2008 Barcelona - European Society of Human Genetics

Molecular and biochemical basis of disease
P06.307
Genome-wide screen for aberrant DNA methylation in human
uterine leiomyoman by ms-RDA
S. Izumi 1 , L. Cai 1 , M. Abe 2 , A. Kondo 1 , Y. Morita 3 , M. Mizoguchi 3 , N. Uchida 1 , S.
Takagi 3 , T. Ushijima 2 ;
1 Department of Obstetrics and Gynecology, Tokai University School of Medicine,
Isehara, Japan, 2 Carcinogenesis Division, National Cancer Center Research
Institute, Tokyo, Japan, 3 Department of Clinical Genetics, Tokai University
School of Medicine, Isehara, Japan.
Uterine leiomyoma are very common benign tumors in women of reproductive
age . However the molecular mechanisms of cause and development
of these tumors are poor understood . This study is attempted
to examine whether aberrant DNA methylation occurred in these
tumors . Therefore we carried out a genome-wide screen for aberrant
DNA methylation, adopting methylation-sensitive-representational difference
analysis (MS-RDA) using normal adjacent myometria as tester
and myoma tissue driver . As the results, a total of 192 clones were
sequenced, 27 DNA fragments derived from CpG islands (CGIs) were
isolated, and seven of them were from CGI in the 5’ regions of known
genes, which include CHARC1, FAM44B, FLJ33655, HSUP, MLLT3,
SLC16A1, and ZNF96 . Then, methylation statuses of those CGIs
were analyzed by methylation-specific PCR (MSP) using five primary
samples of human uterine leiomyoma . Aberrant DNA methylation did
not observed in seven genes in five human uterine leiomyoma eventually
. Though, this study failed to identify aberrant DNA methylation occurring
in the human uterine leiomyoma by a genome-wide screen, it
dose not exclude that epigenetic modifications of DNA methylation are
involved in the cause and development of uterine leiomyomas . A large
population of primary samples and more attempts, such as use cell
lines or primary monolayer cultures established from tissue samples,
are needed be done to develop this issue
P06.308
study of the VEGF Polymorphism in HELLP syndrome patients
B. Nagy 1 , A. Molvarec 1 , T. Várkonyi 1 , B. Rigó 1 , H. Savli 2 , P. Hupuczi 1 , J. Rigó
Jr 1 ;
1 Semmelweis University, Budapest, Hungary, 2 Kocaeli University, Izmit, Turkey.
Background: The vascular endothelial growth factor (VEGF) has a
critical role in vasculogenesis and vascular permeability in several
diseases including preeclampsia . VEGF G+405C, C-2578A and C-
460T SNPs are known to be related to VEGF production . We decided
to determine the allele and genotype frequencies of VEGF G+405C,
C-460T and C-2578A SNPs in healthy pregnant women and HELLP
syndrome patients .
Methods: The authors introduced a quantitative real-time PCR method
for the determination of the three VEGF SNPs . Blood samples were
collected from 71 HELLP syndrome patients and 93 healthy controls .
Results: There were significant differences in the allele and genotype
frequencies of VEGF C-460T SNP between the two study groups . The
T allele was present in 71 .1% in the HELLP group, while in 53 .8% in
the controls (p=0 .0014) . The TT genotype occurred significantly more
frequently in the HELLP group than in the control group (45 .1% vs .
21 .5%; p (for genotype frequencies)=0 .0011) . The TT genotype carriers
had an increased risk of HELLP syndrome, which was independent
of maternal age and primiparity (adjusted odds ratio (OR)=3 .03,
95% confidence interval (CI)=1.51-6.08; p=0.002). Although the VEGF
G+405C allele and genotype distributions did not differ significantly
between the two groups, the CC genotype carriers were also found
to have an increased risk for HELLP syndrome after adjustment for
maternal age and primiparity (adjusted OR=3 .67, 95% CI=1 .05-12 .75;
p=0 .041) . The VEGF C-2578A SNP was not associated with HELLP
syndrome .
Conclusions: We found that the VEGF -460TT and +405CC genotype
carriers have an increased risk of HELLP syndrome .
P06.309
VEGF A2578C polymorphism is associated with muscle fiber
type distribution in athletes
E. V. Lyubaeva 1 , I. I. Ahmetov 2 , A. M. Hakimullina 2 , O. L. Vinogradova 1 , V. A.
Rogozkin 2 ;
1 SRC Institute for Biomedical Problems of the Russian Acad. Sci., Moscow,
Russian Federation, 2 St Petersburg Research Institute of Physical Culture, St
Petersburg, Russian Federation.
There is strong relationship between muscle fiber type distribution
and human physical performance . For example, elite weightlifters and
sprinters exhibit large percentages of fast-twitch fibers (FT, also known
as type II muscle fibers) compared to controls and endurance athletes.
FT fibers have comparatively low capillary density and blood flow capacity
and low mitochondria content . Vascular endothelial growth factor
(VEGF) is important in the basal maintenance of skeletal muscle
capillarization and may influence the determination of muscle fiber
type distribution. To investigate the question of the influence of VEGF
gene polymorphism on the proportion of fibers types of m. vastus lateralis,
we have analyzed the muscle biopsies obtained from 21 elite
Russian athletes (all-round speed skaters) . The immunoperoxidase
technique was employed for immunohistochemical identification of
myosin isoforms . VEGF gene A2578C polymorphism was determined
by PCR-RLFP. Mean percentages of FT fibers were significantly higher
in VEGF CC homozygotes than in VEGF A allele carriers (AA/AC -
38 .3 (6 .2) %, CC - 47 .8 (12 .4) %; P=0 .03) . Then we determined distribution
of VEGF alleles in 60 elite and sub-elite weightlifters and in
1,113 controls . We found that the frequency of VEGF 2578C allele was
significantly higher in weightlifters than in controls (58.3% vs. 48.0%;
P=0 .035) . In conclusion, VEGF 2578C allele is associated with increased
proportion of FT muscle fibers in all-round speed skaters and
with elite power athlete status .
P06.310
VEGF iD polymorphism and peripheral neuropathy susceptibility
M. Stavarachi1 , D. Cimponeriu1 , P. Apostol1 , M. Toma1 , I. Radu1 , A. Craciun2 , D.
Cheta2 , L. Gavrila1 ;
1 2 University of Bucharest, Institute of Genetics, Bucharest, Romania, ”N.
Paulescu” Institute, Bucharest, Romania.
Vascular endothelial growth factor (VEGF) is a cytokine involved in angiogenesis
and in maintaining of neuronal function . The genetic variation
in VEGF promoter may be responsible for different neuropathies,
including the diabetic one .
The aim of our study was to investigate the relationship between the
-2549 I/D VEGF gene promoter polymorphism and diabetic peripheral
neuropathy susceptibility .
A case-control study was performed in 60 diabetic patients with peripheral
neuropathy and 60 healthy subjects, sex and age matched .
After the informed consent was obtained, DNA was extracted from peripheral
blood . The VEGF -2549 I/D genotyping were assessed using
PCR method .
The distribution of VEGF -2549 ID in both lots respected the Hardy-
Weinberg condition . The frequencies of genotypes and alleles in the
patients and healthy control groups were compared using the contingency
tables . Although the DD genotype and D allele frequencies
were higher in diabetic patients than in controls (DD: 33 .3% vs 25%, D:
57 .5% vs 52 .5%) these differences do not reach the statistical power .
We also observed that the trend of association increases if only patients
with early onset were compared (OR~1,2) .
This preliminary study indicates that the VEGF ID polymorphism may
be associated with early onset diabetic peripheral neuropathy . A future
study with an increased statistical power is necessary to clarify the
relationship between this polymorphism and peripheral neuropathy
susceptibility .
The research was funded by GAR 183/ 2007 .
P06.311
Linkage and association study for VEGF serum level in the
isolated population of campora
D. Ruggiero1 , R. Sorice1 , C. Bellenguez2 , T. Nutile1 , G. Fardella1 , M. Aversano1 ,
C. Bourgain2 , M. Ciullo1 ;
1Institute of Genetics and Biophysics A. Buzzati-Traverso, CNR, Naples, Italy,
2INSERM U535, Villejuif, France.
Vascular Endothelial Growth Factor (VEGF) is an angiogenic factor
having a key role in both physiological and pathological angiogenesis .
In order to identify QTLs influencing VEGF serum levels, we performed
a genome-wide linkage analysis using a sample of individuals randomly
chosen in Campora, an isolated population in South Italy .
VEGF serum levels were measured using ELISA method in a sample of
656 individuals . 627 individuals out of the 656 were all related through
a 3049-member pedigree and genotyped for 1122 microsatellites on