Choline Source, Gut Microbiota and Trimethylamine-N-oxide Response

Effect of Choline Source and Gut Microbiota Composition on Trimethylamine-N-oxide Response in Humans

The purpose of this research is to determine the production of trimethylamine-N-oxide (TMAO) from different forms of choline and whether this response is modified by the gut microbiota composition.

Study Overview

• Status: Completed
• Conditions: Metabolism
• Intervention / Treatment: Other: Water-soluble choline; Other: Fat-soluble choline; Other: No choline control

Detailed Description

The overall goal of this research is to identify dietary and physiological factors contributing to elevated levels of trimethylamine-N-oxide (TMAO), a choline-derived gut-microbiome-dependent metabolite that has been identified to increase cardiovascular disease risk. Our recent findings indicate that the gut microbiome may account for variations in TMAO levels, whereby those with a greater enrichment of Firmicutes to Bacteroidetes had elevated TMAO response to dietary precursor intake. However, the interaction between choline intake and gut microbiota composition as a determinant of interindividual variations in TMAO response has not been investigated. This study sought to i) compare plasma and urinary TMAO response after acute challenge containing different forms of choline; and ii) to determine the association between differences in TMAO response with differences in gut microbiota composition. To accomplish these objectives, a randomized, controlled cross-over study was conducted in healthy participants (n=41). The study incorporated three arms comprised of study meals containing (i) 600 mg choline as choline bitartrate; (ii) 600 mg choline as phosphatidylcholine; or (iii) no choline. Each meal was served with a bagel with margarine-butter spread and one cup of water, administered in a single day and separated by a 1-week washout period. Baseline blood sample was obtained by a phlebotomist using a standard venipuncture procedure, and participants collected their baseline urine sample. They also turned in a one-time self-collected baseline stool sample. Following the consumption of the study meal, serial blood samples were collected at 30 min and 1, 2, 4 and 6 h, and urine samples collected throughout the 6 h study period. At 4.5 h, participants were provided a fixed fruit snack (i.e., 2 single serving prepackaged applesauce) and water.

• Study Type: Interventional
• Enrollment (Actual): 44
• Phase: Not Applicable

Contacts and Locations

Study Locations

• United States
  • Utah
    • Logan, Utah, United States, 84322
      • Center for Human Nutrition Studies Clinic, Utah State University

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 17 years to 46 years (Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

Description

Inclusion Criteria:

• Healthy men and women of any race or ethnicity
• Age 21-50 y
• BMI 20-24.9 kg/m2 or BMI 30-39.9 kg/m2

Exclusion Criteria:

• Age > 50 y
• BMI outside of the normal-weight or obese range (BMI < 20 kg/m2; BMI 25-29.9 kg/m2; or BMI > or = 40 kg/m2)
• Pregnant or planning to become pregnant during the course of the study; currently breastfeeding (females)
• Vegetarians
• Smokers or recreational drug users
• Individuals with gastrointestinal diseases or complaints, chronic illnesses or other metabolic diseases (including trimethylaminuria)
• Individuals who have taken antibiotics within the past 2 months
• Individuals who are not willing to discontinue pre- and probiotics and dietary supplements for the time leading up to 2 months before the study and during the study

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Randomized
• Interventional Model: Crossover Assignment
• Masking: Double

Number of Arms

3

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Water-soluble choline; A breakfast meal consisting of 1 cup of tomato soup containing 600 mg choline as choline bitartrate; served with a bagel with margarine-butter spread and one cup of water.	Other: Water-soluble choline; 600 mg choline as choline bitartrate
Experimental: Fat-soluble choline; A breakfast meal consisting of 1 cup of tomato soup containing 600 mg choline as phosphatidylcholine; served with a bagel with margarine-butter spread and one cup of water.	Other: Fat-soluble choline; 600 mg choline as phosphatidylcholine
Active Comparator: No choline; A breakfast meal consisting of 1 cup of tomato soup containing no choline; served with a bagel with margarine-butter spread and one cup of water.	Other: No choline control; No choline

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
TMAO metabolite concentration change	Plasma TMAO metabolite response	Blood: study baseline, 30 minutes and 1 hour, 2 hours, 4 hours and 6 hours
TMAO metabolite concentration change	Urinary TMAO metabolite response	Urine: study baseline, pooled 6 hours study period
Gut microbiome profile	16S rRNA	Stool: one-time baseline

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
One-carbon metabolite concentration change	Plasma choline metabolite response	Blood: study baseline, 30 minutes and 1 hour, 2 hours, 4 hours and 6 hours
Phosphatidylcholine concentration change	Plasma phosphatidylcholine response	Blood: study baseline, 30 minutes and 1 hour, 2 hours, 4 hours and 6 hours
One-carbon metabolite concentration change	Urinary choline metabolite response	Urine: study baseline, pooled 6 hours study period
Inflammation and cardiovascular disease risk marker concentration change	Plasma TNF-α and IL-6	Blood: study baseline to 6 hours
Flavin monooxygenase 3 (FMO3) 472 G>A genotype variant	Genetic polymorphism	Blood: study baseline

Collaborators and Investigators

• Sponsor: Utah State University
• Investigators: Principal Investigator: Clara E Cho, PhD, Utah State University

Publications and helpful links

General Publications

• Wang Z, Klipfell E, Bennett BJ, Koeth R, Levison BS, Dugar B, Feldstein AE, Britt EB, Fu X, Chung YM, Wu Y, Schauer P, Smith JD, Allayee H, Tang WH, DiDonato JA, Lusis AJ, Hazen SL. Gut flora metabolism of phosphatidylcholine promotes cardiovascular disease. Nature. 2011 Apr 7;472(7341):57-63. doi: 10.1038/nature09922.
• Tang WH, Wang Z, Levison BS, Koeth RA, Britt EB, Fu X, Wu Y, Hazen SL. Intestinal microbial metabolism of phosphatidylcholine and cardiovascular risk. N Engl J Med. 2013 Apr 25;368(17):1575-84. doi: 10.1056/NEJMoa1109400.
• Cho CE, Taesuwan S, Malysheva OV, Bender E, Tulchinsky NF, Yan J, Sutter JL, Caudill MA. Trimethylamine-N-oxide (TMAO) response to animal source foods varies among healthy young men and is influenced by their gut microbiota composition: A randomized controlled trial. Mol Nutr Food Res. 2017 Jan;61(1). doi: 10.1002/mnfr.201600324. Epub 2016 Aug 3.

Study record dates

Study Major Dates

• Study Start (Actual): November 9, 2017
• Primary Completion (Actual): June 16, 2018
• Study Completion (Actual): July 25, 2020

Study Registration Dates

• First Submitted: January 22, 2020
• First Submitted That Met QC Criteria: February 3, 2020
• First Posted (Actual): February 5, 2020

Study Record Updates

• Last Update Posted (Actual): November 4, 2020
• Last Update Submitted That Met QC Criteria: November 3, 2020
• Last Verified: November 1, 2020

More Information

Terms related to this study

• Keywords: Gut microbiota; Metabolism; Dietary precursor
• Additional Relevant MeSH Terms: Molecular Mechanisms of Pharmacological Action; Antimetabolites; Gastrointestinal Agents; Hypolipidemic Agents; Lipid Regulating Agents; Nootropic Agents; Lipotropic Agents; Choline
• Other Study ID Numbers: 8547

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: No

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No