Deep vein thrombosis (DVT) is a multifactorial disease caused by interactions between acquired risk factors and coagulation abnormalities.1  In whites, the factor V–Leiden and the prothrombin-20210G>A are widely recognized as genetic risk factors for DVT. However, these 2 mutations are not present in Japanese populations, and little is known about the genetic risk factors for DVT in these populations. In this study, we evaluated the genetic contributions of 5 polymorphisms in Japanese DVT patients. The plasminogen-A620T mutation, formerly referred to as plasminogen-Tochigi, and the protein S–K196E mutation, formerly referred to as protein S–Tokushima, exhibited decreased activities of plasminogen and protein S despite normal antigen levels.2-4  The ADAMTS13-P475S mutation exhibited low von Willebrand factor–cleaving activity in vitro.5  The factor XII–4C>T substitution in the 5′-untranslated region, formerly referred to as 46C>T, showed decreased plasma levels of both antigen and activity.6  The plasminogen activator inhibitor-1 (PAI-1) 4G/5G polymorphism is related to in vitro differences in transcription activity.7  We genotyped subjects for these 5 polymorphisms and compared their genotypic frequencies between 161 DVT patients and 3655 population-based controls. The protocol for this study was approved by the ethical review committee, and only those subjects who provided written informed consent for genetic analyses were included in this study. All participants of this study were Japanese. The controls were from a general population randomly selected from the residents of Suita City located in the second largest urban area in Japan (the Suita Study).8  One hundred sixty-one DVT patients, 78 men and 83 women, were registered by the Study Group of Research on Measures for Intractable Diseases, working under the auspices of the Ministry of Health, Labor, and Welfare of Japan. Six centers (Tochigi, Tokyo, Nagoya, Kyoto, and 2 in Osaka) participated in this study. The patients' mean age was 49.5 years (range, 12-87 years) and their mean body mass index was 23.6 ± 3.3. Thirteen percent of patients had a family history of thrombosis, and 16% of the patients had recurrent thrombosis.

Of all the polymorphisms tested, only the frequency of protein S–K196E was statistically different between the 2 groups (χ2 = 38.3, P < .001) (Table 1). No other frequency differences were statistically significant. Two DVT patients were homozygous for the protein S–196E allele; however, no homozygotes were identified in the control group. One patient with the 196EE genotype first developed DVT following surgery at age 47, while the other patient developed DVT during pregnancy at age 32.

Table 1.

Numbers and genotypic frequencies of protein S–K196E mutation in the DVT and control groups

Genotypes
General population, no. (%)
DVT group, no. (%)
Additive model*   
   KK   3585 (98.2)   146 (90.7)  
   KE   66 (1.8)   13 (8.1)  
   EE   0 (0.0)   2 (1.2)  
   Total   3651 (100.0)   161 (100.0)  
Dominant model†   
   KK   3585 (98.2)   146 (90.7)  
   KE + EE   66 (1.8)   15 (9.3)  
   Total
 		 3651 (100.0)
 		 161 (100.0)
 
Genotypes
General population, no. (%)
DVT group, no. (%)
Additive model*   
   KK   3585 (98.2)   146 (90.7)  
   KE   66 (1.8)   13 (8.1)  
   EE   0 (0.0)   2 (1.2)  
   Total   3651 (100.0)   161 (100.0)  
Dominant model†   
   KK   3585 (98.2)   146 (90.7)  
   KE + EE   66 (1.8)   15 (9.3)  
   Total
 		 3651 (100.0)
 		 161 (100.0)
 

DNA genotyping was performed by the TaqMan allele discrimination method.9  We have adopted the numbering standards of the Nomenclature Working Group, wherein the A of the ATG of the initiator Met codon is denoted as nucleotide + 1, and the initial Met residue is denoted as amino acid + 1, resulting in the renaming of several mutant alleles.10  Comparisons between the DVT cases and the controls were analyzed using a χ2 test with the genotypes as independent variables (indicated by P and OR) or using multiple logistic analyses with the genotypes as independent variables and age and sex as covariates (indicated by P′ and OR′).

*

For comparison of general population to DVT group, P was not determined

†

For comparison of general population to DVT group, P < .001; OR = 5.58 (3.11-10.01); P′ < .001; OR′ = 4.72 (2.39-9.31)

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The mutant protein S with the E allele has already been intensively studied as protein S–Tokushima.11  The protein S mutant showed the reduced activated protein C cofactor activity compared with wild-type protein S, suggesting a direct link between the protein S–K196E mutation and the development of DVT. By the genotyping of the general population, the protein S–196E allele frequency was estimated as 0.009. Thus, a substantial portion of the Japanese population harbors this mutant allele and is at higher risk for DVT.

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2006
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