Dietary Proteins: Metagenomic and Metabolomics Approaches for Human Biomarkers Identification

NCT05611138 | Completed

• 1 other identifier: PanProtein 2
• Study Type: interventional
• Target: 59
• Locations: 1 country
• Sites: 1

Brief Summary

This project aims to create a methodological framework, including the discovery and validation of novel biomarker panels to decipher the impact of plant-based protein intake on health biomarkers. Moreover, the possibility of predicting biomarkers production will be tested by a colonic in vitro fermentation study using study participants' faecal samples.

Conditions

Healthy

Outcome Measures

Primary Outcomes (7)

• Changes in the faecal metabolites using targeted and untargeted metabolomics during the intervention

  Difference in faecal metabolites depending on the protein supplementation amount and from colonic in vitro fermentation. Targeted and untargeted metabolomics measurements will be used (e.g., short-chain fatty acids and branched chain fatty acids analyses, and polar and non polar metabolomics). Faecal samples will have their metabolites extracted and quantified by liquid chromatography coupled with high-resolution time-of-flight mass spectrometry (UHPLC-qToF-MS), and gas chromatography coupled with high-resolution mass spectroscopy (GC-Orbitrap). The level of metabolites will be compared throughout the whole study.

  8 weeks

• Changes in the plasma metabolites using targete and untargeted metabolomics during the intervention

  Difference in plasma metabolites (blood obtained fastened state) depending on the protein supplementation amount. Targeted and untargeted metabolomics measurements will be used (e.g., lipids, and polar and non polar metabolomics). Plasma samples will have their metabolites extracted and quantified by liquid chromatography coupled with high-resolution time-of-flight mass spectrometry (UHPLC-qToF-MS), and gas chromatography coupled with high-resolution mass spectroscopy (GC-Orbitrap). The level of metabolites will be compared throughout the whole study.

  4 weeks

• Profile/composition of the gut microbiota during the intervention

  Difference in the gut microbiota profile/composition because of the protein supplementation and from colonic in vitro fermentation will be assessed by 16s/NGS. Alpha and Beta diversity will be measured and compared throughout the 4 weeks of intervention and baselines. In the case of NGS, the genome sequence will be annotated and the resulting set of genes encoding for metabolic enzymes will be extracted. Data from shotgun metagenomics and metabolomics will be integrated by constructing dependency networks, with a special attempt to infer causal relationships among different variables.

  8 weeks

• Routinely analysed markers for protein intake/compliance during the intervention - urine

  Difference in 24 h urine samples markers due to the protein supplementation (e.g., for 24 h urine: urea nitrogen from 24 h urine, creatinine, uric acid)

  4 weeks

• Routinely analysed markers for protein intake/compliance during the intervention - blood

  Difference in blood (serum) samples markers due to the protein supplementation (e.g., urea and creatinine)

  4 weeks

• Changes in the plasma metabolites using targeted and untargeted metabolomics during the dietary challenge

  Difference in plasma metabolites depending on the protein taken as breaskfast. Targeted and untargeted metabolomics measurements will be used (e.g., polar and non polar metabolomics). Plasma samples will have their metabolites extracted and quantified by liquid chromatography coupled with high-resolution time-of-flight mass spectrometry (UHPLC-qToF-MS), and gas chromatography coupled with high-resolution mass spectroscopy (GC-Orbitrap). The level of metabolites will be compared with those found in the fastening state.

  4 weeks

• Colonic in vitro fermentation

  Subjects will have their fresh faecal samples used in the colonic in vitro fermentation experiment to evaluate whether and how gut metabolites production can be predicted in vitro. The same proteins used in the dietary intervention study will be digested in vitro and used for the batch in vitro colonic fermentation. Supernatant will be used for metabolomics analysis (outcome 1) and selected fermentation time point pellets will be used for microbiota composition and functional analyses (outcome 3).

  4 weeks

Secondary Outcomes (18)

• Food intake using food diaries

  8 weeks

• Assessment of the Gastrointestinal Symptom rating scale (GSRS) during the study

  8 weeks

• Assessment of the bowel movement using the Bristol scale diary

  8 weeks

• Assessment of physical activity level

  8 weeks

• Assessment of height

  4 weeks

Study Arms (3)

Isolated pea protein

EXPERIMENTAL

Dietary supplementation with isolated pea protein powder. Amount: 0.5 g of protein from the supplement per kilogram of body weight, taken in 3 portions during the day, for 4 weeks.

Dietary Supplement: Protein supplementation

Isolated whey protein

EXPERIMENTAL

Dietary supplementation with isolated whey protein. Amount: 0.5 g of protein from the supplement per kilogram of body weight, taken in 3 portions during the day, for 4 weeks.

Dietary Supplement: Protein supplementation

Concentrated pea protein

EXPERIMENTAL

Dietary supplementation with concentrated pea protein Amount: 0.5 g of protein from the supplement per kilogram of body weight, taken in 3 portions during the day, for 4 weeks.

Dietary Supplement: Protein supplementation

Interventions

Protein supplementation DIETARY_SUPPLEMENT

Participants will be instructed to dissolve the protein powder in liquids (such as water) or on their food (e.g., yoghurt, oat porridge). A subset of the participants (max. 15 per arm) for visits 2 and 6 will be invited to participate in a postprandial challenge, where they will receive the daily protein portion as breakfast.

Concentrated pea protein; Isolated pea protein; Isolated whey protein

Eligibility Criteria

• Age: 18 Years - 45 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)

You may qualify if:

• Age 18-45 years
• Body mass index (BMI) 18,5-30 kg/m2
• Weight stable within the previous 3 months
• Maintenance of the usual physical activity habits during the study
• Intake of fibre between 15 and 25 g per day (evaluated by 3 food diaries or 24-hours recalls)
• Omnivores

You may not qualify if:

• Acute chronic disease, inflammatory or functional gastrointestinal diseases and any other disease or disorder that could affect the outcome of the study
• Use of a medication that may interfere the study outcome
• Eating disorder
• High protein intake (more than 15% of energy or maximum 1,2 g of protein per kg of body weight per day; evaluated by 3 food diaries or 24-hours recalls)
• Use of antibiotic medication during the last 3 months prior the first visit
• Use of antibiotic medication very early in life (e.g., asthmatic children or ear inflammation)
• Use of laxative or anti-diarrhoea medication within the past 3 months before the study
• Regular consumption of probiotic or prebiotic product for the past 6 weeks before the study
• Special diet that is considered to affect the study participation and/or study results, for example, high protein diets
• More than 5 h of moderate-vigorous exercise per week
• Pregnancy or breastfeeding
• Intolerance to dietary supplements that will be used in the study
• Smoking
• Abuse of alcohol or drugs

Sponsors & Collaborators

• Örebro University, Sweden: lead

Study Sites (1)

Campus USÖ, Örebro University

Örebro, 70362, Sweden

Location

Study Officials

• Robert Brummer, MD, PhD

  Örebro University, Sweden

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: TRIPLE
• Who Masked: PARTICIPANT, INVESTIGATOR, OUTCOMES ASSESSOR
• Purpose: BASIC SCIENCE
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Model Details: Healthy male and female adult subjects (n=87) will be recruited to take part in a double-blinded, randomized, 3-arms parallel trial. Subjects that are eligible to take part in the study will be blinded and randomly allocated to consume an isolated plant protein (arm 1) or isolated animal protein (arm 2) or plant-based source of protein (arm 3). The subjects eligible to take part in this study will consume the proteins for 4 weeks, before the intervention they will have a baseline of two weeks, and after the intervention, they will have more two visits. Faecal and blood samples will be used to answer the main outcome of gut-related metabolites and microbiota profile. Blood and urine samples will be used to measure health status and compliance (protein intake markers). Questionnaires will be used to access gastrointestinal symptoms and bowel movement, physical activity level and dietary intake.

Study Record Dates

• First Submitted: November 2, 2022
• First Posted: November 9, 2022
• Study Start: September 15, 2022
• Primary Completion: April 14, 2023
• Study Completion: April 14, 2023
• Last Updated: April 28, 2023

Record last verified: 2023-04

Locations

SE (1)