novo Nordisk and AiCME; support for attending meetings from for Novo Nordisk; and participation on an advisory board for Novo Nordisk, Boehringer Ingelheim, Gilead, Intercept, Poxel Pharmaceuticals, BMS, Pfizer, Sun Pharma, Madrigal, and GSK. LMN is an employee of Novo Nordisk A/S. MSP is an employee of Novo Nordisk A/S. VR received consulting fees from Novo-Nordisk, Sagimet, Madrigal, Enyo, Poxel, Northsea, Intercept Pharmaceuticals, Prosciento. Research grants (to institution) from Gilead Sciences and Intercept Pharmaceuticals. A-SS is an employee of Novo Nordisk A/S. VWW served as a consultant or advisory board member for AbbVie, Boehringer Ingelheim, Echosens, Gilead Sciences, Intercept, Inventiva, Novo Nordisk, Pfizer, Sagimet Biosciences, and TARGET PharmaSolutions. Has received a research grant from Gilead Sciences. Speaker for Abbott, AbbVie, Gilead Sciences, Novo Nordisk, and Unilab. Co-founder of Illuminatio Medical Technology Limited. Has received travel support from AbbVie and Gilead Sciences. QMA is coordinator of the IMI2 Liver Investigation: Testing Marker Utility in Steatohepatitis (LITMUS) consortium, which is funded by the European Union Horizon 2020 programme and the European Federation of Pharmaceutical Industries and Associations (EFPIA). This multi-stakeholder consortium includes industry partners. He reports research grant funding from AstraZeneca, Boehringer Ingelheim, and Intercept; consultancy on behalf of Newcastle University for Alimentiv, Akero, AstraZeneca, Axcella, 89Bio, Boehringer Ingelheim, Bristol Myers Squibb, Galmed, Genfit, Genentech, Gilead, GlaxoSmithKline, Hanmi, HistoIndex, Intercept, Inventiva, Ionis, IQVIA, Janssen, Madrigal, Medpace, Merck, NGMBio, Novartis, Novo Nordisk, PathAI, Pfizer, Prosciento, Poxel, Resolution Therapeutics, Roche, Ridgeline Therapeutics, RTI, Shionogi, and Terns; speaker fees/honoraria from Fishawack, Integritas Communications, Kenes, Novo Nordisk, Madrigal, Medscape, and Springer Healthcare; and royalties from Elsevier. LBK is an employee of Novo Nordisk A/S.

Introduction

Irritable Bowel Syndrome (IBS) is a prevalent and chronic gastrointestinal disorder that affects approximately 85 million Europeans. It predominantly impacts women and significantly impairs quality of life, presenting with recurrent abdominal pain and altered bowel habits in the absence of identifiable structural abnormalities. Beyond its gastrointestinal symptoms, IBS frequently co-occurs with mental and somatic conditions such as anxiety, depression, fibromyalgia (32.5%), and chronic fatigue syndrome (14%). Up to 60% of IBS patients experience mental comorbidities, highlighting the complex interplay between gut and brain—often referred to as dysregulation of the gut–brain axis. This dysregulation manifests in altered intestinal motility, increased gut permeability, visceral hypersensitivity, and significant shifts in gut microbiota composition.

Aims & Methods

We present the DISCOVERIE project (Development, dIagnostic and prevention of gender-related Somatic and mental COmorbitiEs in iRritable bowel syndrome In Europe), an international, multi-centre initiative aimed at characterizing IBS and its associated comorbidities, with a particular focus on sex- and gender-related differences. To this end, we generated a comprehensive multi-omics dataset from 743 individuals, including shotgun quantitative microbiome profiling and untargeted metabolomics. These molecular data were complemented by in-depth clinical phenotyping, capturing information on anxiety, depression, fibromyalgia, and chronic fatigue syndrome. Particular attention was given to age, sex/gender, lifestyle factors, and systemic inflammatory markers. Multivariate statistical models were applied, correcting for common microbiome confounders, to analyze the multi-omics layers and identify microbiome-related signatures and mechanistic links across IBS and its comorbid subgroups.

Results

Our microbiome analysis revealed that IBS subjects exhibit features of a dysbiotic bacterial community, characterized by an increased prevalence of the Bacteroides 2 enterotype (17–26% in IBS participants across comorbid groups vs. 17% in the general population and only 2% in the healthy control group; chi-squared test, FDR < 0.05), a reduced butyrate synthesis capacity, and increased levels of opportunistic taxa compared to non-IBS controls.
Interestingly, IBS individuals—regardless of comorbidities—showed enrichment of bacterial metabolic modules involved in the degradation of amino acids that serve as precursors for neurotransmitters. Specifically, we observed an increase in pathways associated with GABA degradation and a decrease in modules responsible for glutamate biosynthesis. This dysregulation of gut neurotransmitter metabolism may contribute to hallmark IBS symptoms.
Strikingly, pro-inflammatory IL-6 levels were strongly associated with specific bacterial genus (FDR < 0.05), as well as with a metabolic module responsible for the degradation of gamma-hydroxybutyric acid (GHB), a neurotransmitter and central nervous system depressant, suggesting a potential mechanistic link between low-grade inflammation and depressive symptoms in IBS.

Conclusion

The results from this large-scale, multi-center international study reveal a dysregulation of gut neurotransmitter metabolism that may contribute to IBS symptoms and associated comorbidities. These findings underscore a potential link between elevated IL-6 levels, microbial metabolism, and the gut–brain axis, offering new insights into the complex interplay between microbiota composition, host physiology, and IBS-related comorbidities.

Introduction

Colorectal cancer (CRC) often develops from precancerous lesions such as tubular adenomas (TA) and sessile serrated lesions (SSL). The gut microbiome has been implicated in colorectal carcinogenesis, but specific microbial signatures associated with different adenoma subtypes remain incompletely characterized. This study aimed to identify microbial biomarkers associated with TA and SSL, and explore their correlations with dietary macronutrients.

Aims & Methods

Stool samples were collected from 801 patients prior to colonoscopy from National University Hospital and Tan Tock Seng Hospital, Singapore. After colonoscopy, the patients were classified as having TA (n=315), SSL (n=24), both (n=8), or neither (controls, n=454). Shotgun metagenomic sequencing was performed using Novaseq6000, and microbial taxonomy was annotated with MetaPhlAn4. Multivariate linear regression adjusted for age, gender, and BMI was used to identify species associated with polyp subtypes. A subset of 172 participants completed a validated food frequency questionnaire (FFQ) assessing 19 macronutrients, enabling correlation analysis between diet and adenoma-associated microbes.

Results

Patients with TA were older (mean age 64.0±7.8 vs 62.3±7.5 years in controls, p=0.02) and more likely to be male (69.5% vs 52.4%, p<0.01). After adjustment for confounders, we identified 38 microbial species significantly associated with polyp subtypes; 20 were associated with SSL and 18 with TA, with minimal overlap between the two polyp subtypes. Only two species showed associations with both polyp types: Holdemanella biformis and GGB3277_SGB4327, with the latter showing opposing directional relationships (negative for TA, positive for SSL). Of note, SSL were associated with increased abundance of Faecalibacterium SGB15346 and Bifidobacterium adolescentis. In the dietary analysis, we identified significant correlations between SSL-associated microbes and sugar intake, particularly Faecalibacterium SGB15346 (r=0.170, p=0.03) and Clostridium sp_AM49_4BH (r=0.156, p=0.04). E. hominis, positively associated with SSL, showed the strongest dietary associations overall, correlating positively with carbohydrate intake (r=0.243, p<0.01) and starch consumption (r=0.246, p<0.01).

Conclusion

Our study identified distinct microbial signatures associated with TA and SSL, with minimal overlap, suggesting separate host-microbial biological pathways in their development. The positive correlation between SSL-associated microbes and sugar intake provides a potential mechanistic link between dietary habits and SSL formation, which may be particularly relevant given the increasing incidence of early-onset colorectal neoplasia. These findings offer potential microbial biomarkers for early detection of colorectal polyps and suggest targeted dietary interventions as a strategy for precision chemoprevention.

Introduction

Anti-tumor necrosis factor (TNF)-α are the first target therapies approved for the treatment of steroid-dependent or -refractory ulcerative colitis (UC). Drugs with different mechanisms of action have recently entered the market, but data on the optimal management of UC after failure of first-line anti-TNF-α are lacking.

Aims & Methods

The PODIUM study is a multicentre, retrospective, observational study that aims to compare the real-life effectiveness and safety of second-line vedolizumab (VDZ), ustekinumab (UST) and tofacitinib (TOF) in patients with UC. Consecutive patients with UC who had failed at least one anti-TNF-α and were never exposed to other target therapies were enrolled from 12 centres across Europe. Sociodemographic data, data on clinical activity and adverse events of interest (AEIs, defined as AE requiring temporary/permanent drug discontinuation or medical intervention) were retrospectively extracted from medical records, from the initiation of second-line therapy until one year of observation/drug discontinuation/last available follow-up. The primary outcome was the cumulative probability of achieving steroid-free clinical remission (SFCR), defined as stool frequency=0-1 and rectal bleeding=0 and no concomitant steroids, over the first year of treatment; inverse probability of treatment weighting (IPTW), calculated as the inverse of the probability of receiving each drug (IPTW=1/Propensity Score), was applied to the primary outcome to account for imbalances in cohorts’ characteristics. We performed an interim analysis including all data collected up to March 15^th 2024; data collection is still ongoing at the submission of this Abstract.

Results

Data on 303 patients (172 VDZ, 63 UST, 68 TOF) were included in the analysis. The main baseline characteristics are presented in Table 1. After IPTW, the cumulative probability of achieving SFCR during the first year was 51.0%, 59.1% and 64.1% for VDZ, UST and TOF, respectively; both UST and TOF were superior to VDZ at Log-rank test (p<0.001), with no differences between the two. At weighted logistic regression, female sex (adjusted Odds Ratio, aOR, 0.06, Confidence Interval, CI, 95% 0.41-0.73, p<0.001) and higher Charlosn Comorbidity Index calculated without considering age (aOR 0.73, 95% CI 95% 0.60-0.88, p=0.001) were independently associated with a lower probability of achieving SFCR during the first year. Recorded AEIs were 16 (4 infections) in the VDZ group (9.3% of VDZ-treated patients), 6 (1 infection) in the UST group (9.5% of UST-treated patients), and 15 (5 infections, including 1 Zoster reactivation) in the TOF group (22.1% of TOF-treated patients).
Table 1

Conclusion

This preliminary analysis suggests that both UST and TOF might have superior effectiveness, compared to VDZ, as second-line agents in patients with UC after anti-TNF-α. Confirmatory analysis with a larger sample size will be performed. A slightly higher rate of AEIs was observed in the TOF group.

Disclosure

Giuseppe Privitera received consultancy fees from Alphasigma and Janssen. Cristina Bezzio received lecture fees and served as a consultant for Takeda, MSD, Ferring, Abbvie, Galapagos and Janssen. Edoardo Vincenzo Savarino has served as speaker for Abbvie, Agave, AGPharma, Alfasigma, Aurora Pharma, CaDiGroup, Celltrion, Dr. Falk, EG Stada Group, Fenix Pharma, Fresenius Kabi, Galapagos, Janssen, JB Pharmaceuticals, Innovamedica/Adacyte, Malesci, MayolyBiohealth, Omega Pharma, Pfizer, Reckitt Benckiser, Sandoz, SILA, Sofar, Takeda, Tillots, and Unifarco; has served as a consultant for Abbvie, Agave, Alfasigma, Biogen, Bristol-Myers Squibb, Celltrion, DiademaFarmaceutici, Dr. Falk, Fenix Pharma, Fresenius Kabi, Janssen, JB Pharmaceuticals, Merck & Co, Nestlè, Reckitt Benckiser, Regeneron, Sanofi, SILA, Sofar, Synformulas GmbH, Takeda, and Unifarco; he has received research support from Pfizer, Reckitt Benckiser, SILA, Sofar, Unifarco, Zeta Farmaceutici. Daniela Pugliese received speaker fees and/or advisory board from AbbVie, Galapagos, MSD, Takeda and Janssen, Pfizer. Iago Rodriguez-Lago has received financial support for traveling and educational activities from or has served as an advisory board member for Abbvie, Adacyte, Celltrion, Chiesi, Danone, Ferring, Faes Farma, Janssen, Galapagos, MSD, Pfizer, Roche, Takeda, and Tillotts Pharma. Financial support for research: Tillotts Pharma. Manuel Barreiro-de Acosta has served as a speaker, consultant and advisory member for and has received research funding from MSD, AbbVie, Janssen, Kern Pharma, Celltrion, Takeda, Gilead, Celgene, Pfizer, Sandoz, Biogen, Fresenius, Ferring, Faes Farma, Dr. Falk Pharma, Chiesi, Gebro Pharma, Adacyte and Vifor Pharma. Sara Onali has received speaker fees from AbbVie, Takeda, Janssen, Amgen, Galapagos and Norgine. Massimo Claudio Fantini has received speaker and consultancy fees from AbbVie, Takeda, Janssen-Cilag, Pfizer, Sandoz, Biogen, Galapagos, Bristol-Myers Squibb, and MSD; and research grants from Pfizer and Janssen-Cilag. Karmiris K has served as speaker, consultant, and advisory member for Abbvie, Amgen, Enorasis, Ferring, Galenica, Genesis, Janssen, MSD, Pfizer, Takeda and Vianex. Henit Yanai has received consulting fees Abbvie, Ferring, Janssen, Neopharm Ltd., Pfizer, Takeda; Payment or honoraria for lectures, presentations, speakers bureaus, manuscript writing or educational events Abbvie, Janssen, Pfizer, Takeda, Neopharm Ltd.; Participation on a Data Safety Monitoring Board or Advisory Board Abbvie, Neopharm Ltd., Pfizer, Takeda. Ana Gutierrez has served as speaker, consultant and advisory member for or has received research funding from MSD, Abbvie, Pfizer, Kern Pharma, Takeda, Janssen, Ferring, Faes Farma, Shire Pharmaceuticals, Tillotts Pharma, Chiesi and Otsuka Pharmaceutical. Luca Pastorelli has received consultant and/or speaker fees from Giuliani, Ferring, Janssen, Takeda. María Chaparro has served as a speaker and consultant for and has received research or education funding from MSD, AbbVie, Hospira, Pfizer, Takeda, Janssen, Ferring, Shire Pharmaceuticals, Dr. Falk Pharma, Tillotts Pharma, Biogen, Gilead and Lilly. Yamile Zabana has received speaker and/or consultant fees from AbbVie, Adacyte, Almirall, Amgen, Dr Falk Pharma, FAES Pharma, Ferring, Janssen, MSD, Otsuka, Pfizer, Shire, Takeda, Galapagos, Boehringer Ingelheim and Tillots. Alessandro Armuzzi: consulting and/or advisory board fees from AbbVie, Amgen, Biogen, Boehringer Ingelheim, Bristol-Myers Squibb, Celgene, Celltrion, Eli-Lilly, Ferring, Galapagos, Gilead, Janssen, MSD, Mylan, Pfizer, Protagonist Therapeutics, Roche, Samsung Bioepis, Sandoz, Takeda; lecture and/or speaker bureau fees from AbbVie, Amgen, Arena, Biogen, Bristol-Myers Squibb, Eli-Lilly, Ferring, Galapagos, Gilead, Janssen, MSD, Mitsubishi-Tanabe, Novartis, Pfizer, Roche, Sandoz, Samsung Bioepis, Takeda; and research grants from MSD, Pfizer, Takeda and Biogen. The remaining authors declare no competing interests. The remaining Authors declare no competing interests.