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YI-805 SYNERGISTIC EFFECTS OF CHOLESTERYL ESTER TRANSFER PROTEIN, APOLIPOPROTEIN E, AND APOLIPOPROTEIN C3 GENE POLYMORPHISMS ON CORONARY ARTERY DISEASE RISK
214 YI-802
YI Young investigators MOLECULAR CHARACTERIZATION OF FAMILIAL HYPERCHOLESTEROLAEMIA IN PORTUGUESE PATIENTS
A.M. Medeiros, A.C. Alves, S. Silva, M. Bourbon on behalf of the Portuguese FH group. Unidade de Investigacao Cardiovascular, Instituto Nacional de Saude Dr. Ricardo Jorge, Lisboa, Portugal Background: Familial Hypercholesterolaemia (FH) is an autosomal dominant disorder caused by mutations in LDL receptor (LDLR) gene but defects in genes like APOB or PCSK9 also cause a similar phenotype. FH is characterized by defective clearance of plasma LDL that results in increased circulating LDL-cholesterol, leading to premature atherosclerosis and coronary heart disease (CHD). Portugal should have about 20000 cases of FH. Since 1999 the “Portuguese FH Study” has studied more than 500 individuals (www.spc.pt/hgs). Aims: The aim of this study is the identification of mutations in the LDLR, APOB and PCSK9 genes in patients with clinical diagnosis of FH. Methods: Blood samples were collected from 175 unrelated patients with clinical diagnosis of FH and 411 (affected and unaffected) relatives. LDLR gene was analysed using several molecular biology techniques such as PCR, DHPLC, direct sequencing and MLPA. One fragment corresponding to the exon 26 of the APOB gene and the 12 exons of the PCSK9 were analysed by PCR and direct sequencing. Results and Conclusion: Fifty eight different mutations were detected in 212 individuals, 34 previously described and 24 novel mutations possibly affecting the function of the LDLR. Two of these patients were homozygous and 2 patients were compound heterozygous. Additionally, 6 subjects carried the APOB3500 mutation and 3 had a PCSK9 mutation. These results confirm that LDLR gene has a wide spectrum of sequence variations in our population. The molecular identification of FH patients is important for these patients to receive the correct treatment to prevent premature CHD. YI-803
POLYMORPHISM IN THE CYP26B1 GENE AND ASSOCIATION WITH THE SUSCEPTIBILITY OF MYOCARDIAL INFARCTION
O. Krivospitskaya 1 , K. Jatta 1 , L. Johansson 3 , J.H. Jansson 3 , A. Hurtig-Wennlof 1 , P. Olofsson 2 , A. Sirsjo 1 . 1 Division of Biomedicine, Department of Clinical Medicine, University of Orebro, Orebro, Sweden; 2 Center for Molecular Medicine, Cardiovascular Research Unit, Karolinska Institute, Stockholm, Sweden; 3 Department of Internal Medicine, Skellefta Hospital and Umea University Hospital, Umea, Sweden Background and Aims: Cardiovascular diseases have a complex etiology. A large number of different processes are involved, such as cell migration, proliferation, differentiation, immune response and coagulation/fibrinolysis. Retinoids regulate a vast number of processes involved in it. CYP26B1 is one enzyme of P450-related retinoic acid hydroxylases that catabolise atRA to inactive metabolites and are important regulators of cellular retinoid levels. In this study we investigated rs2241057 SNP of CYP26B1 and the association with the risk of MI. Material and Methods: We genotyped 393 male patients with MI and 735 healthy male controls and 152 female patients and 273 healthy female controls by allelic discrimination using TagMan. Cross-sectional study of allelic frequencies was performed using Chi-Square Tests. Results: T-allel of rs2241057 SNP occurs more frequently in the cases of myocardial infarction compared to healthy controls in male population, C-allel is significantly less frequent in male patients compared to healthy controls (Pearson Chi-Square = 0,063, Fisher‘s Exact Test = 0,035). Multiple logistic analysis showed that C-allel was a protective factor for males with an odds ratio of 0,059. No difference was found between cases and controls among females. Conclusions: We found a strong tendency between frequency of occurrence of rs2241057 SNP C-allel and diminished number of cases of MI in male population, but not in the female population. Further research is needed to evaluate the role of CYP26B1 in cardiovascular disease. YI-804
NEW MARKERS OF OXIDATIVE STRESS AND ENDOTHELIAL DYSFUNCTION IN MYOCARDIAL INFARCTION
Y.I. Ragino, V.A. Baum, Y.V. Polonskaya, E.V. Sadovski. Institute of Internal Medicine SB RAMS, Novosibirsk, Russia Aim of the study was to investigate the relationships between oxidized
fibrinogen, oxidized apolipoproteins of low density lipoproteins (apo-LDL) and activity of plasma lipid peroxidation (LPO) process, endothelial dysfunction parameters and disturbances of hemostasis system in myocardial infarction (MI). 112 men 39-65 years old were included into the study: 43 group-1 men with MI in anamnesis last year, 19 group-2 men with subacute MI, 50 group-3 men without CHD. New methods for evaluation of oxidized fibrinogen and apo-LDL (patents of Russia) were worked out. Parameters of LPO process of plasma and LDL were determined as TBAreactive substances. Oxidative resistance of LDL was studied during LDL incubation with copper ions by spectrofluorimetrical method. Parameters of blood hemostasis system (antithrombine III, AT-III, thrombin inactivation index, TII, activity of XIIa-dependent fibrinolysis and other) and plasma level of NO metabolites were determined. Plasma levels of vonWillebrand factor and of endothelin-1 were evaluated by ELISA method. The extent of oxidized fibrinogen was 4 times higher (p<0,001) in group-2 men versus group-3 men. The level of oxidized apo-LDL was higher (p<0,01) in group-1 and group-2 men versus group-3 men. Independent association of oxidative modified fibrinogen with MI was revealed (p<0,001). Group-2 men had increased AT-III, TII and decreased activity of XIIa-dependent fibrinolysis as compared with group-3 men. Strong positive correlations between parameters of plasma LPO process and endothelial dysfunction and oxidized fibrinogen level were found (p<0,01). It is possible that oxidized fibrinogen may serve as biomarkers for MI. YI-805
SYNERGISTIC EFFECTS OF CHOLESTERYL ESTER TRANSFER PROTEIN, APOLIPOPROTEIN E, AND APOLIPOPROTEIN C3 GENE POLYMORPHISMS ON CORONARY ARTERY DISEASE RISK
A. Muendlein 1 , C.H. Saely 1,2 , S. Aczel 1,2 , T. Marte 1,2 , F. Schmid 1 , L. Koch 1 , P. Rein 1,2 , P. Langer 1 , H. Drexel 1,2,3 . 1 VIVIT Institute, Feldkich, Austria; 2 Academic Teaching Hospital Feldkirch, Feldkich, Austria; 3 University for Human Sciences, Triesen, Liechtenstein Background and aims: We aimed at investigating potential synergistic effects of cholesteryl ester transfer protein (CETP), apolipoprotein E (apoE), and apolipoprotein C3 (apoC3) gene variants on the risk of coronary artery disease (CAD). Methods: We analysed the CETP TaqIB, the APOE E3/E4/E2 and the APOC3 –482 C>T polymorphisms in 557 patients undergoing coronary angiography. Significant CAD was defined as the presence of coronary stenoses ≥50%. Results: From our patients, 129 carried the APOE E4 allele, 278 the APOC3 -482T allele and 196 the CETP B1B1 genotype. After multivariate adjustment, presence of the APOE E4 allele proved significantly associated with the presence of significant CAD (odds ratio 1.589 [95% CI 1.013-2.491]; p = 0.044); odds ratios for the APOC3 -482T allele and the CETP B1B1 genotype were 1.170 [0.813-1.686]; p = 0.398 and 1.453 [0.985-2.142]; p = 0.059, respectively. The risk of significant CAD was strongly increased in patients with more than one of the analysed variants: adjusted odds ratios were 1.831 [1.113-3.013]; p = 0.017 for patients with both the APOE E4 allele and the CETP B1B1 genotype, 1.983 [1.017-3.868]; p = 0.045 for patients with both the APOE E4 and the APOC3 -482T alleles, 2.498 [1.474-4.231]; p = 0.001 for patients with both the CETP B1B1 genotype and the APOC3 -482T allele, and 4.491 [1.222-16.499]; p = 0.028 for patients with all three variants. Conclusions: There are strong synergistic effects of the CETP TaqIB, APOE E3/E4/E2 and APOC3 –482 C>T polymorphisms on the risk of angiographically diagnosed CAD. YI-806
PCSK9 GENE ANALYSIS IN PORTUGUESE PATIENTS WITH FAMILIAL HYPERCHOLESTEROLEMIA
A.C. Alves 1 , Q. Rato 2 , A. Furtado 3 , M. Bourbon For The Portuguese Fam 1 . 1 Unidade de Investigacao Cardiovascular, Instituto Nacional de Saude Dr. Ricardo Jorge, Lisboa, Portugal; 2 Serv. de Cardiologia, Centro Hospitalar de Setubal, E.P.E., Hospital de S. Bernardo, Setubal, Portugal; 3 Serv. de Medicina Interna, Hosp. Pedro Hispano, Matosinhos, Portugal Background: Familial hypercholesterolemia (FH) is an autosomal dominant disorder associated with high risk of coronary heart disease. FH usually results from inherited defects in the Low Density Lipoprotein receptor gene (LDLR) and is characterised by increased circulating LDL cholesterol that leads to lipid accumulation in arteries and tendons (xanthomas), causing premature arteriosclerosis and coronary heart disease. Mutations
76th Congress of the European Atherosclerosis Society, June 10–13, 2007, Helsinki, Finland
YI Young investigators MOLECULAR CHARACTERIZATION OF FAMILIAL HYPERCHOLESTEROLAEMIA IN PORTUGUESE PATIENTS
A.M. Medeiros, A.C. Alves, S. Silva, M. Bourbon on behalf of the Portuguese FH group. Unidade de Investigacao Cardiovascular, Instituto Nacional de Saude Dr. Ricardo Jorge, Lisboa, Portugal Background: Familial Hypercholesterolaemia (FH) is an autosomal dominant disorder caused by mutations in LDL receptor (LDLR) gene but defects in genes like APOB or PCSK9 also cause a similar phenotype. FH is characterized by defective clearance of plasma LDL that results in increased circulating LDL-cholesterol, leading to premature atherosclerosis and coronary heart disease (CHD). Portugal should have about 20000 cases of FH. Since 1999 the “Portuguese FH Study” has studied more than 500 individuals (www.spc.pt/hgs). Aims: The aim of this study is the identification of mutations in the LDLR, APOB and PCSK9 genes in patients with clinical diagnosis of FH. Methods: Blood samples were collected from 175 unrelated patients with clinical diagnosis of FH and 411 (affected and unaffected) relatives. LDLR gene was analysed using several molecular biology techniques such as PCR, DHPLC, direct sequencing and MLPA. One fragment corresponding to the exon 26 of the APOB gene and the 12 exons of the PCSK9 were analysed by PCR and direct sequencing. Results and Conclusion: Fifty eight different mutations were detected in 212 individuals, 34 previously described and 24 novel mutations possibly affecting the function of the LDLR. Two of these patients were homozygous and 2 patients were compound heterozygous. Additionally, 6 subjects carried the APOB3500 mutation and 3 had a PCSK9 mutation. These results confirm that LDLR gene has a wide spectrum of sequence variations in our population. The molecular identification of FH patients is important for these patients to receive the correct treatment to prevent premature CHD. YI-803
POLYMORPHISM IN THE CYP26B1 GENE AND ASSOCIATION WITH THE SUSCEPTIBILITY OF MYOCARDIAL INFARCTION
O. Krivospitskaya 1 , K. Jatta 1 , L. Johansson 3 , J.H. Jansson 3 , A. Hurtig-Wennlof 1 , P. Olofsson 2 , A. Sirsjo 1 . 1 Division of Biomedicine, Department of Clinical Medicine, University of Orebro, Orebro, Sweden; 2 Center for Molecular Medicine, Cardiovascular Research Unit, Karolinska Institute, Stockholm, Sweden; 3 Department of Internal Medicine, Skellefta Hospital and Umea University Hospital, Umea, Sweden Background and Aims: Cardiovascular diseases have a complex etiology. A large number of different processes are involved, such as cell migration, proliferation, differentiation, immune response and coagulation/fibrinolysis. Retinoids regulate a vast number of processes involved in it. CYP26B1 is one enzyme of P450-related retinoic acid hydroxylases that catabolise atRA to inactive metabolites and are important regulators of cellular retinoid levels. In this study we investigated rs2241057 SNP of CYP26B1 and the association with the risk of MI. Material and Methods: We genotyped 393 male patients with MI and 735 healthy male controls and 152 female patients and 273 healthy female controls by allelic discrimination using TagMan. Cross-sectional study of allelic frequencies was performed using Chi-Square Tests. Results: T-allel of rs2241057 SNP occurs more frequently in the cases of myocardial infarction compared to healthy controls in male population, C-allel is significantly less frequent in male patients compared to healthy controls (Pearson Chi-Square = 0,063, Fisher‘s Exact Test = 0,035). Multiple logistic analysis showed that C-allel was a protective factor for males with an odds ratio of 0,059. No difference was found between cases and controls among females. Conclusions: We found a strong tendency between frequency of occurrence of rs2241057 SNP C-allel and diminished number of cases of MI in male population, but not in the female population. Further research is needed to evaluate the role of CYP26B1 in cardiovascular disease. YI-804
NEW MARKERS OF OXIDATIVE STRESS AND ENDOTHELIAL DYSFUNCTION IN MYOCARDIAL INFARCTION
Y.I. Ragino, V.A. Baum, Y.V. Polonskaya, E.V. Sadovski. Institute of Internal Medicine SB RAMS, Novosibirsk, Russia Aim of the study was to investigate the relationships between oxidized
fibrinogen, oxidized apolipoproteins of low density lipoproteins (apo-LDL) and activity of plasma lipid peroxidation (LPO) process, endothelial dysfunction parameters and disturbances of hemostasis system in myocardial infarction (MI). 112 men 39-65 years old were included into the study: 43 group-1 men with MI in anamnesis last year, 19 group-2 men with subacute MI, 50 group-3 men without CHD. New methods for evaluation of oxidized fibrinogen and apo-LDL (patents of Russia) were worked out. Parameters of LPO process of plasma and LDL were determined as TBAreactive substances. Oxidative resistance of LDL was studied during LDL incubation with copper ions by spectrofluorimetrical method. Parameters of blood hemostasis system (antithrombine III, AT-III, thrombin inactivation index, TII, activity of XIIa-dependent fibrinolysis and other) and plasma level of NO metabolites were determined. Plasma levels of vonWillebrand factor and of endothelin-1 were evaluated by ELISA method. The extent of oxidized fibrinogen was 4 times higher (p<0,001) in group-2 men versus group-3 men. The level of oxidized apo-LDL was higher (p<0,01) in group-1 and group-2 men versus group-3 men. Independent association of oxidative modified fibrinogen with MI was revealed (p<0,001). Group-2 men had increased AT-III, TII and decreased activity of XIIa-dependent fibrinolysis as compared with group-3 men. Strong positive correlations between parameters of plasma LPO process and endothelial dysfunction and oxidized fibrinogen level were found (p<0,01). It is possible that oxidized fibrinogen may serve as biomarkers for MI. YI-805
SYNERGISTIC EFFECTS OF CHOLESTERYL ESTER TRANSFER PROTEIN, APOLIPOPROTEIN E, AND APOLIPOPROTEIN C3 GENE POLYMORPHISMS ON CORONARY ARTERY DISEASE RISK
A. Muendlein 1 , C.H. Saely 1,2 , S. Aczel 1,2 , T. Marte 1,2 , F. Schmid 1 , L. Koch 1 , P. Rein 1,2 , P. Langer 1 , H. Drexel 1,2,3 . 1 VIVIT Institute, Feldkich, Austria; 2 Academic Teaching Hospital Feldkirch, Feldkich, Austria; 3 University for Human Sciences, Triesen, Liechtenstein Background and aims: We aimed at investigating potential synergistic effects of cholesteryl ester transfer protein (CETP), apolipoprotein E (apoE), and apolipoprotein C3 (apoC3) gene variants on the risk of coronary artery disease (CAD). Methods: We analysed the CETP TaqIB, the APOE E3/E4/E2 and the APOC3 –482 C>T polymorphisms in 557 patients undergoing coronary angiography. Significant CAD was defined as the presence of coronary stenoses ≥50%. Results: From our patients, 129 carried the APOE E4 allele, 278 the APOC3 -482T allele and 196 the CETP B1B1 genotype. After multivariate adjustment, presence of the APOE E4 allele proved significantly associated with the presence of significant CAD (odds ratio 1.589 [95% CI 1.013-2.491]; p = 0.044); odds ratios for the APOC3 -482T allele and the CETP B1B1 genotype were 1.170 [0.813-1.686]; p = 0.398 and 1.453 [0.985-2.142]; p = 0.059, respectively. The risk of significant CAD was strongly increased in patients with more than one of the analysed variants: adjusted odds ratios were 1.831 [1.113-3.013]; p = 0.017 for patients with both the APOE E4 allele and the CETP B1B1 genotype, 1.983 [1.017-3.868]; p = 0.045 for patients with both the APOE E4 and the APOC3 -482T alleles, 2.498 [1.474-4.231]; p = 0.001 for patients with both the CETP B1B1 genotype and the APOC3 -482T allele, and 4.491 [1.222-16.499]; p = 0.028 for patients with all three variants. Conclusions: There are strong synergistic effects of the CETP TaqIB, APOE E3/E4/E2 and APOC3 –482 C>T polymorphisms on the risk of angiographically diagnosed CAD. YI-806
PCSK9 GENE ANALYSIS IN PORTUGUESE PATIENTS WITH FAMILIAL HYPERCHOLESTEROLEMIA
A.C. Alves 1 , Q. Rato 2 , A. Furtado 3 , M. Bourbon For The Portuguese Fam 1 . 1 Unidade de Investigacao Cardiovascular, Instituto Nacional de Saude Dr. Ricardo Jorge, Lisboa, Portugal; 2 Serv. de Cardiologia, Centro Hospitalar de Setubal, E.P.E., Hospital de S. Bernardo, Setubal, Portugal; 3 Serv. de Medicina Interna, Hosp. Pedro Hispano, Matosinhos, Portugal Background: Familial hypercholesterolemia (FH) is an autosomal dominant disorder associated with high risk of coronary heart disease. FH usually results from inherited defects in the Low Density Lipoprotein receptor gene (LDLR) and is characterised by increased circulating LDL cholesterol that leads to lipid accumulation in arteries and tendons (xanthomas), causing premature arteriosclerosis and coronary heart disease. Mutations
76th Congress of the European Atherosclerosis Society, June 10–13, 2007, Helsinki, Finland