Ketone Supplementation, Glucose Control, and Cardiovascular Function

Study Overview
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Brief Summary
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Post-prandial hyperglycemic excursions induce a cascade of deleterious effects on the body, including increased inflammation, production of reactive oxygen species, and impaired cardiovascular function. Ingestion of an exogenous oral ketone supplement blunts hyperglycemia in response to an oral glucose tolerance test. Accordingly, it is hypothesized that exogenous ketone supplement ingestion prior to a meal could be an effective strategy for blunting postprandial hyperglycemia. Therefore, the purpose of this study is to investigate the effect of short-term (14-days) pre-meal exogenous ketone supplementation on glucose control, cardiovascular function, inflammation, and oxidative stress in individuals at an elevated risk of type 2 diabetes.

Official Title
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The Effects of Exogenous Ketone Supplementation on Cardiovascular Function and Glucose Control

Conditions
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Hyperglycemia, Postprandial, Overweight and Obesity, Diabetes Mellitus Risk

Intervention / Treatment
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* Dietary Supplement: Exogenous ketone monoester


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Elevated waist circumference (>102 cm for males, >88 cm for females) and/or Obesity (BMI > 30 kg/m2) and/or Diagnoses of prediabetes based on A1C (5.7-6.4%) and/or fasting plasma glucose (5.6-6.9 mmol/l) using ADA criteria Exclusion Criteria: Competitively trained endurance athlete Actively attempting to lose weight History of mental illness or existing neurological disease(s) Previous cardiovascular events (i.e., heart attack, stroke) Diagnoses of diabetes Hypoglycemia Irritable bowel syndrome or inflammatory bowel disease Taking medication that may interfere with insulin sensitivity Currently following a ketogenic diet or taking ketone supplements Unable to commit for 2 separate 14-day trials and unable to follow a controlled diet

Ages Eligible for Study
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Minimum Age: 30 Years
Maximum Age: 69 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Prevention
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Experimental<br>Participants will consume 20 g of an active oral exogenous ketone monoester supplement 15 minutes prior to each meal of the day for a 14-day period. Pre-intervention (baseline) and post-intervention measurements will be obtained before and immediately after the 14-day period. All meals will be provided throughout the supplementation period Participants will wear a continuous glucose monitor for 6 consecutive days during the supplementation period. | Dietary Supplement: Exogenous ketone monoester<br>* Participants will consume 20g of the oral ketone monoester supplement 15 minutes prior to each meal of the day for 14 days. All meals will be provided throughout the 14-day supplementation period.<br>* Other names: HVMN ketone monoester supplement;|
| Placebo Comparator: Placebo<br>Participants will consume a flavor matched placebo drink and undergo the same procedures described in the Experimental Arm | Dietary Supplement: Exogenous ketone monoester<br>* Participants will consume 20g of the oral ketone monoester supplement 15 minutes prior to each meal of the day for 14 days. All meals will be provided throughout the 14-day supplementation period.<br>* Other names: HVMN ketone monoester supplement;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Glucose control | Post-prandial glucose excursions will be measured by continuous glucose monitoring using the iPro2 CGM by Medtronic in both the active and placebo supplement conditions. Post-prandial glucose following breakfast, lunch, and dinner will be averaged together. | 2 hours after a meal | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change from baseline flow mediated dilation at 14 days | Vascular function will be assessed by flow mediated dilation of the brachial artery using vascular ultrasound. A cuff will affixed on the forearm, distal to the brachial artery and will be inflated for 5 minutes. Flow mediation dilation will be measured over a 3-minute period following cuff release. | Day 0 (Pre-intervention) and Day 14 (post-intervention) | 
| Change from baseline histone acetylation at 14 days | Histone H3 acetylation status will be quantified by flow cytometry using conjugated acetyl-histone H3 antibody specific for Lys9 (Pacific Blue 445) and the conjugated acetyl-histone H3 antibody specific for Lys14 (Alexa Fluor 488). | Day 0 (Pre-intervention) and Day 14 (post-intervention) | 
| Change from baseline mitochondrial superoxide production at 14 days | Oxidative Stress will be measured by mitochondrial superoxide production in blood lymphocytes, monocytes, and neutrophils by flow cytometry using the MitoSOX red assay (ThermoFisher #M36008) and total intracellular ROS via the DCFDA assay (Sigma #D6883) | Day 0 (Pre-intervention) and Day 14 (post-intervention) | 
| Change from baseline cognition (executive functions) at 14 days | Cognition will be assessed using a customized battery of psychometrically validated tests within the domain of executive functions using the iPad-based app BrainBaseline. The tests will be the Stroop test, task-switching test, digit-symbol substitution test, and the n-back test. | Day 0 (Pre-intervention) and Day 14 (post-intervention) | 
| Change from baseline plasma glucose at 14 days | Venous blood samples will be taken and plasma glucose will be measured using a hexokinase method. | Day 0 (Pre-intervention) and Day 14 (post-intervention) | 
| Change from baseline plasma insulin at 14 days | Venous blood samples will be taken and plasma insulin will be measured using a high-sensitivity human insulin ELISA. | Day 0 (Pre-intervention) and Day 14 (post-intervention) | 
| Change from baseline plasma free fatty acids at 14 days | Venous blood samples will be taken and free fatty acids will be measured by colorimetric assay. | Day 0 (Pre-intervention) and Day 14 (post-intervention) | 
| Change from baseline interleukin-1(IL)-1beta at 14 days | Mature IL-1beta secretion will be quantified by ELISA run in duplicate | Day 0 (Pre-intervention) and Day 14 (post-intervention) | 
| Change from baseline caspase-1 activation at 14 days | Caspase-1 activation will be quantified by flow cytometry. The fluorescent inhibitor probe FAM-YVAD-FMK binds covalently to activated caspase-1 and emits at 530nm | Day 0 (Pre-intervention) and Day 14 (post-intervention) | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Flow mediated dilation, Glucose control, Inflammation, Immune cell function, Cognition, Oxidative stress, Cardiovascular function