P111 LEPTIN TO ADIPONECTIN RATIO IS MODIFIED BY ADIPONECTIN RECEPTOR GENES POLYMORPHISM AND DIET

78th EAS Congress

Atherosclerosis Supplements 11, no. 2 (2010) 17–108

CAD/MI (rs9818870 P = 7.44×10−13 (OR = 1.15; 95% CI: 1.11–1.19) by GWAS. The underlying biology is unclear. We aimed to perform a MRAS gene based pathway analysis using Affymetrix genotype data from the German MI Family Studies [GerMIFS I, GerMIFS II, and GERMIFS III (KORA)] comprising a total of 2,000 MI cases and 3,000 population-based controls. Methods: Identification of relevant genes was undertaken by bioinformatical means of medical literature and internet databases. Available SNPs were investigated in silico in three independent genome-wide SNP data sets genotyped on Affymetrix platforms; missing genotypes were subsequently imputed (GerMIFS I [875 MI cases, 1644 controls] and GerMIFS II [1221 MI cases, 1298 controls]. Results: Thirteen genes were identified in TNFa-M-Ras-LFA-1 pathway. Among them, ten genes carried several SNPs significantly associated with CAD/MI in both GWAS. In ICAM-1 17/70 SNPs, in ITGAL 8/47 SNPs, in ITGB2 25/147 SNPs, in MRAS 43/91 SNPs, in Rap1A 29/276 SNPs, in RAPL 30/194 SNPs, in RIAM 16/228 SNPs, TNFa 13/140 SNPs, in TNFRSF1A 7/56 SNPs and in TNFRSF1B 14/91 SNPs were significant for CAD/MI [combined p-values ranging from 0.001 to 5×10−5 ). Conclusions: Several SNPs in genes of the TNFa–M–Ras–LFA-1 pathway revealed significant results for CAD/MI in two GWAS. Replication steps are needed to further establish the role of this pathway in the etiology of CAD/MI. P108 MOLECULAR GENETIC ANALYSIS OF A HUMAN IGF1 PROMOTER P1 VARIATION 2 R. Telgmann1 , C. Dordelmann ¨ , E. Brand3 , V. Nicaud4 , H. Pavenstadt ¨ 3, F. Cambien4 , L. Tiret4 , M. Paul5 , S.-M. Brand-Herrmann1 . 1 Molecular Genetics of Cardiovascular Disease, Leibniz-Institute for Arteriosclerosis Research, 2 Institute for Analytical Sciences, Dortmund, 3 University Clinic Munster, ¨ Munster, ¨ Munster, ¨ Germany, 4 INSERM UMRS 937, Paris, France, 5 Maastricht University, Maastricht, The Netherlands IGF1 exerts important endocrine and paracrine functions in the cardiovascular system. We identified the common variant C-1411T in the IGF1 upstream promoter P1, located within several overlapping transcription factor binding sites (TFBS). Using transient transfection assays, we identified this site as a functional enhancer of several, partly overlapping conensus binding sites. The T allele-carrying enhancer, compared to the C allelic portion, exerts significantly reduced or even abrogated activity in SaOs-2 and HepG2, respectively (Pvalues < 0.0001), as well as in differentiated THP-1 macrophages. EMSA and subsequent Supershift experiments in HepG2 identified c-Jun as binding partner exclusively to the T allele, whereas C/EBPdelta and ICSBP/IRF8 interacted only with the C allelic promoter portion. Furthermore, genotyping of a case–control study for essential hypertension (n = 745 hypertensives; n = 769 normotensives) for this variant revealed an odds ratio for hypertension of 0.73 ([95% CI 0.58– 0.91], P = 0.006) associated with the T allele, and normotensives carrying the protective T allele displayed a significant decrease in diastolic (P = 0.036) and systolic (P = 0.024) blood pressure level. We here report detection of a functional enhancer module in the upstream IGF1 promoter region, which might play a key role in local IGF1 bioavailability. Whether C-1411T is also associated with other IGF1-related disease phenotypes should be evaluated further population studies. P109 FUNCTIONAL ANALYSES OF THE HUMAN LEUKOTRIENE C4 SYNTHASE GENE PROMOTER MOLECULAR HAPLOTYPES F. Bruns1 , R.-G. Telgmann1 , B. Schroer ¨ 1 , B. Schmitz2 , F. Cambien3 , ¨ P. Amarenco4 , E. Brand2 , S.-M. Brand-Herrmann1 . 1 Leibniz-Institut fur ¨ Arterioskleroseforschung, 2 Universitatsklinikum Munster, ¨ Munster, ¨ Germany, 3 INSERM UMRS 937, 4 INSERM U476UMRS 698, Paris, France LTC4 Synthase (LTC4 S) SNPs have been validated to be associated with cardiovascular disease (CVD), especially stroke phenotypes. LTC4 S is a strong mediator of inflammation and affect vascular permeability, its expression being strictly limited to hematopoietic cells. We aimed at analysing the molecular basis of LTC4 S promoter control and identifying functional genetic variants with molecularly proven consequences for LTC4 S expression. LTC4 S was genetically scanned in 120 chromosomes of high-risk CVD patients (MolProMD Study). Molecular haplotypes (MolHaps) were identified by individual DNA subcloning. Promoter deletion constructs were generated and transfected in THP1 and U937 cells, the different allelic constellations were introduced by site-directed mutagenesis. We identified four SNPs (−1783G>A, −1412C>T, −1008G>A, −380A>C) in the 5’-regulatory region, generating six common MolHaps. We found that 2010 bp of the LTC4 s 5 flanking region were transcriptionally active in monocytic cell lines, suggesting a cis-active regulatory element located between positions −2010 and −1782 from the TIS. In U937 cells, a second region between positions −1782 and −1560 displayed high transcriptional activity. Transfection analyses of promoter fragments harbouring different MolHaps indicated that MolHaps2 (P < 0.01; harbouring the functional SNP −380A>C) and 5 (P < 0.01; harbouring

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the cis-located −1783G>A) were significantly less transcriptionally active in THP1 and U937 cells compared to wild type MolHap1. We identified ~2010 bp of the LTC4 S 5 flanking region to be sufficient for meaningful transcriptional activity in monocytic cell lines, mapped cis-active regulatory elements and provide a basis for the molecular functional role of −380A>C in CVD phenotype association. P110 CORONARY ARTERY CALCIUM SCORE IS A MORE SENSITIVE INDICATOR OF SUBCLINICAL ATHEROSCLEROSIS THAN ANKLE-BRACHIAL INDEX A. Kalvelis1 , I. Stukena1 , G. Bahs1 , L. Zvaigzne2 , A. Lejnieks1 . 1 Riga Stradins University, 2 Medical Company ‘ARS’, Riga, Latvia Aim: To assess the amount of calcium in coronary arteries (CACscore) by using CT scan and ankle-brachial index (ABI), and their correlations with risk factors of cardiovascular disease (CVD) in patients with adiposity. Patients and Methods: In 200 patients [mean age 52.3±11.7 (±SD)] CACscore by CT scan and ABI were measured. The correlations of these parameters with waist circumference (WC), body mass index (BMI), systolic and diastolic blood pressure (SBP and DBP), total cholesterol (TC), low-density and high-density lipoprotein cholesterol (LDL-C and HDL-C), triglycerides (TG), glucose, C-reactive protein (CRP), adiponectin, and leptin were analyzed in accordance with the Pearson correlation coefficient. Results: In the whole group mean ABI was 0.99±0.14 and CACscore 120±403 without substantial difference between men and women. In persons with adiposity, elevated CACscore was detected, while ABI did not show difference between persons with or without signs of adiposity. ABI and CAC score in different patient subgroups are shown in table. Only in patients with adiposity CACscore had significant correlation with glucose, TC, LDL-C, HDL-C, and adiponectin. Only in patients with increased WC, ABI correlated with SBP and TC while in patients with elevated BMI, ABI did not correlated with any of analyzed parameters. Conclusions: Elevated CACscore is a more sensitive indicator of CVD risk which correlates with changes of levels of glucose, lipids, and adiponectin. This association is more marked in patients with adiposity. Table: ABI and CAC score in different patient subgroups BMI Normal

WC Elevated

p

Normal

Whole group Increased

p

ABI

1.01 (0.93, 1.07)

1.00 (0.94, 1.06)

0.457

1.02 (0.94, 1.09)

1.00 (0.94, 1.06)

0.210

1.00 (0.93, 1.06)

CAC score

0 (0, 31)

7 (0, 100)

0.007

0 (0, 1)

2 (0, 61)

0.013

0 (0, 33)

P111 LEPTIN TO ADIPONECTIN RATIO IS MODIFIED BY ADIPONECTIN RECEPTOR GENES POLYMORPHISM AND DIET I. Wybranska1 , M. Malczewska-Malec1 , I. Leszczynska-Golabek1 , B. Kiec-Wilk1 , M. Kwasniak1 , C. Defort2 , J. McMonagle3 , H. Roche3 , A. Dembinska-Kiec1 . 1 Department of Clinical Biochemistry, Jagiellonian University Collegium Medicum, Cracow, Poland, 2 INSERM U476, Marseille, France, 3 University College Dublin, Dublin, Ireland Background: The leptin to adiponectin ratio has been suggested to be predictor of development of metabolic syndrome (MetS) complications. The present study was undertaken to investigate whether the dietary intervention can predict the risk of traits characterizing MetS and affect the impact of leptin/adiponectin ratio in the adiponectin receptor 1 (ADR1) gene variants (rs2275737; rs 10920533) and adiponectin receptor 2 gene variant (rs 1058322) (ADR2) carriers. Material and Methods: The present study was the part of EU LIPGENE human dietary intervention project in patients with MetS. The 486 MetS subjects were randomly assigned to one of four isoenergetic diets: high-fat (38% energy) high-SFA (Diet A); high-fat (38% energy), highMUFA (Diet B) and two isoenergetic low-fat (28% energy) diets, supplemented with long chain n-3 PUFA (Diet D) or it placebo (Diet C) for 12 weeks. Results: However before dietary intervention there was any statistical differences in the leptin/adiponectin ratio value between studied genotypic subgroups, the moderate (about 10%) decrease of leptin/adiponectin ratio after 12 weeks of low-fat diet (D and C) was observed in common genotype carriers of ADR1 and ADR2. The significant increase in leptin/adiponectin ratio in the rare AA genotype (rs 109205330) 17.75 vs 9.34 (GG) and rare AA genotype (rs 2275737) 16.01 vs 9.73 (CC) was observed. Conclusion: Low-fat, high carbohydrate diet (8% SFA, 11% MUFA; 6% PUFA), enriched in n-3 PUFA is beneficial for MetS, but not with the AA genotype carriers of adiponectin rs109205330 and rs2275737. Supported by the EU FOOD-CT-2003–505944 LIPGENE and K/ZDS/000620 projects