The Effects of Dietary Supplementation on Serum Protein Glycosylation

Study Overview
=================
Brief Summary
-----------------
Two randomized double-blind placebo-controlled clinical studies to assess the effects of dietary glycan monomer supplementation on the immune system, especially antibody glycosylation in healthy adults. Immune profiling and glycoproteomics will be performed on serum isolated at these same time points.

Official Title
-----------------
The Effects of Dietary Supplementation on Serum Protein Glycosylation

Conditions
-----------------
Healthy

Intervention / Treatment
-----------------
* Dietary Supplement: Monosaccharide Powder
* Dietary Supplement: Placebo Comparator


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: Adult men/women (age 18-45) who are in general good health with Body Mass Index (BMI) range of 18.5-25 Exclusion Criteria: Adults younger than 18 or older than 45 years of age Women who are pregnant, actively nursing or have had a pregnancy within the last year Women who are peri-menopausal or post-menopausal Women with irregular menstrual cycles (more frequently than every 21 days or lasts longer than 8 days. Missed, early, or late periods are also considered signs of an irregular cycle) Women on hormonal contraception, including birth control, hormonal intrauterine device, or contraceptive implant Individuals with past medical history of an autoimmune condition or malignancy, excluding non-melanoma skin cancer Individuals with cardiovascular, pulmonary, reproductive, endocrine, metabolic, neurologic, gastrointestinal, hematologic, or infectious diseases of any type Individuals with diagnosis of galactosemia or congenital disorders of glycosylation Individuals with phenylketonuria Individuals currently taking prescription medications, or who have taken prescription medications within the last 3 months Individuals currently taking over-the-counter medications Individuals currently taking or who have taken supplements including herbal, protein or vitamin supplements in the last 3 months (e.g. whey protein, St. John's Wort, green tea supplements, biotin, creatine supplements), excluding multivitamins or essential vitamins Individuals with BMI less than 18.5 or greater than 25 Individuals with prior history of severe food or drug allergic reactions Individuals with allergic reaction or adverse reaction to shellfish, N-acetylglucosamine, galactose, Spirulina/chlorella/algae supplements, or arabinose containing compounds/foods/supplements Individuals with first-degree relative with history of an autoimmune condition Individuals with social history of current use of tobacco, alcohol or other drugs Individuals with dietary restrictions (vegetarians are permitted to participate in the study) or atypical exercise patterns, or to whom any of the following dietary habits or characteristics apply (the following exclusion criteria are placed to minimize variability in diet/exercise patterns in our pilot study population): Actively dieting or trying to lose weight Vegan diet Consume equal to or greater than 2 cups of tea a day Consume equal to or greater than 4 cups of coffee a day Consume equal to or greater than 3 cups of fruit juice a day Practice intensive exercise patterns (marathon training, workouts >4 hours a day) Consume soda or energy drinks of any amount Consume fast food equal to or greater than 5 days per week Consume greater than one serving of alcohol per day (1 serving of wine = 6 oz, 1 serving of beer = 12 oz, 1 serving of spirits = 1 oz) On a carbohydrate-restricted or Paleo diet, or calorie-restricted diet (less than 20-25% of maintenance calories) Adults unable to consent Prisoners

Ages Eligible for Study
-----------------
Minimum Age: 18 Years
Maximum Age: 45 Years

Sexes Eligible for Study
-----------------
All

Accepts Healthy Volunteers
-----------------
Accepts Healthy Volunteers

Study Plan
=================
How is the study designed?
-----------------

Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Experimental Supplement 1<br>6 subjects will consume 12 grams of dietary supplement 1 daily for 4 weeks | Dietary Supplement: Monosaccharide Powder<br>* Monosaccharide powder dissolved in water.<br>|
| Experimental: Experimental Supplement 2<br>6 subjects will consume 12 grams of dietary supplement 2 daily for 4 weeks | Dietary Supplement: Monosaccharide Powder<br>* Monosaccharide powder dissolved in water.<br>|
| Experimental: Experimental Supplement 3<br>20 subjects will consume 50 grams of dietary supplement 3 daily for 4 weeks | Dietary Supplement: Monosaccharide Powder<br>* Monosaccharide powder dissolved in water.<br>|
| Placebo Comparator: Placebo Comparator 1<br>6 subjects will consume 12 grams of placebo dietary supplement daily for 4 weeks | Dietary Supplement: Placebo Comparator<br>* Placebo (glucose powder)<br>|
| Placebo Comparator: Placebo Comparator 2<br>6 subjects will consume 50 grams of placebo dietary supplement daily for 4 weeks | Dietary Supplement: Placebo Comparator<br>* Placebo (glucose powder)<br>|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Characterize Immunoglobulin fragment crystallization modifications | Use mass spectrometry and RNA sequencing from subject blood samples to characterize site specific glycosylation on Immunoglobulin A, Immunoglobulin M, and Immunoglobulin G. | 4 Weeks | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Assess for potential adverse events | Monitor symptoms, severity, and duration of adverse events. | 2 months after first dose | 
| Quantification of enzymatic modifications measured in grams by spectrometry | The investigators will measure enzymatic changes on glycoproteins in the immune system by spectrometry, i.e., these changes will be measured in grams. Spectometry will allow the investigators to quantify these changes in small detail, and therefore give a better idea of how supplementation can change enzymatic process that then affect the structure, and mass, of glycoproteins. | 6 months | 
| Flow cytometry to determine the effect of dietary supplementation on the immune system | Perform flow cytometry on subject blood samples to determine the effect of dietary supplement on immune system. For example, the investigators will note changes to immune cell pro-inflammatory and anti-inflammatory cytokine clusters as translated proteins (measured as height and area of cytokine intensity). | 6 months | 
| Transcriptome analysis to determine the effect of dietary supplementation on the immune system | Perform transcriptome analysis on subject blood samples to determine the effect of dietary supplement on immune system. For example, the investigators will note changes to immune cell pro-inflammatory and anti-inflammatory cytokine clusters as RNA (measure on a logarithmic scale per number of reads) to determine the effects of supplementation on both gene transcription and translation. | 6 months | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
dietary supplement