Deciphering the Role of Dietary Fatty Acids on Extracellular Vesicles-mediated Intercellular Communication

Study Overview
=================
Brief Summary
-----------------
Dietary interventions have been consistently proposed as a part of a comprehensive strategy to lower the incidence and severity of atherosclerosis and coronary vascular disease. Excessive comsumption of fats enriched in saturated fatty acids (SFAs) is associated with an increased risk of atherosclerosis and other cardiovascular diseases (CVD). In contrasts, replacement of SFAs with monounsaturated fatty acids (MUFAs) and omega-3 long chain polyunsaturated fatty acids (ω3-LCPUFAs) has been reported to be inversely associated with risk of atherosclerosis. This is partly due to the ability of MUFAs (and ω3-LCPUFAs) to modulate lipoprotein composition, oxidation state, and consequently their functionality, among others. While most of the nutritional studies have focused on elucidating the mechanisms by which dietary fats affect lipoprotein particles, little or nothing is known about the regulatory effect of dietary fatty acids on extracellular vesicles (EVs). EVs are small phospholipid particles that convey molecular bioactive cargoes and play essential roles in intercellular communication and, hence, a multifaceted role in health and disease. For the first time, the purpose of this project is to establish whether the type of major fatty acids present on a diet (SFAs, MUFAs, or ω3-LCPUFAs) may alter the structure, cargo, and functionality of postprandial- and long-term-EVs. In the precision nutrition era, the investigators expect to offer a new insight on EVs and their relationship with dietary fatty acids through the following objectives: 1) To map changes in the lipidome, proteome, microtranscriptome, and functional properties of circulating EVs in healthy subjects and patients with metabolic syndrome (MetS) both at fasting and at postprandial state upon a challenge of a meal rich in SFAs, MUFAs, and ω3-LCPUFAs; 2) To analyse the contribution of postprandial triacylglyceride-rich lipoproteins (TRL) on EVs-mediated intercellular communication in a fatty acid-dependent manner; and 3) To determine the influence of diets rich in SFAs, MUFAs, and ω3-LCPUFAs on EVs in an animal model of atherosclerosis in the setting of MetS. Collectively, this project will provide fundamental insight into EV biology, and remarks the clinical and functional relevance and divergent consequences of dietary fatty acids in health and disease.

Official Title
-----------------
Deciphering the Role of Dietary Fatty Acids on Extracellular Vesicles-mediated Intercellular Communication and Their Implication in Atherosclerosis and Metabolic Syndrome: a Multi-omics Approach to Precision Nutrition

Conditions
-----------------
Obesity, Metabolic Syndrome, Metabolic Disorder, Inflammation, Immune System and Related Disorders

Intervention / Treatment
-----------------
* Dietary Supplement: Oral lipid emulsions


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: • Clinical diagnosis of metabolic syndrome Exclusion Criteria: Allergy to dairy products Allergy to fish oil Vegetarian Tobacco smoker Current or recent (<4 wk) use of fish oil supplements or more than four times fish/week Received innoculations within 2 mo of starting the study or planned to during the study Donated or intended to donate blood from 2 mo before the study till 2 mo after the study Unstable body weight (no weight gain/loss >3 kg)

Ages Eligible for Study
-----------------
Minimum Age: 35 Years
Maximum Age: 65 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: Crossover Assignment
Masking: Single


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: Metabolic Syndrome patients<br> | Dietary Supplement: Oral lipid emulsions<br>* The oral lipid emulsions will contain water, sucrose, emulsifier, flavouring, and the corresponding fat (50 g/m2 of body surface area): milk cream (SFAs) or refined olive oil (MUFAs) with or without a dose of omega-3 PUFA, which will consist of 920 mg of EPA and 760 mg of DHA (ω3-LCPUFAs).<br>|
| Experimental: Healthy patients<br> | Dietary Supplement: Oral lipid emulsions<br>* The oral lipid emulsions will contain water, sucrose, emulsifier, flavouring, and the corresponding fat (50 g/m2 of body surface area): milk cream (SFAs) or refined olive oil (MUFAs) with or without a dose of omega-3 PUFA, which will consist of 920 mg of EPA and 760 mg of DHA (ω3-LCPUFAs).<br>|

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

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Evolution of cytokines in postprandial state | Pro-inflammatory and anti-inflammatory cytokines, including NFα, IL-1β, IL-6, IL-8, IL-10, ICAM-1, MCP-1, leptin, and adiponectin, in plasma will be measured using appropriate methods (EIA, ELISA, and/or Bioplex multiplex system) (mg/dl). | Up to 6 hours | 
| Evolution of inflammatory markers in postprandial state. [Time Frame: Up to 6 hours] | The acute phase protein (hsCRP), PAI-1, fibrinogen, transferrin, albumin, and myeloperoxidase (MPO) will be measured using appropriate methods (EIA, ELISA, and/or Bioplex multiplex system) (mg/dl). | Up to 6 hours | 
| Effect of EVs on gene expression in PBMCs | PBMCs will be isolated from the subjects' peripheral blood and treated with autologous circulating EVs for different times. | Up to 6 hours | 
| EV proteome | The quantification of exosome-derived proteins will be performed by nLC-MS/MS | Up to 6 hours | 
| EV lipidome | The analysis of intact lipids derived from exosomes will be performed by LC-MS | Up to 6 hours | 
| Ev microtranscriptome | Enriched RNA and miRNA derived from exosomes will be determined by Next Generation Sequencing (NGS) | Up to 6 hours |