Promoter polymorphism of macrophage Migration Inhibitory Factor MIF

Promoter polymorphism of macrophage Migration Inhibitory Factor (MIF) gene in Czech and Russian patients with myocardial infarction Petrkova J 1, 2, Tereshchenko IP 3, Lukl J 2, Mrazek F 2, Voevoda MI 3, Petrek M 2 1 Laboratory of Immunogenomics, Palacky University; 2 Dept. of Internal Medicine I, Faculty Hospital Olomouc, Czech republic 3 Inst. of Internal Medicine, RAMS Novosibirsk, Russian Federation This work was supported by Czech government (ME-856). I. T. is recipient of Visegrad fund fellowship)

Atherosclerosis & Inflammation Cytokine genetic variability • Inflammation of coronary artery wall is a critical process in the pathogenesis of myocardial infarction (MI). • Cytokines, namely proinflammatory, are implicated in atherosclerosis and pathogenesis of MI • Genes encoding for cytokines are polymorphic: some functional variants affect cytokine production • Cytokine polymorphism may be associated with MI susceptibility and/or manifestation

Macrophage Migration Inhibitory Factor • Cytokine with pleiotropic functions • Key regulator of inflammatory immune response • Expressed in macrophages, cardiomyocytes, vascular smooth muscle cells etc. • Regulates production of proinflammatory cytokines

MIF: candiate molecule in atherosclerosis • MIF was detected in early and advanced atherosclerotic lesions • MIF levels are increased in circulation of patients with acute myocardial infarction • Animal studies support role for MIF in pathogenesis of vascular disease • MIF gene (chr. 22 q 11. 2) is polymorphic, with a functional SNP in promotor and further candidate SNPs (e. g. in 3´flanking region)

AIMS • To investigate whether polymorphism in MIF gene promotor (-173 G/C) is associated with myocardial infarction in two Caucasian populations (Czech, Russian) • To explore if this polymorphism modifies manifestation of myocardial infarction • To study a role of another MIF single nucleotide polymorphism (rs 1007888) in MI susceptibility

Study population Diagnosis of myocardial infarction was made according to standard international criteria. (Eur Heart J. 2000; 21: 1502)

MIF polymorphism genotyping and statistical analysis • MIF -173 G/C (rs 755622) and MIF rs 1007888 polymorphism were genotyped by polymerase chain reaction with sequence specific primers (PCR-SSP) • Statistic: Statistical calculations were performed using the SPSS v. 13. 0 (SPSS Inc, Chicago, IL) The distribution of genotypes and alleles were analyzed using the Pearson’s 2 x 2 contingency table 2 -test and odds ratios and 95% confidence interval (CI) were estimated. The control populations was tested for conformity to the Hardy-Weinberg equilibrium.

RESULTS • There were no significant differences in MIF 173 G/C genotype, allelic and phenotype (carriage) frequencies between MI patients and controls for both populations (Table) • Regarding MIF-173 SNP, no differences after subgroup analysis (male / female) and age at 1 st MI episode) was found. • The proportion of MIF rs 1007888 GG homozygotes was higher among Czech female patients compared with female control group (Figure)

Table: Genotype, allele and phenotype (carriage rates) frequencies of the MIF 173 G/C polymorphism in the groups of patients with myocardial infarction and control subjects (in two investigated population) MIF -173 G/C Genotype % C allele carr. % Czech population (N=356) Russian population (N=414) MI (N=219) MI (N=240) Controls (N=137) Controls (N=174) GG 74. 4 75. 2 68. 3 72. 4 GC 21. 5 22. 6 30. 4 24. 1 CC 4. 1 2. 2 1. 3 3. 4 G 85. 2 86. 5 83. 5 84. 5 C 14. 8 13. 5 16. 5 15. 5 C 25. 6 24. 8 31. 7 27. 6 p (genotypes): CZ, 0. 61, RU, 0. 14; p (alleles): CZ, 0. 60, RU, 0. 72; p (carriage): CZ, 0. 87, RU, 0. 37.

Figure 1: Proportions of MIF rs 1007888 GG homozygotes in Czech patients with myocardial infarction and Czech control subjects in females (upper left) and males (bottom right) p females = 0. 027 ; p males = 0. 157

CONCLUSION • The data from two-population study do not support association between MIF-173 G/C polymorphism and susceptibility to / manifestation of myocardial infarction at least in Slavonic Caucasians. • A relationship between MIF SNP rs 1007888 and MI described for Czech females should be further explored.