Search

Aim.To search for causal mutations in candidate genes responsible for the development of sudden cardiac death (SCD) in men who died under the age of 45.Materialandmethods.The SCD group (n=37) was formed using the criteria the World Health Organization and the European Society of Cardiology. Autopsy material was collected from men who died suddenly outside medical institutions and underwent forensic medical examination according to the standard protocol. Autopsy revealed no morphological changes that could explain sudden death. The mean age was 32,4±6,4 years. Genomic DNA was isolated from myocardial tissue using phenol-chloroform extraction. Clinical exome sequencing was performed. At first, we analyzed the results of sequencing of 24 genes, mutations in which lead to cardiovascular diseases associated with an increased risk of SCD:KCNQ1,KCNH2,SCN5A,AKAP9,ANK2,CACNA1C,CALM1,CALM2,CAV3,KCNE1,KNCJNE2,KCNE2,SCN4B,SNTA1,MYH2,APOB,KCNA5,TGFB3,NEB,PDX1,FLNC,PLEC,KCND3.Results.Of 37 samples, we revealed 13 probable pathogenic missense mutations in 9 samples (24,3%). Of 13 probable pathogenic variants, 5 were new.Conclusion.This pilot study provides following conclusions: it is necessary to continue molecular autopsy research in Russia; to increase the effectiveness of detecting causal mutations, it is necessary to reduce the age of patients with SCD included in the study; studying the families of deceased; cooperation of experienced specialists — forensic pathologist, laboratory geneticist, cardiologist.

Aim . To study the association of new polymorphisms with sudden cardiac death (SCD) in men. Material and methods . The SCD group (n=278) was formed using guidelines of the World Health Organization and the European Society of Cardiology. The autopsy material was taken from the men who died suddenly outside the prevention and treatment facilities who underwent medicolegal examination according to a standard protocol. The average age in the SCD group was 53,2±8,7 years. The control group (n=274) was selected by sex and age from the DNA bank formed during the international studies of MONICA and HAPIEE. The mean age of men in the control group was 53.±8,3 years. Genomic DNA was separated from myocardial tissue and venous blood by the method of phenol-chloroform extraction. Rs10503929, rs7121, rs6730157, rs11720524, rs7737692, rs7521023 polymorphisms were tested using real-time PCR. The insertion- deletion polymorphisms rs.0692285 and rs39.7 were genotyped by PCR with flanking primers followed by polyacrylamide gel electrophoresis. Statistical analysis was performed using the SPSS 13.0 software package. Results . By comparison the frequencies of rs7121 genotypes in the studied groups of Novosibirsk, an increase in the proportion of CC genotype carriers in the group with SCD (OR=.6, 95% CI 1,1-2,2; p=0,025) was found. The carriage of the rs.0692285 ID genotype reduces the risk of SCD (OR=0,6, 95% CI 0,4-0,9; p=0,0M). A carriage of the rs39.7 ID genotype increases the risk of SCD (OR=.5, 95% CI 1,1-2,2; p=0,020). Conclusion . The rs7121 of the GNAS gene, the rs.0692285 of the RYR2 gene, and the rs39.7 of the COL1A2 gene are associated with SCD. Rs10503929, rs6730157, rs11720524, rs7737692, rs7521023 polymorphisms do not associate with SCD in the men of Novosibirsk.

Genetic background of hypertension (AT) and blood pressure (BP) regulation is extensively investigated in genome-wide association studies (GWAS). The findings from recent GWAS require replication in independent samples. Aim . To investigate the association between BP and AT in Russian population and several single nucleotide polymorphisms identified in GWAS. Material and methods . In the frame of “case-control” design we recruited subjects with AT diagnosed at age below 50 according to the criteria of BP >140/90 mm Hg and/or receiving antihypertensive therapy, and subjects with normotension according to 2 examinations from population sampling, Novosibirsk, totally included 514, men/women aged 45-69 years). From published GWAS we selected 24 genetic markers related to hypertension, 8 markers were included for present analysis (rs 13082711, rs1173771, rs13107325, rs3918226, rs1799945, rs805303, rs1458038, rs932764). Standard epidemiological methods were used (BP measurement, anthropometry, medical history of AT and treatment, risk factors of AT, socio-demographic parameters). Single nucleotide polymorphisms (SNPs) were tested using real time PCR. Results . In studied sample we replicated the association between rs3918226 (promoter region of gene of endothelial NO synthase; eNOS) and systolic BP (T-allele carriers had 9 mm Hg higher systolic BP than CC carriers in men, p=0,049 independent of age). New association was found between rs 932764 (gene of phospholipase-c-epsilon-1 isoform, PLCE1) and AT (heterozygotes genotype AG was protective in men, p=0,017 independent of age and body mass). The association between rs13107325 (gene of soluble carrier family 39/zinc transporter/member 8, SLC39A8) and systolic BP was confirmed (in men, С-allele carriers had higher systolic BP values then TT carriers, p=0,044, multivariable adjusted). Conclusion . In analysis of relationship between phenotypes of BP and AT and 8 genetic markers of AT in Russian population sample we replicated two known associations, revealed new association and identified new data on modulating effect of sex and body mass. These replications in newly studied Siberian population, different from early studied populations by risk factors profile, climate, geographic and other parameters, support the involvement of identified or close loci in potential mechanisms of AT susceptibility.

Aim. To evaluate the utilization of complex significance evaluation in Russian population, of genetic markers of myocardial infarction identified in entire-genomic associative studies. Material and methods . Group of MI patients (n=200) and control group (n=420) were created based on populational selection of 45-69 year old citizens of Novosibirsk (9400 persons), collected during the international project HAPIEE; myocardial infarction patients, admitted to intensive care unit of the Hospital № 1 (160 persons); sudden cardiac death victims, post forensic investigation (n=285). Longevity group was collected during home visits and in the nursing houses (n=85). Genomic DNA was extracted from venous blood and myocardial tissue by phenolchlorophorm method. Genes polymorphism was tested with PCR real-time according to the protocol by equipment developer (TaqMan, Applied Biosystems, USA), device ABI 7900HT. For the study, the following MNP were selected: rs499818, rs619203, rs10757278 and rs1333049 (chr. 9), rs1376251, rs2549513, rs4804611, rs17465637. Results. Mononucleotide polymorphisms, that did not show differences at pretest stage, showed significant distinctions in the studied population. Two MNP were closely linked: rs10757278 and rs1333049 (chr. 9) so for further assessment the rs1333049 was retained. Among other seven, only five MNP showed relation with MI of various strength. Some, like the rs1333049 showed association with MI in all groups and subgroups, while the others — only in separate groups, with restriction by sex and gender. The rs2549513 and rs17465637 showed to be not associated with CHD at all, though rs2549513 showed association with lipid and glucose metabolism, as with the severity of coronary atherosclerosis and adverse prognosis for 1 year post-MI. The rs17465637 was just poorly associated with BMI. Conclusion. Complex approach to assessment of polymorphism role is quite demanding, but provides with a high grade of insurance in the significance of the results obtained, of non-random character of the results, and makes it to choose the most credible, that have showed association not only with the main pathological phenotype, but with its risk factors.

Objective: Investigate the association of rs17465637 gene MIAF3 (1q41), rs1376251 gene TAS2R50 (12p13), rs4804611 gene ZNF627 (19p13), rs619203 gene ROS1 (6q22), rs1333049 (9p21), rs10757278 (9p21), rs2549513 (16q23), rs499818 (6p24) associated with myocardial infarction available from the international genome-wide studies with sudden cardiac death (SCD) in a case–control study. Methods: A sample of SCD cases (n = 285) was formed using the WHO criteria; the control sample (n = 421) was selected according to sex and age. DNA was isolated by phenol–chloroform extraction from the myocardial tissue of SCD cases and blood of control cases. The groups were genotyped for the selected SNPs by real-time PCR using TaqMan probes (Applied Biosystems, United States). Results: No statistically significant differences in the genotype and allelic frequencies of studied single nucleotide polymorphisms between sudden cardiac death cases and control were detectable in general group. By separating the groups of sex and age differences in the genotype frequencies of rs1333049, rs10757278 and rs499818 are statistical significance. Genotypes CC of rs1333049 and GG of rs10757278 are associated with an increased sudden cardiac death risk in men (p = 0.019, OR = 1.7, 95% CI 1.1–2.8; p = 0.011, OR = 1.8, 95% CI 1.2–2.8, respectively). Genotype AG of rs499818 is associated with an increased sudden cardiac death risk in the women over 50 years old (p = 0.009, OR = 2.4, 95% CI 1.3–4.6). Conclusion: Polymorphisms rs1333049 and rs10757278 are associated with SCD in men and rs499818 in the women aged over 50 years.

Aim. To search and study the association of some candidate genes polymorphysms of various cardiovascular diseases and sudden cardiac death in men. Material and methods. The sudden cardiac death group (SCD) is collected by the WHO criteria with suddenly died men underwent court-medicine expertise (n=274). Control group was matched by the age and gender from the DNA bank of HAPIEE and MONICA trials. Genotyping of groups was done according to polymorhysms SCN5A, rs187238 gene IL-18, rs1799864 gene CCR2, rs3864180 gene GPC5, rs1799983 gene eNOS, rs2228314 gene SREBF-2, rs1800588 gene HL, rs10757278, rs1333049 with methods of PDRF and realtime PCR. Results. No significant differences found between the SCD group and control group by the prevalence of genotypes SCN5A, rs187238 gene IL-18, rs3864180 gene GPC5, rs1799983 gene eNOS, rs1800588 gene HL. In SCD group the decrease of homozygotes by CC polymorphism rs2228314 found in gene SREBF-2 and the decrease of heterozygotes GC comparing to control group (HR=4,074, 95% CI 1,843-9,002, р=0,0002; HR=0,442, 95% CI 0,302-0,647, р=0,0001, reap.). In SCD group carriers of GG genotype polymorphism rs10757278 and CC poly morphism rs1333049 are significantly more prevalent than in control group (HR=1,814, 95% CI 1,159-2,839, р=0,011; HR=1,744, 95% CI 1,104-2,754, р=0,019, resp.). The part of GA carriers of polymorphism rs1799864 gene CCR2 is more prevalent in SCD group comparing to control (HR=1,558, 95% CI 1,051-2,308, р=0,029). Conclusion. Polymorphisms rs10757278, rs1333049, rs2228314 gene SREBF-2, rs1799864 gene CCR2 are associated with sudden cardiac death in men.

Aim. The study of mononucleotide polymorphisms associations rs20455 gene KIF6, rs7439293 gene PALLD, rs2298566 gene SNX19, rs3900940 gene MYH15, rs1010 gene VAMP8, included into the Riskometer of ischemic heart disease of the Celera Corporation (USA), in sudden cardiac death. Material and methods. To the study the group of persons included, died from sudden cardiac death, according to the WHO criteria (n=352, mean age — 53,3±8,9 y.), control group from matched by age and gender from DNA bank of HAPIEE, MONICA studies (n=381, mean age — 53,1±8,3 y.), adolescents group (n=296, mean age — 15,6±0,9 y.). DNA extracted via the phenol-chloroform extraction from myocardium of the dyed due to sudden cardiac death, and from venous blood of controls. Genotyping performed with Real-time PCR method using the Taq-Mancatheters technology (Applied Biosystems). Results. The prevalence of alleles rs7439293 gene PALLD, rs2298566 gene SNX19, rs3900940 gene MYH15 did not statistically significantly differ among the groups. In those died suddenly older than 50 years, there was significant decrease of carriers of GG polymorphism rs20455 gene KIF6 (9,0%) comparing to the controls (17,8%) (р=0,009, OR=0,456, 95% CI 0,256-0,810). In men older 50, died suddenly, there was significant decrease of the carriers of genotype СС (8,0%) (р=0,002, OR=0,328, 95% CI 0,159-0,678) and increase of СТ genotype (49,6%) (р=0,025, OR=1,729, 95% CI 1,084-2,758) polymorphism rs1010 gene VAMP8 comparing to the controls (21,0%, 36,3%, respectively). Conclusion. Polymorphisms rs20455 gene KIF6 and rs1010 gene VAMP8 are associated with sudden cardiac death.

Aim. To check whether the mononucleotide polymorphisms (MNP), identified in recent studies do match a risk marker criteria for arterial hypertension (AH) in Siberian population.Material and methods. The group of subjects with AH and control group (relation 2:1) were formed from the population selection of 45-69 years old citizens of Novosibirsk (9400 subjects), which had been collected during the work in HAPIEE project. Totally 514 subjects included, of those controls – 168 (127 men and 41 women) with BP not higher than “normal” by the data of 2 and more examinations during the last years, with interval not more than 6 months. Group with AH consists of 346 subjects (206 men and 140 women) with the diagnosis of AH. Genomic DNA was extracted from venous blood by the method of phenol-chloroform extraction. Gene polymorphism was tested by PCR in real time according to the protocol of equipment manufacturer (zonds TagMan, Applied Biosystems, USA) on the device ABI 7900HT. These MNP were included into the study: rs699 gene AGT, rs5068 gene NPPB, rs17367504 gene MTHFR, rs2681492 gene ATP2B1, rs4343 gene АСЕ, rs1801253 gene ADRB1, rs11240692 gene REN, rs2846679 gene KCNJ1, rs239345 gene SCNN1B, rs1799983 gene NOS3.Results. The risk relation to develop AH in men – carriers of GG rs699 gene AGT was1,95 (95% CI 1,08-3,53; p=0,003) comparing to the carriers of two other genotypes. Carriage of AA genotype rs699, opposite, is probably protective factor for AH development in men (HR 0,47; 95% CI 0,27-0,81; p=0,007).Conclusion. For two from ten MNP studied the association with AH was confirmed in men: rs699 gene AGT and rs5068 gene NPPB.

Aim. To evaluate the relationship of 8 mononucleotide polymorphisms (MNP) with negative longterm prognosis of the acute coronary syndrome. Material and methods. Totally 280 patients with acute coronary syndrome included (ACS), who during 1 year follow up were underwent the solid endpoints evaluation (cardiovascular death, hospitalization for recurrent ischemic event and repeated revascularization). Results. In the analysis of rs4804611 in group with negative outcomes there is significantly more prevalent genotype GG (9,1% vs. 1,8%), chances relation 5,57 (95% CI 1,35-22,96); credibility of the differences kept in men (p=0,021). In the analysis rs2549513 in the group with positive prognosis the AA genotype was more prevalent (70,6% vs. 53%), hazard ratio 0,47 (95% CI 0,26-0,85), and in the group with negative prognosis genotype AC, chances relation 2,32 (95% CI 1,28-4,20; р=0,007). While separated by gender credible differences were shown only in men (p=0,008). Conclusion. Evaluation of genetic markers rs4804611 and rs2549513 can significantly increase prognostic value in riskometry of ACS outcomes.