Aggregate Metabolic Phenotypes for (Poly)Phenols: Development of an Oral (Poly)Phenol Challenge Test (OPCT)

Study Overview
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Brief Summary
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The study is a single-dose acute clinical trial aiming at identifying aggregate metabolic phenotypes for the main dietary (poly)phenols and assessing the factors associated with their formation. The treatment consists of a nutritional challenge representative of the consumption of (poly)phenols in Europeans (so-called oral (poly)phenol challenge test, OPCT) and foresees the supplementation of three standardized tablets rich in (poly)phenols, prepared from various commercially available plant extracts constituting sources of specific (poly)phenols. Urinary excretion of (poly)phenol metabolites will be evaluated at 24 hours after tablet consumption or, for two subgroups of volunteers, at different time points for 24 hours upon tablet consumption. Blood pressure and heart rate will also be measured and anthropometric data collected. Information will be collected on genetic polymorphisms related to the metabolism of (poly)phenols, gene expression, standard cardiometabolic health biomarkers, cardiometabolic risk scores and gut microbiota profile, through the collection of urine, blood and stool samples. Volunteers will follow a (poly)phenol-free diet before and after the OPCT. To check compliance with food restrictions, a 24-hour recall will be carried out on each visit. For a sub-group of 50 subjects, 3 months after the first challenge, the OPCT will be repeated with further urinary and fecal collection.

Official Title
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Identification of Aggregate Metabolic Phenotypes for the Main Dietary (Poly)Phenols and Assessment of the Factors Associated With Their Formation: Development of an Oral (Poly)Phenol Challenge Test (OPCT)

Conditions
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Individual Variability in (Poly)Phenol Metabolism, Cardiometabolic Health

Intervention / Treatment
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* Dietary Supplement: Oral (poly)phenol challenge test (OPCT)


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Adult (18-74 y) BMI 18.5-35 kg/m^2 Exclusion Criteria: Past cardiovascular events and metabolic diseases including diabetes Inflammatory bowel diseases or gastro-intestinal surgery Renal (GFR<60 ml/min) or hepatic diseases (liver enzymes >2.5 fold higher) Immunodeficiency or autoimmune diseases (other than well-compensated hypothyroidism) Mental disorders Antibiotic therapy within the last month Food allergies Pregnancy or lactation

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 74 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Screening
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: (Poly)phenol tablets<br>Single ingestion of 3 tablets containing different amounts of the most representative dietary (poly)phenols (i.e. flavonoid subclasses, phenolic acids, lignans, ellagitannins, stilbenes, flavonols, procyanidins and phenylethanoids) | Dietary Supplement: Oral (poly)phenol challenge test (OPCT)<br>* Nutritional challenge with standardized (poly)phenol-rich tablets<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Identification of aggregate phenolic metabotypes | Assessing the variability in the urinary excretion of phenolic metabolites among volunteers after consumption of (poly)phenol-rich tablets by using data-driven clustering. | 24 hours post-consumption | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Assessing common cardiometabolic health biomarkers in blood samples | Samples will be processed for the analysis of common biomarkers of cardiometabolic health: total cholesterol (mg/dL), HDL-cholesterol (mg/dL), triglycerides (mg/dL), glucose (mg/dL), insuline (uUI/mL). Analyses will follow standardised routine procedures. | Baseline | 
| Assessing risk prediction scores | Risk prediction scores (i.e., Framingham General Cardiovascular Risk Score, Framingham Heart Study Primary Risk Functions for heart disease, stroke, diabetes, fatty liver disease, and hypertension, Atherosclerotic Cardiovascular Disease (ASCVD) Risk, QRISK3®, QDScore®, Finnish Diabetes Risk Score (FINDRISC)) will be assessed to understand their relationship with the aggregate phenolic metabotypes observed. The higher the scores, the worse the risk of developing the disease. | Baseline | 
| Evaluating trimethylamine N-oxide (TMAO) in urine and plasma samples | TMAO will be quantified in baseline urine and fasting plasma samples by UHPLC-MS/MS. | Baseline | 
| Evaluating eicosanoids in urine samples | Eicosanoids, including prostaglandins, thromboxanes, leukotrienes, isoprostanes and neuroprostanes will be evaluated in baseline urine samples (0-h) by UHPLC-QqQ-MS/MS. | Baseline | 
| Assessing DNA oxidation catabolites and branched fatty acyl esters of hydroxyl fatty acids (FAHFAs) in plasma samples | DNA oxidation catabolites and FAHFAs will be measured in fasting plasma samples by UHPLC-QqQ-MS/MS. | Baseline | 
| Determining genetic differences among subjects | Genotyping will be conducted using genome wide, SNP array approach untargeted methodology, using commercially available SNP arrays and a tSNP approach. This approach will involve the genotyping of approximately 300 SNPs. Genomic DNA will be prepared from PBMCs isolated from blood samples. | Baseline | 
| Assessing transcriptomic signatures in peripheral blood mononuclear cells (PBMCs). | Specific patterns of gene expression related to each metabotype will be investigated in PBMCs by using a microarray-based approach. Analysis will be carried out in a subset of 10 samples for each metabotype. | Baseline | 
| Determining gut microbiota composition and functionality in fecal samples | Microbial profiling will be assessed by shallow shotgun metagenomics. Full shotgun metagenomics analysis will be carried out to determine functional pathways in a sample subset (50 samples). | Baseline | 
| Assessing dietary habits | Dietary habits will be assessed through a food frequency questionnaire. | Baseline | 
| Assessing anthropometric measurements | Weight and height will be combined to report BMI in kg/m^2 and this will be carried out according to standardized procedures. Waist circumference and hip circumference, waist-to-height ratio, waist-to-hip ratio, and body composition measurement (skinfold test and bioelectrical impedance analysis). | Baseline | 
| Assessing blood pressure and heart rate | Systolic and diastolic blood pressure and heart rate of each volunteer will be obtained after a 5-min rest in a seated position in the baseline visit. | Baseline | 
| Evolution over the time of the phenolic metabolites in urine samples | Assessed by using UHPLC-MS/MS for individual detection and quantification. | Different collection times for 24 hours (0; 0-3; 3-6; 6-9; 9-12; 12-24; 24 h) | 
| Untargeted urinary metabolomics | The untargeted LC-IMS-MS metabolomics approach will allow to assess potential differences among the metabolomes of individuals belonging to different aggregate phenolic metabotypes. | Baseline and 24 hours post-consumption | 
| Assessing the stability of aggregate phenolic metabotypes among individuals after 3 months | Assessing the variability in the urinary excretion of phenolic metabolites among volunteers after consumption of (poly)phenol-rich tablets, considering the metabotype to which each volunteer belongs to once the volunteer re-do the oral (poly)phenol challenge test after 3 months from the first supplementation. | 24 hours post-consumption in a test carried out after 3 months after the first supplementation | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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metabolic phenotypes, intervention study, (poly)phenols, metabotypes, cardiometabolic diseases, inter-individual variability, phenolic metabolites