Clinical Trial Details
— Status: Completed
Administrative data
| NCT number |
NCT05492084 |
| Other study ID # |
L-001 |
| Secondary ID |
|
| Status |
Completed |
| Phase |
|
| First received |
|
| Last updated |
|
| Start date |
January 1, 2019 |
| Est. completion date |
January 31, 2019 |
Study information
| Verified date |
August 2022 |
| Source |
Novosibirsk State Medical University |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Observational
|
Clinical Trial Summary
Вackground. Progressive atherosclerosis is accompanied by unfavorable clinical outcomes,
study and understanding of this process, creation of risk assessment method is necessary for
individualization of approaches to treatment and prevention of this condition.
Purpose of the study. Creation of a mathematical model to assess the risk of accelerated
atherosclerosis development, using methods of factor and correlation analysis.
Patient Characteristics and Study Methods. A retrospective cohort study included 202 patients
with coronary heart disease. Group 1 included patients who had had myocardial infarction or
unstable angina, emergency arterial stenting, stroke, peripheral artery thrombosis, critical
ischemia, and lower extremity amputation within 2 years before study inclusion. Patients in
the comparison group did not have these events. The influence of each of the studied
parameters on the probability of fast progressing atherosclerosis was determined by factor
and correlation analysis. The prospective part of the study will include follow-up of
patients from both groups for 12 months. Annual "endpoints": fatal outcome, unscheduled
coronary revascularization, nonfatal myocardial infarction and stroke, hospitalization due to
unstable angina pectoris, stent thrombosis, stenting/plasty of lower limb arteries.
Description:
Study design: open continuous retrospective and prospective study by parallel group method.
Three control points were defined in the dissertation work: the first point was
retrospective, 2 years before the study; the second was the day of inclusion in the study;
and the third was 12 months after inclusion.
The retrospective point covered 2 years from the date of patient inclusion in the study
(period from January 2019 to January 2021) with determination of a large spectrum of
parameters of clinical, instrumental, laboratory nature potentially influencing the
appearance and progression of atherosclerosis (according to the literature analysis). The
mentioned examinations were performed and evaluated at two points: the first one - at the
time of the first preceding coronary event during the two-year retrospective period, the
second one - at the time of the patient's inclusion in the present study. 202 patients were
divided into two groups.
Study Methods. At the moment of inclusion (1 point) in the study all patients underwent
general clinical examination with assessment of complaints and clinical status, laboratory
examination with general and biochemical blood tests, including special biochemical
examination of blood serum to determine the level of high-sensitivity C-reactive protein
(CRP, mg/L) and lipid spectrum, including total cholesterol (GC, mmol/L), low-density
lipoproteins (LDL, mmol/L), high-density lipoproteins (HDL, mmol/L), triglycerides (TG,
mmol/L); urinalysis and instrumental examinations: electrocardiography (ECG),
echocardiography (Echo-CG), R-graphy of chest organs, selective coronary angiography In
addition to standard research methods, all patients will be determined molecular genetic
parameters. Standard methods of investigation will be repeated after one year (2 points).
Preparation of DNA preparations. DNA extraction from blood was performed by phenol-chloroform
extraction. Five to six volumes of buffer A (10 mM Tris-HCl, pH=7.5; 10 mM NaCl; 3 mM MgCl2)
were added to 1 volume of blood sample and clots were rubbed in a homogenizer. After
centrifugation at 2500g for 15 min, the precipitates were washed three times with buffer A
and resuspended in 1 ml of buffer B (10 mM EDTA; 100 mM NaCl; 50 mM Tris-HCl, pH=8.5). After
adding SDS to 0.5% and proteinase E to 200 μg/mL, the mixture was incubated for 12 hours at
56°C. Deproteinization was performed sequentially with phenol-chloroform mixture (1:1),
water-saturated phenol, phenol-chloroform mixture (1:1), and chloroform. DNA was precipitated
by adding NaCl solution to 1 M and 1 V isopropyl alcohol. After that, the solution was cooled
for 1 h at -20 °C. The precipitate obtained by centrifugation on an Eppendorf microcentrifuge
at 12000g for 15 min was washed three times with 75% ethanol followed by centrifugation for 5
min. 12000g and, after drying at 56 °C, dissolved in deinanilized water to a DNA
concentration of 0.5 µg/μl.Genotyping of polymorphisms was performed using real-time PCR
according to the manufacturer's protocol (TaqMan probes, Thermo Fisher Scientific, USA) on a
StepOnePlus instrument. They were selected according to the results of international full
genome-wide association studies (GWAS), which confirmed the association of these SNPs with
coronary heart disease (CHD), an inflammatory process that plays a significant role in the
progression of atherosclerosis.
Methods of statistical analysis. SPSS 22.0 software package will be used for statistical
analysis of molecular-genetic data. The first stage will be to determine the frequencies of
genotypes and alleles of the studied SNPs in the group of myocardial infarction patients with
elevated values of cardiospecific markers and the comparison group, where cardiospecific
markers will be within normal limits; then we will assess compliance of genotype frequencies
with Hardy-Weinberg equilibrium in the control group (using the chi-square criterion). There
will be a comparison of the levels of indicators such as age; cre-atinine level; diagnostic
level of biomarkers of myocardial necrosis: MB creatine kinase and troponin, fatty acid
binding protein (FABP); body mass index; waist circumference; serum C-peptide concentration;
glycemia levels at admission and discharge; blood hemoglobin levels at admission and
discharge; Systolic and diastolic blood pressure at admission; heart rate at admission; left
ventricular ejection fraction value by echocardiography; serum HCMP concentration;
lipidogram: serum concentration of low-density lipoproteins (LDL), high-density lipoproteins
(HDL), total cholesterol, triglycerides; height, weight, body mass index, in carriers of
different genotypes were performed after checking normality of distribution of these features
by Kolmogorov-Smirnov test. If the traits would meet the criteria of normal distribution, a
single-factor analysis of variance would be used. Significance of differences between two
genotypic classes will be additionally checked using t-test for two independent samples. If a
trait under study does not meet the criteria of normal distribution, comparison of the level
of this trait in carriers of different genotypes will be performed using the Kruskal-Wallis
test; the reliability of differences between the two genotypic classes will be additionally
checked using the Mann-Whitney test for two independent samples. The association of SNPs with
risk factors related to categorical variables will be tested with conjugation tables using
Pearson chi-square test. In the case of four-field tables, comparison of samples by genotype
and allele frequencies will use Fisher's exact two-sided criterion. The relative risk (OR -
odds ratio) of disease outcome/risk factor presence for a particular allele or genotype will
be calculated as odds ratio.
Influence of clinical, demographic, functional, metabolic, inflammatory parameters and
myocardial necrosis markers on long-term prognosis will be assessed by odds ratio (0.95
confidence interval).
