Ex Vivo Comparison Of Combination Versus Single Therapeutic Pathway Inhibition In Human Intestinal Mucosa From Patients With Inflammatory Bowel Disease

NCT07824726 | Not Yet Recruiting

• 1 other identifier: COMBI-IBD
• Study Type: interventional
• Target: 30
• Locations: 1 country
• Sites: 1

Brief Summary

The purpose of this study is to evaluate whether simultaneously targeting three inflammatory pathways (TL1A, IL-23, and α4β7) produces a stronger anti-inflammatory effect in intestinal tissue from patients with Inflammatory Bowel Disease (IBD) compared with targeting individual pathways. The study aims to investigate whether this combined approach may help overcome limitations associated with single-pathway inhibition. This is a single-center, interventional ex vivo study conducted at IRCCS Ospedale San Raffaele. The study includes 30 participants divided into three cohorts: 10 patients with Ulcerative Colitis, 10 patients with Crohn's Disease, and 10 non-IBD control participants. Participant involvement is limited to a single day and is integrated into a clinically indicated, routinely scheduled colonoscopy and routine phlebotomy. During the scheduled colonoscopy, 8 additional mucosal biopsies are collected for research purposes, together with an additional 2 mL blood sample. No additional endoscopic procedure or separate study visit is required. No investigational drug or treatment is administered directly to participants. The collected intestinal biopsies are instead treated ex vivo in the laboratory with ATTO-1091, individual pathway inhibitors, or control conditions for 16 hours. Biological and molecular responses are evaluated using laboratory techniques including ELISA and RNA sequencing to assess inflammatory markers and gene expression. Laboratory personnel performing molecular and transcriptomic analyses are blinded to participants' clinical profiles until final data analysis.

Conditions

IBD (Inflammatory Bowel Disease); Ulcerative Colitis (UC); Crohn Disease (CD)

Keywords

IBD; Ulcerative Colitis; Crohn's Disease; Combination Therapy; Ex Vivo Human Organ Culture

Outcome Measures

Primary Outcomes (3)

• Concentration of pro-inflammatory cytokines in intestinal biopsy culture supernatants following ex vivo treatment with ATTO-1091

  Pro-inflammatory cytokine concentrations will be quantified by enzyme-linked immunosorbent assay (ELISA) in culture supernatants collected after 16 hours of ex vivo incubation of intestinal biopsies with ATTO-1091, individual pathway inhibitors, or the corresponding control condition. Cytokine concentrations following ATTO-1091 exposure will be compared with those measured in control cultures and in cultures exposed to individual pathway inhibitors to quantify the anti-inflammatory effect of simultaneous TL1A, IL-23, and α4β7 inhibition. Each selected cytokine will be analyzed separately and reported as its concentration in the culture supernatant using the concentration unit specified for the corresponding ELISA assay. Comparisons between experimental conditions will be performed using Student's t-test or analysis of variance (ANOVA), as appropriate, with correction for multiple comparisons.

  16 hours after ex vivo exposure

• Change in gene expression following ex vivo treatment with ATTO-1091, expressed as log2 fold change

  Gene expression profiles will be assessed by RNA sequencing (RNA-seq) in intestinal biopsies following 16 hours of ex vivo incubation with ATTO-1091 or the corresponding comparator conditions. Sequencing will be performed using paired-end reads (2 × 150 bp), targeting approximately 30 million read pairs per sample. After quality control, trimming, alignment to the human reference genome, and generation of gene-level counts, differential gene expression will be analyzed using DESeq2. Gene expression profiles following ATTO-1091 exposure will be compared with the corresponding control and single-pathway inhibitor conditions to identify genes significantly modulated by simultaneous TL1A, IL-23, and α4β7 inhibition. Changes in gene expression between experimental conditions will be expressed as log2 fold change (log2FC), with statistical significance assessed using adjusted P values.

  After 16 hours of ex vivo incubation

• Enrichment of disease-associated inflammatory biological pathways following ex vivo treatment with ATTO-1091

  Functional and pathway enrichment analyses will be performed using RNA-seq-derived differential gene expression data from intestinal mucosal tissue following 16 hours of ex vivo exposure to ATTO-1091, individual pathway inhibitors, or control conditions. Pathway enrichment will be assessed from differential gene expression data to identify disease-associated inflammatory biological processes and pathways modulated by simultaneous TL1A, IL-23, and α4β7 inhibition. Pathway enrichment following ATTO-1091 exposure will be compared with that observed under control conditions and following individual pathway inhibition. Biological processes and pathways with an adjusted P value ≤0.05 will be considered significantly enriched. The enrichment measure is an analytical output of pathway enrichment analysis and is not a clinical scale with predefined minimum or maximum values.

  After 16 hours of ex vivo incubation

Secondary Outcomes (4)

• Correlation between ex vivo gene expression response to ATTO-1091 and disease duration in participants with IBD

  Through study completion, up to 12 months

• Correlation between ex vivo gene expression response to ATTO-1091 and Total Mayo Score in participants with Ulcerative Colitis

  Through study completion, up to 12 months

• Correlation between ex vivo gene expression response to ATTO-1091 and Crohn's Disease Activity Index in participants with Crohn's Disease

  Through study completion, up to 12 months

• Correlation between ex vivo gene expression response to ATTO-1091 and participant age

  Through study completion, up to 12 months

Study Arms (3)

Control group - NO IBD

OTHER

Non-IBD control cohort consisting of individuals without inflammatory bowel disease undergoing scheduled standard-of-care colonoscopy. During the procedure, 8 additional mucosal biopsies and 2 mL of blood are collected. Collected mucosal tissue is cultured ex vivo for 16 hours with the tri-specific inhibitor ATTO-1091, individual monotherapies (tulisokibart, risankizumab, vedolizumab), or vehicle control to evaluate baseline non-inflamed mucosal responses.

Procedure: Collection of 8 additional intestinal mucosal biopsies during standard-of-care endoscopy for subsequent ex vivo testing; Procedure: Collection of an additional 2 mL blood sample

Crohn's Disease (CD)

OTHER

Crohn's Disease (CD) cohort consisting of individuals with confirmed CD undergoing scheduled standard-of-care colonoscopy. During the procedure, 8 additional mucosal biopsies and 2 mL of blood are collected. Collected mucosal tissue is cultured ex vivo for 16 hours with the tri-specific inhibitor ATTO-1091, individual monotherapies (tulisokibart, risankizumab, vedolizumab), or vehicle control to evaluate mucosal responses in CD tissue.

Procedure: Collection of 8 additional intestinal mucosal biopsies during standard-of-care endoscopy for subsequent ex vivo testing; Procedure: Collection of an additional 2 mL blood sample

Ulcerative Colitis (UC)

OTHER

Ulcerative Colitis (UC) cohort consisting of individuals with confirmed UC undergoing scheduled standard-of-care colonoscopy. During the procedure, 8 additional mucosal biopsies and 2 mL of blood are collected. Collected mucosal tissue is cultured ex vivo for 16 hours with the tri-specific inhibitor ATTO-1091, individual monotherapies (tulisokibart, risankizumab, vedolizumab), or vehicle control to evaluate mucosal responses in UC tissue.

Procedure: Collection of 8 additional intestinal mucosal biopsies during standard-of-care endoscopy for subsequent ex vivo testing; Procedure: Collection of an additional 2 mL blood sample

Interventions

Collection of 8 additional intestinal mucosal biopsies during standard-of-care endoscopy for subsequent ex vivo testing PROCEDURE

Eight additional mucosal biopsies will be collected during a clinically indicated, scheduled standard-of-care colonoscopy. Biopsies will be obtained according to a standardized sampling protocol, with the anatomical site of each biopsy documented. The additional biopsies are collected for research purposes and subsequently processed for ex vivo laboratory analyses. No investigational product is administered to study participants.

Control group - NO IBD; Crohn's Disease (CD); Ulcerative Colitis (UC)

Collection of an additional 2 mL blood sample PROCEDURE

An additional 2 mL blood sample will be collected during routine phlebotomy performed in conjunction with the scheduled study visit. The additional blood collection is performed for research purposes and does not require a separate study visit.

Control group - NO IBD; Crohn's Disease (CD); Ulcerative Colitis (UC)

Eligibility Criteria

• Age: 18 Years+
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• Patients of at least 18 years of age
• Able to comply with the study procedures and to sign an informed consent form
• Established diagnosis of UC or CD, with indication to start any biological or small molecule agents as per standard of care:
• UC patients with a clinical indication to start biologics or small molecules as per standard of care, with a Total Mayo Score of 6-12 (moderate-severe disease) and an endoscopic subscore ≥ 2; CD patients with a clinical indication to start biologics or small molecules as per standard of care, with a clinical CDAI score of 220-600 (moderate-severe disease) and endoscopic evidence of active mucosal inflammation/ulceration upon baseline examination;
• \- Patients with IBS or individuals undergoing colonoscopy for CRC prevention or routine surveillance, with no prior diagnosis of inflammatory bowel disease and no endoscopic evidence of intestinal mucosal inflammation.

You may not qualify if:

• Absolute contraindications to colonoscopy procedures
• UC or CD patients in endoscopic remission
• IBS or patients undergoing CRC prevention surveillance with inflamed mucosa during the endoscopy
• Patients of at least 18 years of age unable to comply with the study procedures and to sign an informed consent form
• Pregnancy or breastfeeding

Sponsors & Collaborators

• PhoenixLAB srls: lead
• IRCCS San Raffaele: collaborator

Study Sites (1)

IRCCS San Raffaele

Milan, MI, 20131, Italy

Location

Related Publications (3)

• Al-Lamki RS, Bradley JR, Pober JS. Human Organ Culture: Updating the Approach to Bridge the Gap from In Vitro to In Vivo in Inflammation, Cancer, and Stem Cell Biology. Front Med (Lausanne). 2017 Sep 11;4:148. doi: 10.3389/fmed.2017.00148. eCollection 2017.

  PMID: 28955710 BACKGROUND

• Feuerstein JD, Ho EY, Shmidt E, Singh H, Falck-Ytter Y, Sultan S, Terdiman JP; American Gastroenterological Association Institute Clinical Guidelines Committee. AGA Clinical Practice Guidelines on the Medical Management of Moderate to Severe Luminal and Perianal Fistulizing Crohn's Disease. Gastroenterology. 2021 Jun;160(7):2496-2508. doi: 10.1053/j.gastro.2021.04.022. No abstract available.

  PMID: 34051983 BACKGROUND

• Feuerstein JD, Isaacs KL, Schneider Y, Siddique SM, Falck-Ytter Y, Singh S; AGA Institute Clinical Guidelines Committee. AGA Clinical Practice Guidelines on the Management of Moderate to Severe Ulcerative Colitis. Gastroenterology. 2020 Apr;158(5):1450-1461. doi: 10.1053/j.gastro.2020.01.006. Epub 2020 Jan 13. No abstract available.

  PMID: 31945371 BACKGROUND

MeSH Terms

Conditions

Colitis, Ulcerative; Inflammatory Bowel Diseases; Crohn Disease

Interventions

Blood Specimen Collection

Condition Hierarchy (Ancestors)

Colitis; Gastroenteritis; Gastrointestinal Diseases; Digestive System Diseases; Colonic Diseases; Intestinal Diseases

Intervention Hierarchy (Ancestors)

Specimen Handling; Clinical Laboratory Techniques; Diagnostic Techniques and Procedures; Diagnosis; Punctures; Surgical Procedures, Operative; Investigative Techniques

Study Officials

• Silvio Danese, MD, PhD

  IRCCS San Raffaele

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Silvia D'Alessio, PhD

CONTACT

+393394383067; silviadalessio@phoenixlab1.com

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: NON RANDOMIZED
• Masking: NONE
• Masking Details: This is an open-label study. Experimental compounds and vehicle controls are applied strictly ex vivo to mucosal biopsy tissue, with no in vivo intervention or masking required for participants or care providers. Laboratory personnel performing molecular and transcriptomic analyses are blinded to participants' clinical profiles until final data analysis.
• Purpose: BASIC SCIENCE
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Model Details: This is an interventional ex vivo study using organotypic mucosal tissue cultures. Participants are enrolled into three parallel diagnostic cohorts based on their pre-existing clinical diagnosis: Ulcerative Colitis (UC, n=10), Crohn's Disease (CD, n=10), and non-IBD controls (n=10). During a scheduled standard-of-care colonoscopy, 8 additional mucosal biopsies are collected from each participant, together with an additional 2 mL blood sample during routine phlebotomy. Collected mucosal tissue is cultured ex vivo for 16 hours and exposed under standardized laboratory conditions to the experimental compound ATTO-1091, which simultaneously inhibits the TL1A, IL-23, and α4β7 pathways, individual pathway inhibitors (tulisokibart, risankizumab, and vedolizumab), or control conditions. No investigational product is administered directly to participants; all pharmacological exposures occur exclusively ex vivo on the collected tissue.

Study Record Dates

• First Submitted: September 4, 2026
• First Posted: September 17, 2026
• Study Start: October 1, 2026
• Primary Completion: October 1, 2026
• Study Completion: October 1, 2026
• Last Updated: September 17, 2026

Record last verified: 2026-09

Locations

IT (1)