Transcript Two RANTES gene polymorphisms and their - dr

Two RANTES gene polymorphisms
and their haplotypes in patients
with myocardial infarction from two
Slavonic populations
Petrek M1,Tereshchenko I1,3, Petrkova J2, Lukl J2, Mrazek F1,
Voevoda MI3
1Laboratory
of Immunogenomics, Palacky University; 2Dept. of
Internal Medicine I, Faculty Hospital Olomouc, Czech republic
3Inst. 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 & RANTES
• Inflammation of coronary artery wall is a critical
process in the pathogenesis of myocardial
infarction (MI).
• The chemokine RANTES/CCL5 activates and
attracts T-lymphocytes to inflammatory sites.
• RANTES detected in T-cells from atherosclerotic
lesions
• RANTES gene contains functional polymorphisms
thus representing candidate for MI susceptibility.
RANTES variants & Myocardial
infarction
• A single nucleotide polymorphism (SNP) locates at the position
-403 of the RANTES gene promoter (Guanin to Adenin -403
G→A)
RANTES-403*A allele tended to associate with a higher risk of coronary
artery disease (CAD) in Hungarian patients (Szalai et al. Atherosclerosis
2001;158:233-39).
In German population RANTES 403*A allele was significantly associated
with CAD (Simeoni et al. European Heart Journal 2004; 25,1438-46)
• Another SNP is located in intron of the RANTES gene (In1.1
T/C). This SNP was not studied in coronary atherosclerosis.
AIMS
• To investigate whether RANTES promoter -403
G/A and intronic In1.1 T/C polymorphisms are
associated with myocardial infarction in two
Caucasian populations (Czech, West-Slavonic;
Russian, East-Slavonic).
• In this way, to replicate previous reports on -403
SNP association with coronary atherosclerosis
from other populations and to extend the study
to another SNP in the RANTES gene.
Study population
Slavonic
population
West-Slavonic
East-Slavonic
Czech
Russian
140
224
patients with MI
unrelated
healthy
controls
197
244
patients with MI
unrelated
healthy
controls
Diagnosis of myocardial infarction was made
according to standard international criteria.
(Eur Heart J. 2000;21:1502)
RANTES polymorphism genotyping
and statistical analysis
• RANTES -403 G/A (rs2107538) and In1.1 T/C
(rs2280789) polymorphisms were genotyped by
polymerase chain reaction with sequence specific
primers (PCR-SSP)
• Statistics: Statistical calculations were performed using
the SPSS v.13.0 (SPSS Inc, Chicago, IL). The
distribution of the RANTES genotypes and alleles were
analyzed using the Pearson’s 2x2 contingency table 2test and odds ratios and 95% confidence interval (CI)
were estimated. The control populations was tested for
conformity to the Hardy-Weinberg equilibrium. Multiple
variable logistic regression model with adjustment for
conventional risk factors was used. Haplotypes were
estimated by maximum likehood (software ARLEQUIN, v.3.000).
RESULTS
• Both SNPs from all groups in Hardy-Weinberg equilibrium
• There were no significant differences in RANTES -403 G/A
genotype, allelic and phenotype (carriage) frequencies
between MI patients and controls for both populations (Tab.1)
• Also, no differences were observed for RANTES In1.1 T/C
SNP(Tab.2)
• Multivariate analysis with combined data from both populations showed no evidence of association, also after stratification by gender and age at 1st infarction episode (data not shown)
• There were no differences between RANTES haplotypes in
patients and control subjects (data not shown)
• The proportion of RANTES-403*A allele was similar in
control groups across several Caucasian populations (Figure)
Table 1: Genotype, allele and phenotype (carriage rates) frequencies of the
RANTES -403 G/A polymorphism in the groups of patients with myocardial
infarction and control subjects (in two investigated populations)
RANTES
-403 G/A
Genotype
%
Alleles
%
A allele
carriage %
a
Czech population (N=364)
Russian population(N=441)
MI
(N=224)
MI
(N=244)
Control
(N=140)
Control
(N=197)
GG
69.5
65.7
63.5
67.0
GA
27.8
31.4
29.9
28.9
AA
2.7
2.9a
6.6
4.1a
G
83.0
81.4
78.5
81.5
A
16.5
18.5b
21.5
18.5b
A
30.5
34.3c
36.5
33.0c
p value for genotype: Czech – 0.75, Russian – 0.48; b p value for alleles: Czech – 0.49, Russian –
0.27; c p value for A allele carriage: Czech – 0.45, Russian – 0.46.
Table 2: Genotype, allele and phenotype (carriage rates) frequencies of the
RANTES In1.1 T/C polymorphism in the groups of patients with myocardial
infarction and control subjects (in two investigated populations)
RANTES
Intron1.1 T/C
Genotype
%
Alleles
%
A allele
carriage %
Czech population (N=364)
Russian population(N=441)
MI
(N=224)
MI
(N=244)
Control
(N=140)
Control
(N=197)
TT
82.0
78.6
70.8
72.6
TC
16.2
21.4
25.5
25.8
CC
1.8
0
3.7
1.6
T
90.2
89.3
84.6
85.5
C
9.8
10.7
15.4
14.5
C
17.9
21.4
27.0
27.4
p values for all comparisons > 0.05
Figure: Proportions of RANTES -403*A allele in patient
with coronary artery disease and control groups across
selected populations
25
20
21,5
20,8
20
17,3
18,5
16,5
16,4
18,5
15
10
5
0
Germany
Hungary
Czech Rep.
patients
RF
control
Czech republic, Olomouc; Russian Federation (RF), Novosibirsk: myocardial
infarction, data of present study.
Germany:coronary atherosclerosis, Simeoni et al.,Eur Heart J.,2004;25:1438.
Hungary:coronary atherosclerosis, Szalai et al., Atherosclerosis.2001;158:233.
CONCLUSIONS
• RANTES promoter and intronic polymorphisms, and their
haplotypes, are not associated with susceptibility to
myocardial infarction and do not act as disease modifiers.
• Further, our data do not support association between
RANTES -403*A allele and MI as previously reported from
LURIC (LUdwigshafen Risk and Cardiovascular health)
and Hungarian cohorts.
• At least in Slavonic population, RANTES promoter
- 403 G/A and intronic In1.1 T/C polymorphism do not
contribute to genetic determination of myocardial
infarction.