Effects of Choline From Eggs vs. Supplements on the Generation of TMAO in Humans

Study Overview
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Brief Summary
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The investigators are interested in learning more about choline, a nutrient required by the body. The body does make some choline, but it does not make enough to support health and the rest must be acquired through diet. Eggs, and especially egg yolks, are a major dietary source of choline. Choline can also be given as a dietary supplement. Ingestion of choline supplements has been linked to an increased concentration of a compound called TMAO (trimethylamine N-oxide). Elevated TMAO levels have been linked to higher heart disease risk. With this study, the investigators hope to learn whether there is a difference in the way your body responds to the ingestion of a choline supplement versus the choline found within eggs.

Detailed Description
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The principal goal for the study is to examine whether there is a difference between the ingestion of choline through supplements versus choline found within eggs on plasma TMAO levels. The investigators have previously shown that dietary intake of trimethylamines, including the choline group of phosphatidylcholine (PC), is mechanistically linked to cardiovascular disease risk and that the metabolism of these trimethylamine nutrients in humans is modulated by the intestinal microbes (gut microbes). Additionally, extensive animal studies link an essential role of gut microbiota to the metabolism of choline and the production of metabolites that promote / accelerate atherosclerotic processes. The investigators have also recently shown a 10-fold increase in plasma TMAO levels following supplementation with choline bitartrate supplements. However, another pilot study by a collaborator (unpublished) did not show the same increase in plasma TMAO levels following the ingestion of whole eggs, a major dietary source of choline. Therefore, with this study the investigators wish to examine the differences, if any, between the ingestion of an equivalent mass of total choline in the free form (as bitartrate salt) as a supplement vs. within whole eggs. Eggs, and specifically the egg yolk, contain a large amount of total choline. However, egg white contains potential anti-microbial peptides that could influence gut microbial composition and function, and therefore impact conversion of choline into TMA and TMAO observed in subjects. Therefore, the investigators hypothesize that the consumption of whole eggs (hardboiled) will not elevate plasma TMAO levels to the same extent as a comparable amount of total choline ingested in capsule form as the choline bitartrate salt. The investigators further hypothesize that the consumption of egg white with choline bitartrate tablets may result in less of a rise in TMAO levels than ingestion of the choline bitartrate supplement alone.

Official Title
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Effects of Choline From Eggs vs. Supplements on the Generation of TMAO in Humans (EGGS)

Conditions
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Cardiovascular Risk Factor

Intervention / Treatment
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* Dietary Supplement: Choline Bitartrate
* Other: Pre-cooked, pre-peeled whole hardboiled eggs
* Other: Egg whites from pre-cooked, pre-peeled hardboiled eggs
* Dietary Supplement: Phosphatidylcholine capsules


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Men and women age 18 years or above. Willing to remain on aspirin or stay off aspirin or aspirin products for 1 week prior to starting the study and throughout the study period. Able to provide informed consent and comply with study protocol. Able to be off all other supplements during the study period. Exclusion Criteria: Significant chronic illness. Active infection or received antibiotics within 1 month of study enrollment. Use of over-the-counter probiotic within the past month Chronic gastrointestinal disorders, such as ulcerative colitis or Crohn's disease. Allergy to eggs or lactose. Having undergone bariatric procedures or surgeries such as gastric banding or bypass. Pregnancy. Any condition that, in the judgment of the Investigator, would place a patient at undue risk by being enrolled in the trial or cause inability to comply with the trial.

Ages Eligible for Study
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Minimum Age: 18 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: Basic Science
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Whole Hardboiled Eggs<br>Subjects will consume four (4) pre-cooked, pre-peeled whole hardboiled eggs per day for 28 days. | Other: Pre-cooked, pre-peeled whole hardboiled eggs<br>* Obtained from a commercial source.<br>|
| Experimental: Choline Bitartrate Tablets<br>Subjects will consume two (2) 500mg choline bitartrate tablets per day for 28 days. | Dietary Supplement: Choline Bitartrate<br>* 500mg choline bitartrate tablets<br>|
| Experimental: Hardboiled Eggs + Choline Bitartrate Tablets<br>Subjects will consume both four (4) whole, pre-cooked, pre-peeled hardboiled eggs and two (2) 500mg choline bitartrate tablets per day for 28 days. | Dietary Supplement: Choline Bitartrate<br>* 500mg choline bitartrate tablets<br>Other: Pre-cooked, pre-peeled whole hardboiled eggs<br>* Obtained from a commercial source.<br>|
| Experimental: Egg Whites + Choline Bitartrate Tablets<br>Subjects will consume both the egg whites (no yolks) of four (4) pre-cooked, pre-peeled hardboiled eggs and two (2) 500mg choline bitartrate tablets per day for 28 days. | Dietary Supplement: Choline Bitartrate<br>* 500mg choline bitartrate tablets<br>Other: Egg whites from pre-cooked, pre-peeled hardboiled eggs<br>* Egg whites from pre-cooked, pre-peeled hardboiled eggs. The yolks are removed and discarded.<br>|
| Experimental: Phosphatidylcholine Capsules<br>Subjects will consume six (6) 420 mg phosphatidylcholine capsules by mouth per day for 28 days. | Dietary Supplement: Phosphatidylcholine capsules<br>* 420 mg phosphatidylcholine capsules obtained from a commercial source.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in Plasma Levels of Fasting Trimethylamine-N-oxide (TMAO), a Choline Metabolite | Changes in levels of non-labeled TMAO from baseline to end-of-study (day 28) as measured by established techniques by mass spectrometry. | Baseline, 28 days | 
| Changes in Platelet Function With Increased Choline Intake | The activation and functioning of platelets within a single subject will be compared before and after increased choline intake. | Baseline, Day 28 | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in Levels of Fasting Trimethylamine-N-oxide (TMAO) in 24-hour Urine Collections | Changes in levels of non-labeled TMAO from baseline to Day 28 measured by established mass spectrometry techniques. | Baseline, Day 28 | 
| Changes in Plasma Levels of Fasting Choline | Fasting plasma levels of choline from samples obtained at baseline and at day 28 were compared. | Baseline, Day 28 | 
| Changes in Plasma Levels of Fasting Carnitine. | Fasting plasma levels of carnitine from samples obtained at baseline and at day 28 were compared. | Baseline, Day 28 | 
| Changes in Plasma Levels of Fasting Betaine. | Fasting plasma levels of betaine from samples obtained at baseline and at day 28 were compared. | Baseline, Day 28 | 
| Changes in Lipid Profile, Total Cholesterol | Changes in total cholesterol levels between baseline and Day 28 | Baseline, Day 28 | 
| Changes in Lipid Profile, HDL | Changes in measured HDL levels between baseline and Day 28 | Baseline, Day 28 | 
| Changes in Lipid Profile, LDL | Changes in measured LDL levels between baseline and Day 28 | Baseline, Day 28 | 
| Changes in Lipid Profile, Triglycerides | Changes in measured triglyceride levels between baseline and Day 28 | Baseline, Day 28 | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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TMAO, choline, eggs, gastrointestinal microbiome, healthy volunteers