Ex Vivo Comparison Of Combination Versus Single Therapeutic Pathway Inhibition In Human Intestinal Mucosa From Patients With Inflammatory Bowel Disease
试验速览
- 阶段
- 不适用
- 状态
- 尚未招募
- 发起方
- 入组人数
- 30
- 试验地点
- 1
- 主要终点
- Concentration of pro-inflammatory cytokines in intestinal biopsy culture supernatants following ex vivo treatment with ATTO-1091
研究概览
简要总结
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.
详细描述
The study is built on the hypothesis that the simultaneous blockade of TL1A, IL-23, and α4β7 pathways will exert a synergistic anti-inflammatory effect on the intestinal mucosa of IBD patients compared to single-agent inhibition, thereby potentially overcoming resistance to standard biological therapies.
To test this, a monocentric, national, interventional ex vivo clinical study will be conducted at the IRCCS Ospedale San Raffaele using organotypic tissue cultures.
The parallel-group design is based on three distinct clinical cohorts consisting of participants with Ulcerative Colitis, participants with Crohn's Disease, and non-IBD control subjects.
To minimize bias and ensure scientific integrity, laboratory operators performing the molecular and transcriptomic analyses will be blinded to the clinical and response data of the source tissue until final statistical analysis.
Participants will undergo a clinically indicated, routinely scheduled standard-of-care colonoscopy. During the procedure, 8 additional mucosal biopsies will be collected for research purposes according to a standardized sampling protocol, together with an additional 2 mL blood sample collected during routine phlebotomy. No investigational product or treatment will be administered directly to study participants. Instead, the active experimental phase will be conducted entirely ex vivo in the laboratory, where the collected intestinal tissue samples will be exposed to ATTO-1091, individual pathway inhibitors, or control conditions for a standardized culture period of 16 hours.
研究设计
- 研究类型
- Interventional
- 分配方式
- Non Randomized
- 干预模型
- Parallel
- 主要目的
- Basic Science
- 盲法
- None
盲法说明
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.
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •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.
排除标准
- •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
研究组 & 干预措施
Control group - NO IBD
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.
干预措施: Collection of 8 additional intestinal mucosal biopsies during standard-of-care endoscopy for subsequent ex vivo testing (Procedure)
Control group - NO IBD
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.
干预措施: Collection of an additional 2 mL blood sample (Procedure)
Crohn's Disease (CD)
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.
干预措施: Collection of 8 additional intestinal mucosal biopsies during standard-of-care endoscopy for subsequent ex vivo testing (Procedure)
Crohn's Disease (CD)
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.
干预措施: Collection of an additional 2 mL blood sample (Procedure)
Ulcerative Colitis (UC)
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.
干预措施: Collection of 8 additional intestinal mucosal biopsies during standard-of-care endoscopy for subsequent ex vivo testing (Procedure)
Ulcerative Colitis (UC)
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.
干预措施: Collection of an additional 2 mL blood sample (Procedure)
结局指标
主要结局
Concentration of pro-inflammatory cytokines in intestinal biopsy culture supernatants following ex vivo treatment with ATTO-1091
时间窗: 16 hours after ex vivo exposure
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.
Change in gene expression following ex vivo treatment with ATTO-1091, expressed as log2 fold change
时间窗: After 16 hours of ex vivo incubation
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.
Enrichment of disease-associated inflammatory biological pathways following ex vivo treatment with ATTO-1091
时间窗: After 16 hours of ex vivo incubation
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.
次要结局
- 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)