Mitochondrial Effects of C18:0 Supplementation in Humans

Study Overview
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Brief Summary
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The purpose of this crossover study is to determine whether nutritional supplementation of C18:0 in humans has mitochondrial effects as shown in Drosophila and human cell culture. We will compare a study cohort of patients with diagnosed type 2 diabetes with non-diabetics. Participants will undergo a 2-day low-fat vegan diet and will then be supplemented with a bolus of C18:0. Changes in the mitochondrial morphology and function of white blood cells will be scored by immunofluorescence and FACS analysis.

Detailed Description
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The purpose of this study is to determine whether nutritional supplementation of C18:0 in humans has mitochondrial effects as shown in Drosophila and human cell culture. We will compare a study cohort of patients with diagnosed type 2 diabetes with non-diabetics. Participants will undergo a 2-day low-fat vegan diet to reach baseline levels of C18:0 and will then be fed a milkshake supplemented with 24g of C18:0, which corresponds roughly to the C18:0 content of a fast-food meal. Blood samples will be taken at baseline and several hours after intake. We will look at changes in mitochondrial morphology of neutrophils by immunofluorescence, and score mitochondrial function via FACS analysis. Since this study is designed as a crossover study, participants will also receive a mock milk shake after another 2 days of low-fat vegan diet.

Official Title
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Mitochondrial Effects of C18:0 Supplementation in Type 2 Diabetics Versus Healthy Controls

Conditions
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Alteration of Mitochondrial Membrane, Type2 Diabetes, Fatty Acid Deficiency

Intervention / Treatment
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* Dietary Supplement: C18:0
* Dietary Supplement: mock


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: type 2 diabetes, either dietary treatment or oral medication must be able to give consent Exclusion Criteria: insulin treated diabetes mellitus severe diseases inducing wasting (e.g. cancer, liver cirrhosis, renal failure) conditions of malnourishment severe anemia pregnancy alcohol abuse

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: Crossover Assignment
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Non-diabetics<br>Non-diabetic volunteers with HbA1c < 6.5%. Subjects will be treated with C18:0 supplementation or mock. | Dietary Supplement: C18:0<br>* Receives 24g of C18:0 in a low-fat banana milkshake.<br>* Other names: stearic acid, Sigma-Aldrich, product number W303518;Dietary Supplement: mock<br>* Low fat banana milkshake without C18:0 supplement.<br>|
| Experimental: Type 2 Diabetics<br>Type 2 Diabetes according to common definitions, but we exclude insulin-treated Type 2 diabetics because nutritional intervention is more difficult/risky. Subjects will be treated with C18:0 supplementation or mock. | Dietary Supplement: C18:0<br>* Receives 24g of C18:0 in a low-fat banana milkshake.<br>* Other names: stearic acid, Sigma-Aldrich, product number W303518;Dietary Supplement: mock<br>* Low fat banana milkshake without C18:0 supplement.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in Mitochondrial Morphology | Mitochondria of neutrophils are stained and scored via immunofluorescence microcsopy, either as fragmented, intermediate or fused. Statistical calculations will be performed on changes in fragmentation status after treatment. | 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h | 
| Changes in Mitochondrial Function | Mitochondrial membrane potential and ROS production in neutrophils will be analyzed via FACS. Statistical calculations will be performed on changes in the respective levels after treatment. | on the day of supplementation at 0, 3 and 6 h | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| plasma iron, transferrin, ferritin, ferroportin and hepcidin levels | Measurement of iron, transferrin, ferritin, hepcidin and ferroportin from serum at all timepoints via ELISA. Changes in plasma levels will be correlated to primary endpoints. | 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h | 
| plasma methylglyoxal levels | Methylglyoxal levels in plasma analyzed via liquid chromatography-mass spectrometry. Changes will be correlated to primary endpoints. | 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h | 
| plasma fatty acid levels | Fatty acids along with other lipid parameters like triglycerides and cholesterol for normalization purposes will be measured. | 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h | 
| insulin resistance | Insulin and glucose levels will be measured at each time point, HOMA index will be calculated. | 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h | 
| diabetic late complications | Patients with confirmed HbA1c > 6,5% will be considered diabetic. Then albumin in urine will be measured and diabetic neuropathy will be assessed clinically via NDS/NSS scoring. | 2 days before supplementation | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Transferrin Receptor, Type 2 Diabetes, Stearic Acid (C18:0), Mitochondrial Morphology, Crossover Study