Single-cell Study of the Systemic Immune Response to Controlled Gluten Intake in Pediatric Celiac Disease
试验速览
- 阶段
- 不适用
- 状态
- 招募中
- 发起方
- 入组人数
- 45
- 试验地点
- 1
- 主要终点
- Change in peripheral blood immune cell gene expression after controlled gluten ingestion
研究概览
简要总结
This study investigates how the immune system of children with celiac disease responds to controlled, small amounts of gluten. Children on a strict gluten-free diet are randomly assigned to receive either placebo, 50 mg of gluten, or 5 g of gluten once daily for three days, simulating real-life accidental exposure or dietary transgression. Blood samples are collected on Day 1 (before gluten intake) and Day 8 (five days after the last dose). Stool and urine samples are also collected for complementary analyses.
Using single-cell ribonucleic acid (RNA) sequencing, T-cell receptor sequencing, microRNA profiling, and exploratory metabolomics, the study aims to characterize changes in immune cell populations and gene expression after gluten exposure. The objective is to determine whether even very small amounts of gluten induce measurable systemic immune responses and whether these responses differ according to the dose administered. Understanding these mechanisms may support the development of new biomarkers and improve clinical management of pediatric celiac disease.
详细描述
Celiac disease is a chronic, immune-mediated enteropathy triggered by gluten ingestion in genetically predisposed individuals. In pediatric patients, strict adherence to a gluten-free diet (GFD) leads to mucosal recovery and clinical improvement; however, the systemic immunological landscape during long-term GFD remains incompletely understood. Accidental gluten exposure is common in daily life and may trigger subclinical immune activation even in the absence of overt symptoms. The biological consequences of such low-dose exposures, particularly at the single-cell level, are not fully characterized in children.
CELLiomicS (Single-cell Immune Response to Controlled Gluten Ingestion in Pediatric Celiac Disease) is a randomized, controlled, parallel-group clinical study designed to characterize systemic immune responses to controlled gluten ingestion in pediatric patients with established celiac disease. Eligible participants are children aged 8-14 years who have followed a strict GFD for at least 18 months before enrollment. The study plans to recruit approximately 45 participants with biopsy- or serology-confirmed celiac disease.
Participants are randomized into three intervention groups: placebo, 50 mg gluten, or 5 g gluten. The selected gluten doses represent two clinically relevant scenarios: (1) accidental low-level exposure (50 mg), and (2) dietary transgression (5 g), both below the threshold typically required to cause serological relapse. Gluten or placebo is administered once daily on Days 1, 2, and 3. Blood samples are obtained on Day 1 (baseline) and Day 8 (five days after the final dose). Stool and urine samples are also collected at the same time points for complementary analyses, including fecal gluten immunogenic peptides (GIP) and exploratory metabolomics.
Peripheral blood mononuclear cells are processed for high-dimensional immune profiling using single-cell RNA sequencing and T-cell receptor (TCR) sequencing (10x Genomics). These analyses enable characterization of immune cell composition, transcriptional activity, and clonal T-cell dynamics before and after gluten challenge. In addition, exosomal microRNA expression is assessed as approved by the ethics committee amendment SICEIA-2025-001434, and untargeted metabolomic profiling of blood, stool, and urine samples is performed to explore systemic metabolic signatures associated with gluten exposure.
The primary objective is to identify gluten-induced transcriptional and cellular changes in peripheral immune cells between baseline (Day 1) and post-intervention (Day 8). Secondary objectives include: (1) assessing dose-dependent immune activation; (2) identifying immune cell subsets and gene expression programs associated with different gluten quantities; (3) characterizing TCR repertoire alterations following gluten exposure; (4) evaluating inter-individual variability in immune responses; (5) analyzing exosomal microRNA changes; and (6) identifying metabolomic signatures associated with gluten ingestion.
研究设计
- 研究类型
- Interventional
- 分配方式
- Randomized
- 干预模型
- Parallel
- 主要目的
- Basic Science
- 盲法
- Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)
盲法说明
Quadruple-blind (participant, care provider, investigator, outcomes assessor). Only three designated team members not involved in routine clinical assessment or outcome evaluation have access to the randomization list in order to prepare the gluten/placebo doses and monitor safety. Data entry and statistical analyses are performed using a blinded database.
入排标准
- 年龄范围
- 8 Years 至 14 Years（Child）
- 性别
- All
- 接受健康志愿者
- 否
入选标准
- •Age 8 to 14 years at study entry.
- •Diagnosis of celiac disease according to ESPGHAN 2020 criteria.
- •At least 18 months on a strict gluten-free diet (GFD).
- •Adequate adherence to the GFD, demonstrated by negative fecal gluten immunogenic peptides (GIP) prior to inclusion.
- •Asymptomatic from a gastrointestinal perspective in the preceding weeks.
- •Ability to swallow the gluten/placebo preparation.
- •Written informed consent from parents/legal guardians and assent from the child.
排除标准
- •Obesity defined as BMI ≥ 95th percentile according to WHO criteria.
- •Diagnosed inflammatory bowel disease or diabetes mellitus.
- •Acute infectious illness at the time of inclusion.
- •Chronic hepatic, pulmonary, renal, or rheumatologic disease.
- •History of severe acute reactions to accidental gluten ingestion.
- •Use of oral corticosteroids or immunosuppressive therapy in the previous 3 months.
- •Any condition that, in the opinion of the investigators, may contraindicate participation or compromise study integrity.
研究组 & 干预措施
Placebo Group
Participants receive a placebo preparation once daily on Days 1, 2, and 3. Blood samples are collected on Day 1 (baseline) and Day 8 (post-intervention). Participants and evaluators remain blinded to allocation.
干预措施: Placebo (Dietary Supplement)
Placebo Group
Participants receive a placebo preparation once daily on Days 1, 2, and 3. Blood samples are collected on Day 1 (baseline) and Day 8 (post-intervention). Participants and evaluators remain blinded to allocation.
干预措施: Collection of blood, stool and urine samples on Day 1 and Day 8. (Procedure)
Low-Dose Gluten (50 mg)
Participants receive 50 mg of gluten once daily on Days 1, 2, and 3, simulating accidental low-level exposure. Blood samples are collected on Day 1 (baseline) and Day 8 (post-intervention). Participants and evaluators remain blinded to allocation.
干预措施: Gluten 50 mg (Dietary Supplement)
Low-Dose Gluten (50 mg)
Participants receive 50 mg of gluten once daily on Days 1, 2, and 3, simulating accidental low-level exposure. Blood samples are collected on Day 1 (baseline) and Day 8 (post-intervention). Participants and evaluators remain blinded to allocation.
干预措施: Collection of blood, stool and urine samples on Day 1 and Day 8. (Procedure)
High-Dose Gluten (5 g)
Participants receive 5 g of gluten once daily on Days 1, 2, and 3, simulating a dietary transgression. Blood samples are collected on Day 1 (baseline) and Day 8 (post-intervention). Participants and evaluators remain blinded to allocation.
干预措施: Gluten 5 g (Dietary Supplement)
High-Dose Gluten (5 g)
Participants receive 5 g of gluten once daily on Days 1, 2, and 3, simulating a dietary transgression. Blood samples are collected on Day 1 (baseline) and Day 8 (post-intervention). Participants and evaluators remain blinded to allocation.
干预措施: Collection of blood, stool and urine samples on Day 1 and Day 8. (Procedure)
结局指标
主要结局
Change in peripheral blood immune cell gene expression after controlled gluten ingestion
时间窗: Day 1 to Day 8
Change from baseline (Day 1) to post-intervention (Day 8) in gene expression profiles of peripheral blood immune cells measured by single-cell RNA sequencing (scRNA-seq).
Change in peripheral blood T-cell receptor repertoire after controlled gluten ingestion
时间窗: Day 1 to Day 8
Change from baseline (Day 1) to post-intervention (Day 8) in T-cell receptor (TCR) diversity and clonality in peripheral blood T cells measured by TCR sequencing
次要结局
- Dose-dependent changes in immune gene expression after gluten exposure(Day 1 to Day 8)
- Dose-dependent changes in T-cell receptor repertoire clonality after gluten exposure(Day 1 to Day 8)
- Inter-individual variability in immune gene expression response to gluten exposure across participants(Day 1 to Day 8)
- Changes in serum exosomal microRNA expression after gluten exposure(Day 1 to Day 8)
- Changes in metabolomic profiles after gluten exposure(Day 1 to Day 8)
- Changes in peripheral blood immune cell type proportions after gluten exposure(Day 1 to Day 8)