Atherosclerosis, Cardiovascular Diseases, Low-density Lipoprotein Lipid Composition, Low-density Lipoproteins Aggregation Susceptibility
Conditions
Keywords
Atherosclerosis, E-EPA, LDL aggregation susceptibility, Lipidomics, Proteoglycan binding, Icosapent ethyl, Fatty acid, Metabolism, Omega-3
Brief summary
40-70 healthy volunteers of ages 18 to 65 participate in a E-EPA-diet where 3,9 grams of E-EPA is added to their normal diet and lifestyles for a month. Blood samples will be collected before the study and at weeks 1 and 4 and also, two weeks after finishing the diet. Main study focuses are LDL aggregation susceptibility, lipid composition and proteoglycan binding affinity. In addition, important plasma lipid metabolism enzymes and lipid mediated resolvins are measured as well as several baseline characteristics.
Detailed description
Four grams of daily E-EPA was found to significantly decrease atherosclerotic cardiovascular diseases in the REDUCE-IT study. In the upcoming study the effect of E-EPA to low-density lipoprotein (LDL) aggregation susceptibility is measured using dynamic light scattering technique to continuously measure particles size while inducing aggregation. LDL Lipid composition is analyzed using electrospray mass spectrometer optimized for lipid measurements. Previously is reported that the aggregation susceptibility is affected by the lipid composition which is modifiable (Ruuth et. al. Eur.H.Journal 2018). Common disease factors such as total cholesterol, LDL, HDL, triglycerides, ApoB-100, ApoA-I, Lp(a) and different modified LDL levels are measured. Also, activities of lipid metabolism enzymes like PON-1, LCAT, CETP, PLTP and lipid mediated resolvins are looked into. Objective is to identify changes and possible pathways that are altered by the increase of E-EPA and to use the data to possibly explain the health benefits of EPA. Update at study conclusion 17.6.2025. Covid-19 halted our study progression leading to a partial cohort (final n=38). Covid-19 restrictions also prevented us from measuring plasma lipid metabolism enzymes and lipid mediated resolvins. As a results we expanded the lipoprotein lipidomics analysis to consist all three lipoprotein fractions (VLDL, LDL, and HDL) using LC/MS. Finally a clinical cardiovascular risk score assessment was also performed (CERT2/Hertta-test).
Interventions
DIETARY_SUPPLEMENTEthyl-Eicosapentaenoic Acid (E-EPA)
3,9 grams of E-EPA (Icosapent ethyl) is added to participants' normal diet.
Sponsors
University of Helsinki
Wihuri Research Institute
Study design
Allocation
NA
Intervention model
SINGLE_GROUP
Primary purpose
BASIC_SCIENCE
Masking
NONE
Intervention model description
Baseline measurements are compared to different time points during the study.
Eligibility
Sex/Gender
ALL
Age
18 Years to 65 Years
Healthy volunteers
Yes
Inclusion criteria
* Healthy normolipidemic
Exclusion criteria
* Prescription of blood thinner medicine * Circulating Low-density lipoprotein \> 5mmol/l, Triglycerides \>3mmol/l * Chronic use of pain medication * Fish allergy * Pregnancy * Breastfeeding
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| LDL Aggregation Susceptibility | 28 days | LDL aggregation susceptibility was induced in vitro by sphingomyelinase and measured using dynamic light scattering. Time-size curves were generated, and the inflection point (EC50)-the midpoint of the most rapid aggregation-was determined by nonlinear regression with a modified Hill equation. A longer time to reach EC50 indicates lower aggregation susceptibility, and thus a lower risk of future cardiovascular events. |
| Total Blood Triglycerides | 28 days | Percentage change in blood triglycerides after IPE-supplementation (day 28) compared to the baseline (day 0). Percentage change was calculated as follows: \[(day 28 - day 0) / day 0\] x 100. |
| EPA Incorporation Into LDL | 28 days | Total concentration of eicosapentaenoic acid in LDL lipoprotein fraction at baseline (day 0), and after IPE-supplementation (day 28). |
Other
| Measure | Time frame | Description |
|---|---|---|
| Lipoprotein Retention | 28 days | The binding of lipoproteins to aortic proteoglycans, measured ex vivo. At the end of the assay lipoprotein-associated bound cholesterol is measured in each well and compared to control wells to determine the binding probability of LDL particles to aortic proteoglycans. |
| Coronary Event Risk Test 2 | 28 days | A clinical risk rest used to assess 10-year Coronary event risk. Based on plasma ceramides and phospholipids. Lower risk score indicates smaller future risk of coronary events. The four risk categories are: 0-3 (Low risk), 4-6 (Moderate risk), 7-8 (High risk), and 9-12 (Very high risk). |
Countries
Finland
Participant flow
Recruitment details
Recruitment took place in autumn/winter of 2019/2020 via in-house emailing lists. Participant suitability was assessed prior to the beginning of the supplementation. Healthy normolipidemic individuals aged 18-65 were enrolled.
Participants by arm
| Arm | Count |
|---|---|
| IPE-supplementation Group All the study participants will receive the same treatment. 3.9g of IPE (Ethyl-EPA) in capsules, which also include 75µg of D3-vitamin, daily for 28 days. | 38 |
| Total | 38 |
Withdrawals & dropouts
| Period | Reason | FG000 |
|---|---|---|
| Overall Study | Covid-19 restrictions | 27 |
| Overall Study | Withdrawal by Subject | 3 |
Baseline characteristics
| Characteristic | IPE-supplementation Group |
|---|---|
| Age, Categorical <=18 years | 0 Participants |
| Age, Categorical >=65 years | 0 Participants |
| Age, Categorical Between 18 and 65 years | 38 Participants |
| Blood triglycerides | 0.9 mmol/L STANDARD_DEVIATION 0.4 |
| Body Mass Index (BMI) | 23.5 kg/m2 STANDARD_DEVIATION 2.5 |
| LDL-cholesterol | 1.7 mmol/L STANDARD_DEVIATION 0.4 |
| Race (NIH/OMB) American Indian or Alaska Native | 0 Participants |
| Race (NIH/OMB) Asian | 0 Participants |
| Race (NIH/OMB) Black or African American | 0 Participants |
| Race (NIH/OMB) More than one race | 0 Participants |
| Race (NIH/OMB) Native Hawaiian or Other Pacific Islander | 0 Participants |
| Race (NIH/OMB) Unknown or Not Reported | 0 Participants |
| Race (NIH/OMB) White | 38 Participants |
| Region of Enrollment Finland | 38 Participants |
| Sex: Female, Male Female | 27 Participants |
| Sex: Female, Male Male | 11 Participants |
Adverse events
| Event type | EG000 affected / at risk |
|---|---|
| deaths Total, all-cause mortality | 0 / 38 |
| other Total, other adverse events | 0 / 38 |
| serious Total, serious adverse events | 0 / 38 |
Outcome results
Primary
EPA Incorporation Into LDL
Total concentration of eicosapentaenoic acid in LDL lipoprotein fraction at baseline (day 0), and after IPE-supplementation (day 28).
Time frame: 28 days
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| IPE-supplementation Group | EPA Incorporation Into LDL | Baseline | 0.075 mmol/L | Standard Deviation 0.094 |
| IPE-supplementation Group | EPA Incorporation Into LDL | After 28-day of IPE-supplementation | 0.274 mmol/L | Standard Deviation 0.118 |
Primary
LDL Aggregation Susceptibility
LDL aggregation susceptibility was induced in vitro by sphingomyelinase and measured using dynamic light scattering. Time-size curves were generated, and the inflection point (EC50)-the midpoint of the most rapid aggregation-was determined by nonlinear regression with a modified Hill equation. A longer time to reach EC50 indicates lower aggregation susceptibility, and thus a lower risk of future cardiovascular events.
Time frame: 28 days
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| IPE-supplementation Group | LDL Aggregation Susceptibility | Baseline | 111.3 Minutes | Standard Deviation 17.3 |
| IPE-supplementation Group | LDL Aggregation Susceptibility | After 28-day of IPE-supplementation | 113.1 Minutes | Standard Deviation 25.8 |
Primary
Total Blood Triglycerides
Percentage change in blood triglycerides after IPE-supplementation (day 28) compared to the baseline (day 0). Percentage change was calculated as follows: \[(day 28 - day 0) / day 0\] x 100.
Time frame: 28 days
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| IPE-supplementation Group | Total Blood Triglycerides | -14 percentage of baseline triglycerides | Standard Deviation 28 |
Other Pre-specified
Coronary Event Risk Test 2
A clinical risk rest used to assess 10-year Coronary event risk. Based on plasma ceramides and phospholipids. Lower risk score indicates smaller future risk of coronary events. The four risk categories are: 0-3 (Low risk), 4-6 (Moderate risk), 7-8 (High risk), and 9-12 (Very high risk).
Time frame: 28 days
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| IPE-supplementation Group | Coronary Event Risk Test 2 | Baseline | 3.4 Scores on a scale | Standard Deviation 2.6 |
| IPE-supplementation Group | Coronary Event Risk Test 2 | After 28-day of IPE-supplementation | 2.5 Scores on a scale | Standard Deviation 1.8 |
Other Pre-specified
Lipoprotein Retention
The binding of lipoproteins to aortic proteoglycans, measured ex vivo. At the end of the assay lipoprotein-associated bound cholesterol is measured in each well and compared to control wells to determine the binding probability of LDL particles to aortic proteoglycans.
Time frame: 28 days
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| IPE-supplementation Group | Lipoprotein Retention | Baseline | 4.995 Bound cholesterol nmol/well | Standard Deviation 0.041 |
| IPE-supplementation Group | Lipoprotein Retention | After 28-day of IPE-supplementation | 4.428 Bound cholesterol nmol/well | Standard Deviation 0.039 |