The Microbial Impact on Intestinal Fibrosis and the Associated Immune Microenvironment in Crohn's Disease

NCT06720961 | Not Yet Recruiting

• 2 other identifiers: PRIN-2022RKE4LBando MUR 2022 PRIN-2022RKE4L
• Study Type: observational
• Target: 20
• Locations: 1 country
• Sites: 1

Brief Summary

The goal of this study is to find out if there is a direct connection between an imbalance of gut bacteria and the development of scar tissue in the gut by identifying important bacterial proteins found in scarred gut tissue. Our aim is to identify which types of cells and biological processes are affected by these bacterial proteins in people with Crohn's Disease. We will also study how these bacterial proteins cause changes in 3D models of gut fibrosis.

Conditions

Crohn Disease (CD)

Keywords

Crohn Disease; Fibrosis; Dysbiosis; Microbiota; Fibrostenosis

Outcome Measures

Primary Outcomes (1)

• To identify the cellular subtypes and molecular pathways impacted by the specific bacterial factors during CD-associated fibrosis.

  Control subjects' and CD-derived surgical specimens will be processed to obtain a cell suspension, that will be frozen and stored for the subsequent cell sorting. Frozen CD and healthy cell suspensions will be then thawed and undergo FACS for specific cell markers (CD31 for endothelial cells, EpCam for epithelial cells, CD90 for fibroblasts, CD45 for leukocytes, including CD4 and CD8 for T cells, CD20 for B cells, CD14 and CD163 for macrophages (MΦ), CD11b and c for dendritic cells (DCs). Single-cell populations will undergo library preparation and will be analyzed by ribo-minus RNAseq at 30M reads of depth. Metatranscriptomics for profiling the microbial composition, as well as the transcriptomics to determine both the differential gene expression (DGE) and the Gene Set Enrichment Analysis (GSEA), will be performed.

  Experiments and analysis: 11th to 36th month

Secondary Outcomes (1)

• To unravel the cellular and molecular mechanisms and dynamics induced by the bacterial factors in 3D models of intestinal fibrosis

  Experiments and analysis: 11th to 36th month

Other Outcomes (1)

• To validate in vitro results in an animal model by assessing the role of microbial factors in inducing fibrosis in an entire organism. Moreover, we will provide a possible therapeutic approach with strong clinical implications in the future

  Experiments on in-vivo models: 13th to 24th month

Study Arms (2)

CD Patients

CD patients with established diagnosis and additionally classified according to 3 stages: B1 (more inflammatory, non-stricturing), B2 (stricturing, non-penetrating) and B3 (stricturing and penetrating). Patients will be stratified on the basis of CT, MRI or ultrasound analysis (stratification already known before surgery)

Procedure: Collection of surgical specimen; Procedure: Collection of stool sample

Non-IBD Patients

Patients with other pathologies but not affected by IBD according to the previously reported clinical and endoscopic evaluation criteria (ex. diverticulitis)

Procedure: Collection of surgical specimen; Procedure: Collection of stool sample

Interventions

Collection of surgical specimen PROCEDURE

Specimens of CD patients and non-IBD related patients will be collected during the surgery performed for clinical practice, without other risks for the patients, since we will use only leftover material after pathologist analysis. All of the collected surgical samples will be used for this project and, for this reason, they will not be conserved after the research period. Any residual samples will be destroyed.

CD Patients; Non-IBD Patients

Collection of stool sample PROCEDURE

One week before the surgery to remove the stricture, fecal samples from CD patients belonging to B1, B2, B3 groups will be collected. They will be used for the exploratory objective. All of the collected fecal samples will be used for this project and, for this reason, they will not be conserved after the research period. Any residual samples will be destroyed.

CD Patients; Non-IBD Patients

Eligibility Criteria

• Age: 18 Years - 69 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Non-Probability Sample

Study Population

We will enroll 15 CD patients and 5 patients without IBD-related diseases (ex. diverticulitis) undergoing surgery according to the normal clinical practice for a total of 20 patients as following: \- 15 patients with CD (5 patients/stage: B1 (more inflammatory, non-structuring), B2 (structuring, non-penetrating) and B3 (structuring and penetrating). Patients will be stratified according to CT, MRI or ultrasound analysis (stratification already known before surgery) \- 5 patients without IBD-related diseases (e.g. diverticulitis) All patients will be recruited only from San Raffaele Hospital (OSR). The participation is voluntary and the patient is allowed to refuse further participation in the protocol whenever he/she wants. Participation in the study will not change the diagnostic-therapeutic treatment that patients undergo following the clinical practice, since patients will continue to be treated in accordance with clinical practice regardless of the results of this study.

You may qualify if:

• Adult patients ≥18 and \<70 years
• Patients able to autonomously sign the informed consent
• Non pregnant or breastfeeding patients
• For CD patients: CD patients with established diagnosis and additionally classified according to 3 stages: B1 (more inflammatory, non-stricturing), B2 (stricturing, non-penetrating) and B3 (stricturing and penetrating). Patients will be stratified on the basis of CT, MRI or ultrasound analysis (stratification already known before surgery)
• For non-IBD patients: patients with other pathologies but not affected by IBD according to the previously reported clinical and endoscopic evaluation criteria (ex. diverticulitis)

You may not qualify if:

• Patients \<18 years or \> 70 years
• Pregnant or breastfeeding patients
• For CD Patients: Subjects with CD who do not meet evaluation criteria described above
• For non-IBD patients: Patients undergoing anti-inflammatory and/or immunosuppressive treatments for IBD-related diseases

Sponsors & Collaborators

• IRCCS San Raffaele: lead
• University of Florence: collaborator
• Humanitas Research Hospital IRCCS, Rozzano-Milan: collaborator
• Ministero dell'Istruzione, dell'Università e della Ricerca: collaborator
• Università Vita-Salute San Raffaele (MI): collaborator

Study Sites (1)

IRCCS San Raffaele

Milan, Italy, 20132, Italy

Location

Biospecimen

Retention: SAMPLES WITH DNA

Bioptic surgical specimens will be collected according to standard of care and, for the study, we will use leftover material after pathologist analysis. One week before the surgery the collection of fecal material from CD and from non-IBD patients will be performed ad hoc for clinical research purposes of this study.

MeSH Terms

Conditions

Crohn Disease; Fibrosis; Dysbiosis

Condition Hierarchy (Ancestors)

Inflammatory Bowel Diseases; Gastroenteritis; Gastrointestinal Diseases; Digestive System Diseases; Intestinal Diseases; Pathologic Processes; Pathological Conditions, Signs and Symptoms

Study Officials

• Silvio Danese, PhD-MD

  IRCCS San Raffaele

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Silvio Danese, PhD.-MD

CONTACT

+39 0226432807; danese.silvio@hsr.it

Federica Ungaro, PhD.

CONTACT

+39 0226437864; ungaro.federica@hsr.it

Study Design

• Study Type: observational
• Observational Model: CASE CONTROL
• Time Perspective: CROSS SECTIONAL
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Professor

Study Record Dates

• First Submitted: December 3, 2024
• First Posted: December 6, 2024
• Study Start: April 1, 2025
• Primary Completion: January 1, 2026
• Study Completion (Estimated): January 1, 2027
• Last Updated: February 14, 2025

Record last verified: 2025-02

Locations

IT (1)