Introduction

Inflammatory bowel diseases (IBD) have an ongoing, unmet medical need for new medicinal products with a different mode of action and a favourable safety profile. MH002 is a live biotherapeutic product (LBP) consisting of 6 commensal bacteria and is entering phase 3 clinical development as a therapeutic treatment developed to restore dysbiosis and depletion of butyric acid producing-bacteria that are associated with IBD. The non-pathogenic nature of the MH002 strains and absence of systemic bioavailability are expected to result in a particularly favourable safety profile.

Aims & Methods

To assess the safety of MH002 in ulcerative colitis (UC) and pouchitis (PC).
Safety data from 45 UC patients randomized (2:1) to double-blind treatment with 400mg MH002 or placebo daily for up to 16 weeks (MH002-UC-201), and from 14 PC patients treated open label with 400mg MH002 daily for 8 weeks (MH002-PC-201) were collected prospectively, the primary endpoint being occurrence of treatment emergent adverse events (AEs). UC patients were on stable 5-ASA treatment with or without low-dose corticosteroid throughout the study. Concomitant and recent antibiotic use was prohibited in all, as was treatment with advanced therapies. Placebo-treated UC patients who did not have a clinical response after 8 weeks were switched to MH002 treatment for 8 weeks.

Results

A total of 52 patients were exposed to MH002 with a total of about 136 patient-months of exposure. At baseline, the mean (range) modified Mayo score in UC patients was 5.76 (4.0-8.0) and the modified mean (range) Pouchitis Disease Activity Index score in PC patients was 6.91 (5.3-9.3). In UC patients, 20% were on stable, low dose corticosteroid, none was in the PC study. In both studies, no deaths, no severe AEs, no related serious AEs were reported in MH002-treated subjects, and no discontinuation occurred for an AE in MH002-treated participants. In particular, no bacteremia or sepsis was reported. Most AEs were mild and unrelated. Numerically less patients experienced an AE with MH002 than with placebo in UC.

	MH002-UC-201		MH002-PC-201	Total
Nr (%) of subjects	MH002 (n=31)	Placebo (n=14)	MH002 (n=14)	MH002 (n=45)​
with a severe Adverse Event (AE)	0	0	0	0
discontinuing for an AE	0	1 (7%)	0	0
with at least 1 AE	11 (35%)	8 (57%)	11 (79%)	22 (49%)
with a Serious AE (not related)	1 (3%)	1 (7%)	1 (7%)	2 (4%)
with a possibly related AE	1 (3%)	1 (7%)	3 (21%)	4 (9%)
with an AE of gastro-intestinal disorders	2 (6%)	4 (29%)	4 (29%)	6 (13%)

Possibly related AEs reported with MH002 included rash (UC) and abdominal pain, diarrhoea, flatulence and nausea (PC), while abdominal distension was reported with placebo (UC). Type 2 diabetes, arterial hypertension (MH002) and Clostridioides difficile infection (placebo) were the unrelated serious AEs that were reported. No clinically significant changes were observed during physical examination, vital signs assessment and in routine laboratory safety analyses (biochemistry, hematology and urinalysis) with MH002 or placebo. Efficacy data showed positive trends for clinical and endoscopic improvement and have been reported previously.

Conclusion

MH002 was well tolerated and safe in the first-in-disease phase 2 clinical trials conducted in UC and PC. MH002 may exhibit a particularly favourable safety profile if these results are confirmed in larger, late-stage clinical studies.

Disclosure

SV: financial support for research from AbbVie, J&J, Pfizer, Takeda and Galapagos; speakers’ and/or consultancy fees from AbbVie, Abivax, AbolerIS Pharma, AgomAb, Alimentiv, Arena Pharmaceuticals, AstraZeneca, BMS, Boehringer Ingelheim, Celgene, Cytoki Pharma, Dr Falk Pharma, Ferring, Galapagos, Genentech-Roche, Gilead, GSK, Hospira, Imidomics, Janssen, J&J, Lilly, Materia Prima, Mestag Therapeutics, MiroBio, Morphic, MRM Health, Mundipharma, MSD, Pfizer, Prodigest, Progenity, Prometheus, Robarts Clinical Trials, Surrozen, Takeda, Theravance, Tillots Pharma AG, VectivBio, Ventyx, Zealand Pharma.
SD: consultancy fees from AbbVie, Alimentiv, Allergan, Amgen, Applied Molecular Transport, AstraZeneca, Athos Therapeutics, Biogen, Boehringer Ingelheim, Bristol Myers Squibb, Celgene, Celltrion, Dr Falk Pharma, Eli Lilly, Enthera, Ferring Pharmaceuticals Inc., Gilead, Hospira, Inotrem, Janssen, Johnson & Johnson, Morphic, MSD, Mundipharma, Mylan, Pfizer, Roche, Sandoz, Sublimity Therapeutics, Takeda, Teladoc Health, TiGenix, UCB Inc., Vial, Vifor; reports lecture fees from Abbvie, Amgen, Ferring Pharmaceuticals Inc., Gilead, Janssen, Mylan, Pfizer, Takeda.
FM: served as speaker and received honoraria from: Abbvie, Arena, Biogen, Bristol-Myers Squibb, Falk, Ferring, Hospira, Janssen, Laboratórios Vitoria, Pfizer, Lilly, Merck Sharp & Dohme, Sandoz, Takeda, UCB, Vifor.
SP, SB & LH: employee, paid consultant and/or shareholder of MRM Health.

Introduction

Microplastic particles (MPs), defined as plastic fragments smaller than 5 mm, are pervasive pollutants that accumulate in ecosystems and the human food chain (1). Literature points at various health risks, including the induction of carcinogenesis (2). Emerging evidence from animal models (3) suggests that MPs may influence the gut microbiome, but the impact on the human gut microbiome remains poorly understood.

Aims & Methods

This study aimed to evaluate how various MP types influence gut microbial composition and metabolism using an ex vivo bioreactor model, with a focus on exploring potential carcinogenic effects arising from MP–microbiome interactions.
Stool samples from healthy donors were used to inoculate bioreactor cultures, which were maintained under anaerobic conditions for five days with daily nutrient feeding. These cultures were exposed to five common MP types—polystyrene (PS), polypropylene (PP), low-density polyethylene (LDPE), poly(methyl methacrylate) (PMMA), and polyethylene terephthalate (PET)—at concentrations mimicking both estimated human exposure (4) and higher doses to assess dose-dependence. Cell viability and total bacterial counts, as well as culture pH, were measured throughout the experiment. To further explore potential microbiome and metabolic alterations, 16S rRNA gene sequencing and targeted metabolomics were performed.

Results

MP exposure did not lead to significant changes in total or viable bacterial cell counts. However, MP-treated cultures exhibited a consistent and significant decrease in pH compared to controls, suggesting changes in microbial metabolic activity. Microbiome sequencing revealed plastic-type-dependent alterations in microbial composition, with certain bacterial taxa increasing or decreasing in abundance depending on the MP type. These shifts occurred across multiple genera within diverse families, including Lachnospiraceae, Oscillospiraceae, Enterobacteriaceae, and Ruminococcaceae, with the majority of changes occurring withing the phylum Bacillota. These compositional changes were accompanied by shifts in the metabolomic profiles, some of which correlated with the observed pH changes. Several plastic types induced changes in valeric acid levels, while individual MP types were associated with alterations in distinct metabolites such as uracil, lactic acid, and acetic acid.

Conclusion

This study demonstrates that MPs can affect gut microbial activity and metabolism without significantly altering overall bacterial abundance in the short term. The variation in microbiome response across plastic types highlights the complexity of MP-microbiome interactions. Changes in faecal pH are known to be associated with various gastrointestinal diseases (5). Interestingly, some of the MP-induced changes in microbial composition resembled patterns linked to diseases such as depression and colorectal cancer. In contrast, other changes showed patterns that differed from those typically associated with conditions like Parkinson’s disease and irritable bowel syndrome, highlighting the complexity of these associations and the dependency on the context. The bioreactor model provided a controlled environment for identifying direct MP-microbiome interactions; however, in humans, additional factors like diet, immune response, and individual microbiome variations play a role in determining long-term effects. Exposure duration may also play a critical role in shaping the extent and nature of microbiome alterations (6), underscoring the need for further research into the effects of chronic MP exposure and associated health risks.

References

(1) Mamun, A. A., Prasetya, T. A. E., Dewi, I. R., & Ahmad, M. (2023). Microplastics in human food chains: Food becoming a threat to health safety. The Science of the total environment, 858(Pt 1), 159834. https://doi.org/10.1016/j.scitotenv.2022.159834
(2) Kumar, R., Manna, C., Padha, S., Verma, A., Sharma, P., Dhar, A., Ghosh, A., & Bhattacharya, P. (2022). Micro(nano)plastics pollution and human health: How plastics can induce carcinogenesis to humans?. Chemosphere, 298, 134267. https://doi.org/10.1016/j.chemosphere.2022.134267
(3) Hirt, N., & Body-Malapel, M. (2020). Immunotoxicity and intestinal effects of nano- and microplastics: a review of the literature. Particle and fibre toxicology, 17(1), 57. https://doi.org/10.1186/s12989-020-00387-7
(4) Senathirajah, K., Attwood, S., Bhagwat, G., Carbery, M., Wilson, S., & Palanisami, T. (2021). Estimation of the mass of microplastics ingested - A pivotal first step towards human health risk assessment. Journal of hazardous materials, 404(Pt B), 124004. https://doi.org/10.1016/j.jhazmat.2020.124004
(5) Yamamura, R., Inoue, K. Y., Nishino, K., & Yamasaki, S. (2023). Intestinal and fecal pH in human health. Frontiers in Microbiomes, 2, 1192316.
(6) Yin, L., Yang, M., Teng, A., Ni, C., Wang, P., & Tang, S. (2025). Unraveling Microplastic Effects on Gut Microbiota across Various Animals Using Machine Learning. ACS nano, 19(1), 369–380. https://doi.org/10.1021/acsnano.4c07885

Introduction

Celiac disease (CeD) is triggered by gluten peptides that bind to HLA-DQ2 or -DQ8 in the small intestine leading to gluten specific Th1 T cell expansion and villous atrophy. The oral transglutaminase 2 inhibitor Zed1227 blocks the deamidation of gliadin peptides, a prerequisite for effective gliadin antigen presentation. The CEC-3 clinical trial demonstrated that Zed1227 effectively prevented mucosal damage in 160 CeD patients in remission challenged with 3 g gluten daily for 6 weeks (Schuppan D et al, NEJM 2021).

Aims & Methods

We analyzed regulatory T cell (Treg) gene expression pre and post gluten challenge in the 100 mg/d Zed1227 vs placebo treated patients using RNAseq from the patients’ small intestinal biopsies (Dotsenko V et al, Nat Immunol 2024). We focused on genes of Treg subtypes and related cells. Subanalysis was performed via different gene set enrichments for Treg subsets, regulatory pathways and cell types. Genes that were enhanced in favorable vs unfavorable villous height to crypt depth (V/C) ratio outcomes, also after placebo treatment, were analyzed. We further analyzed pre-disposing genes before challenge, favoring a beneficial or negative post challenge V/C outcome.

Results

93% of the top 100 most significant regulatory genes were upregulated in Zed1227-treated patients compared to those treated with a placebo. These regulatory genes included TGF-beta signaling genes such as Smad7 and MERTK, cholesterol- and fatty acid related genes, such as ACAT1 and NPC1L1, specific autophagy/apoptosis related genes such as VPS35, SQSTM1, BMF, and inhibitory genes like SOCS2. Zed1227 treated patient samples with a maintained post challenge V/C ratio showed evidence for upregulated iTreg genes. Interestingly, Treg genes from placebo-treated subjects were upregulated in correlation with better inflammatory control under gluten challenge compared to worse control.
Moreover, in pre-challenge duodenal samples we identified protective genes that help maintain tolerance during gluten challenge, as reflected by a better post challenge V/C ratio. Examples are ubiquitin ligase RNF167, BAD, FOXN3, BAG1 and CLAUDIN15, whereas other genes like IRF9, CD96 and HLA-/TAP1 were predictive of a more pronounced V/C ratio decline after gluten exposure.

Conclusion

Our data derived from superbly analyzed pre- and post challenge duodenal biopsies demonstrate an active anti-inflammatory immune modulation, largely Treg cell mediated, in CeD in patients prior to and during gluten challenge. Zed1227 treatment potently activates this regulatory response. The balance of several beneficial vs detrimental predisposing Treg-related genes may play a causative role in the variable sensitivity of CeD patients to gluten exposure. Our data support future Treg based therapies to attenuate inflammation in CeD and other (intestinal) autoimmune diseases.

References

Schuppan, D., Mäki, M., Lundin, K. E. A., Isola, J., Friesing-Sosnik, T., Taavela, J., Popp, A., Koskenpato, J., Langhorst, J., Hovde, Ø., Lähdeaho, M. L., Fusco, S., Schumann, M., Török, H. P., Kupcinskas, J., Zopf, Y., Lohse, A. W., Scheinin, M., Kull, K., Biedermann, L., … CEC-3 Trial Group (2021). A Randomized Trial of a Transglutaminase 2 Inhibitor for Celiac Disease. The New England journal of medicine, 385(1), 35–45. https://doi.org/10.1056/NEJMoa2032441
Dotsenko, V., Tewes, B., Hils, M., Pasternack, R., Isola, J., Taavela, J., Popp, A., Sarin, J., Huhtala, H., Hiltunen, P., Zimmermann, T., Mohrbacher, R., Greinwald, R., Lundin, K. E. A., Schuppan, D., Mäki, M., Viiri, K., & CEC-3 Investigators (2024). Transcriptomic analysis of intestine following administration of a transglutaminase 2 inhibitor to prevent gluten-induced intestinal damage in celiac disease. Nature immunology, 25(7), 1218–1230. https://doi.org/10.1038/s41590-024-01867-0

Disclosure

V.D. and K.V. received funding from Dr. Falk Pharma to Tampere University to conduct the study. B.T., T.Z., R.M. and R.G. are employees of Dr. Falk Pharma. The data presented here are the subject of patent applications EP24173619.8 and EP24173615.6 filed by Dr. Falk Pharma, and B.T., T.Z., R.M., R.G., V.D. and K.V. are inventors on these applications. M.H. and R.P. are employees of Zedira. J.T. is a consultant for Jilab Oy and Dr. Falk Pharma. K.E.A.L. is a consultant for Amyra, Bioniz Pharmaceuticals, Chugai Pharmaceutical, Dr. Falk Pharma, Itrexon Actobios, TOPAS Therapeutics and Takeda California. D.S. is the data and safety monitor for Boehringer Ingelheim (Phil.) and is a consultant for the Dr. Falk Pharma, Takeda, Immunic, Sanofi and TOPAS Therapeutics. J.I. is the owner of Jilab Oy. M.M. is the founder, owner and Chair of the Board of Maki HealthTech (MHT). MHT receives Management/Advisory Affiliation fees from Dr. Falk Pharma and other funding not related to the research from Topas Therapeutics, Calypso Biotech, Vaccitech, ImmunogenX, Equillium and Immunic. MHT holds patents (patent number 7361480 (United States) and European Patent Office Number 1390753) licensed to Labsystems Diagnostics from where MHT receives royalties via Tampere University Hospital. All other authors declare no competing interests.

Introduction

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a chronic liver disease estimated to affect 38% of the global adult population. Activation of hepatic progenitor cells (HPCs) and concurrent proliferation of reactive bile ducts (collectively referred to as ductular reaction) have been associated with advanced MASLD, liver fibrosis, and damage, yet their exact role during disease progression remains unclear.

Aims & Methods

This study aimed to investigate the prognostic and biological relevance of ductular reaction in the progression of MASLD by using a multi-omics approach. In this study, a total of 251 patient samples were included. Spatially resolved multi-omics techniques were implemented to characterise the HPC niche in advanced MASLD using laser microdissection combined with high-throughput RNA sequencing (RNA-seq), nanoString CosMx molecular profiling, followed by Lunaphore COMET proteome multiplexing at single-cell resolution. Cell deconvolution by integrating single nucleus RNA-seq of MASH biopsy samples was implemented to determine cell of origin. Bulk RNA-seq analysis of 206 MASLD biopsy samples, stratified by HPC gene signatures, was used to examine differences in clinical parameters and genotype. Finally, clinico-pathological prognostic features of HPCs were assessed in baseline biopsies of MASLD patients with a follow-up biopsy at least one year apart.

Results

A high-throughput transcriptomics analysis of the HPC niche identified 3,282 differentially expressed genes enriched in pro-inflammatory cytokines (e.g. CXCL6, CCL19, CCL2, CCL5, CCL21, SPP1), inflammatory cell markers (e.g. CCR2, CD48, CD8a, CD96, CXCR3, S100A4), as well as antigen presenting genes. Spatial transcriptomics combined with integrated single nucleus RNA-seq uncovered distinct subsets of ductular reaction based on the expression of inflammatory markers SPP1+ and CCL2+. Notably, SPP1+CCL2- ductular reaction localised predominantly within the parenchyma and peri-portal region, while CCL2+SPP1- ductular reaction was confined mainly to the portal tract. Neighbourhood analysis highlighted antigen-presenting cells and IGHA1+IGHG1+ plasma cells in close proximity to ductular reaction. Spatial proteomics validation identified CD48 and HLA-DR-expressing macrophages as key immune cells of the HPC niche. Furthermore, the resulting HPC gene signature stratified transcriptomics data from 206 NAFLD biopsies into distinct clusters characterised by presence of steatohepatitis, high disease activity, fibrosis and AST levels. In a dual biopsy cohort, HPC presence in the parenchyma at baseline was significantly associated with progression of at least 1 fibrosis stage and was able to predict fibrosis progression in MASLD in a binary logistic regression model.

Conclusion

This study identified a pro-inflammatory role for HPCs in patients with MASLD. Furthermore, we showed the clinical relevance of using HPC signatures associated with disease activity and inflammation, as well as their ability to predict fibrosis progression in MASLD patients.