1、Toshiyuki MIYATA, PhD National Cardiovascular Center Research Institute, Osaka, Japan the 19th Great Wall International Congress of Cardiology, October 24, 2008 Genetic risk factors for venous thromboembolism in Japanese Advanced age is a risk for venous thromboembolism (VTE) All VTE Blood, 110, 309
2、7-3101, 2007 among residents of Olmsted County, Minnesota, from 1966 to 1990 PE DVT alone Male Female Incidence rates of VTE increase dramatically at about age 55, and, by age 80, are nearly 1 in 100 per year, approximately 1000-fold higher than for those aged 45 or younger. Risk factors for venous
3、thromboembolism N Engl J Med, 358, 1037-52, 2008 Acquired factors Reduced mobility Advanced age Cancer Acute medical illness Major surgery Trauma Spinal cord injury Pregnancy and postpartum period Polycythemia vera Antiphospholipid antibody syndrome Oral contraceptives Hormone-replacement therapy He
4、parins Chemotherapy Obesity Central venous catheterization Immobilizer or cast Hereditary factors Antithrombin deficiency Protein C deficiency Protein S deficiency Factor V Leiden Prothrombin gene mutation Thrombo- modulin endothelium EPCR PC PS prothrombin FVa FXa FVa FVIIIa HSPG thrombin APC Prote
5、in C/protein S system Proteoglycan sulfate system Heparan sulfate FVi FVIIIi IIa AT Anticoagulant Systems on Endothelium Caucasian Japanese Factor V Leiden Prothrombin G20210A Deficiencies of protein C, protein S, antithrombin Low frequency variation Deficiencies of protein C, protein S, antithrombi
6、n Very rare mutation Genetic risk factors for venous thromboembolism Protein S K196E Tokyo General population, Suita study at NCVC VTE patients: Sub-group of Blood Coagulation Abnormality Study Group Suita, Osaka NCVC, Osaka U Nagoya U Jichi Med U Keio Univ Kyoto Pref Med U About 170 Japanese VTE pa
7、tients Case-control study for VTE more than 3,500 individuals from a general population randomly selected from Suita city residents Kimura et al., Blood, 2006 VTE 13920 1 160 0.069 3290 332 17 3639 0.050 2.179 0.336 Pro Ser Major homo Hetero Minor homo Total MAF Major homo Hetero Minor homo Total MA
8、F 2 P Major allele Minor allele General population (Suita study) 152 9 0 161 0.028 3501 149 0 3650 0.020 0.987 0.320 Ala Thr 146 13 2 161 0.053 3585 66 0 3651 0.009 75.464 T 61 69 30 160 0.403 1468 1686 497 3651 0.367 3.402 0.183 4G 5G ADAMTS13 P475S PLG A620T PS K196E 63 75 23 161 0.376 1513 1651 4
9、86 3650 0.359 0.372 0.830 T C PAI-1 4G/5G MAF: minor allele frequency Case-control association study using Japanese VTE patients Protein S K196E as a Risk for VTE Population based-control n (%) Geno- types VTE group n (%) Protein S K196E (Protein S Tokushima) KK KE EE Total KK KE+EE total 3585 (98.2
10、) 66 (1.8) 0 (0.0) 3651 (100.0) 3585 (98.2) 66 (1.8) 3651 (100.0) 2 = 75.464, P0.0001 2 = 41.807, P0.0001 OR=5.58 (95% CI 2.90-9.46) 146 (90.7) 13 (8.1) 2 (1.2) 161 (100.0) 146 (90.7) 15 (9.3) 161 (100.0) OR: odds ratio, CI: confidence interval Gla EGF-like 1-4 Sex hormone-binding globulin 635 aa Pr
11、otein S K196E Protein S Tokushima Lys196Glu PS K196E mutation has been identified in Japanese DVT patients in 1993 by two independent groups. This mutation was referred to as protein S Tokushima. It is present in the second EGF-like domain of protein S. In vitro study showed that this mutant exhibit
12、s the loss of the APC cofactor activity and the prothrombinase inhibitory activity. Yamazaki et al, 1993, Shigekiyo et al, 1993, Hayashi et al, 1994 Tokyo Nagoya U 3 hetero / 182 individuals Heterozygosity: 1.65% Allele freq.: 0.82% Kyushu U 5 hetero / 304 individuals Heterozygosity: 1.64% Allele fr
13、eq.: 0.82% Geographical distribution of PROS1 K196E mutation Natl Cardio Vasc Ctr at Suita 66 heterozygotes / 3,651 individuals Heterozygosity: 1.81% Allele freq.: 0.90% Heterozygotes N=34 Wild-types N=1,828Staclot Protein S(Diagnostica Stago) Protein S activity in wild-type and K196E heterozygous i
14、ndividuals 71.9+/-17.6% 87.9+/-19.8% Protein S activity P0.0001 Kimura et al., JTH, 4, 2010-2013, 2006 Acquired factors influencing plasma protein S activity Gender and age dependency of plasma protein S levels Sakata et al, JTH 2004 Men N=1252 Women N=1438 30 40 50 60 70 80 140 120 100 80 60 Protei
15、n S activity (%) Age band Age Gender Pregnancy Oral contraceptives Anticoagulant drugs DIC Liver diseases Kidney disease 1. PROS1 K196E mutation is a genetic risk factor for venous thromboembolism (VTE) in Japanese. 2. A missense mutation causing Lys196 to be replaced by Glu is located within the 2n
16、d EGF-like domain. 3. The odds ratio of the mutant E allele for VTE was 5.58. 4. Individuals heterozygous for the mutant E-allele had lower (mean 16%) plasma protein S activity than wildtype subjects. PROS1 K196E Kimura et al., Blood, 2006, Kimura et al., JTH 2006, Miyata et al., IJH 2006 5. The all
17、elic frequency of the E allele was 0.9%. We estimated a total of as many as 10,000 Japanese as homozygotes. 6. A substantial proportion of the Japanese population carries the PROS1 E allele and is at risk of developing VTE. 7. Therefore, individuals with the PROS1 E allele should avoid environmental
18、 risk factors known to be associated with VTE. PROS1 K196E, cont. Kimura et al., Blood, 2006, Kimura et al., JTH 2006, Miyata et al., IJH 2006 Caucasian Japanese Factor V Leiden Prothrombin G20210A Deficiencies of protein C, protein S, antithrombin Low frequency variation Deficiencies of protein C,
19、protein S, antithrombin Very rare mutation Genetic risk factors for venous thromboembolism Protein S K196E Methods: We have sequenced the entire coding regions of 3 genes in all 173 DNA samples obtained from Japanese VTE patients without any consideration of their activities and antigen levels. ABI
20、3730 DNA Analyzer Question Prevalence of nonsynonymous mutations in PROS1, PROC, and SERPINC1 (antithrombin) in the Japanese VTE patients Number of Mutation Carriers 55 out of 173 VTE patients (32%) carried nonsynonymous mutations SERPINC1 14 5 PROS1 24 PROC 12 Including one patient with PROS1 gene
21、deletion nonsynonymous mutations Missense, Nonsense, Frame-shift, Splice-site, Inframe deletion PS K196E mutation as a modifier R221W V339M V339M R271W K193del K196E K196E K196E K196E K196E No Yes No Yes unavailable 40 25 55 39 57 PROCPROS1 Family history Onset age of initial VTE 5UTRpromoter centro
22、mereD3S3619D3S3634 Chromosome 3 Large PROS1 deletion in 1 out of 163 VTE patients 3 4 7 11 1321 56 89 12 14 10 15 PROS1 Deletion, at least 107 kb Yin et al., Thromb Haemost 2007; 98: 783-789 A large PROS1 deletion was found in one VTE patient who showed 16% PS activity and did not have point mutatio
23、ns in PROS1. Comparison of first onset age of VTE events between mutation carriers and non-carriers 55* 44.7+/-16.5 118 52.6+/-16.1 Number P=0.0031 Carriers Non-carriers First onset age mean+/-SD * Five had mutations in both PROS1 and PROC. Two were homozygotes for PROS1 K196E. One was a compound he
24、terozygote for PROS1 K196E/R101C. Recurrent mutations found in Japanese VTE patients PROS1 K196E in the 2nd EGF-like domain 2 homo, 13 hetero, reduced PS activity with normal PS antigen PROC K193del at the 6th residue from the C- terminus in the light chain 4 hetero, normal PC amidolytic activity wi
25、th reduced anticoagulant activity PROC V339M in the catalytic 4 hetero, reduced PC amidolytic activity only found in Japanese population only found in Japanese population SERPINC1 14 Other genetic factors 5 PROS1 24 PROC 12 VTE Genetic risk factors Environmental risk factors Obesity, Cancer, Immobil
26、ization, Hospitalization, Surgery, Pregnancy CLOT prevention Risk factors for venous thromboembolism N Engl J Med, 358, 1037-52, 2008 Acquired factors Reduced mobility Advanced age Cancer Acute medical illness Major surgery Trauma Spinal cord injury Pregnancy and postpartum period Polycythemia vera
27、Antiphospholipid antibody syndrome Oral contraceptives Hormone-replacement therapy Heparins Chemotherapy Obesity Central venous catheterization Immobilizer or cast Hereditary factors Antithrombin deficiency Protein C deficiency Protein S deficiency Factor V Leiden Prothrombin gene mutation Acknowled
28、gements Natl Cardiovascular Ctr Res Inst Drs. Tong YIN, Rina KIMURA, Koichi KOKAME, Shigenori HONDA, Yongchol SHIN, Ms. Junko ISHIKAWA, Yukiko SATO The Study Group of Blood Coagulation Abnormality on Measures for Intractable Diseases Drs. Tomio KAWASAKI, Hajime TSUJI, Seiji MADOIWA, Yoichi SAKATA, Tetsuhito KOJIMA, Mitsuru MURATA, Yasuo IKEDA Natl Cardiovascular Ctr Hospital Drs. Yoshihiro KOKUBO, Toshiyuki SAKATA, Satoshi TAKESHITA, Hitonobu TOMOIKE
