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浙江大学学报(医学版)  2018, Vol. 47 Issue (6): 606-611    DOI: 10.3785/j.issn.1008-9292.2018.12.07
专题报道     
遗传性易栓症筛查及相关基因检测分析
尹黎1(),李歌2,沈健3,刘震杰1,*()
1. 浙江大学医学院附属第二医院血管外科, 浙江 杭州 310009
2. 浙江大学医学院附属邵逸夫医院病理科, 浙江 杭州 310006
3. 浙江大学医学院附属第二医院心血管研究所, 浙江 杭州 310009
Screening for inherited thrombophilia and genome sequencing
YIN Li1(),LI Ge2,SHEN Jian3,LIU Zhenjie1,*()
1. Department of Vascular Surgery, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China
2. Department of Pathology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China
3. Institute of Cardiovascular Diseases, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China
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摘要:

目的: 检测和分析遗传性易栓症的相关基因及其突变位点,为中国人群遗传性易栓症患者的基因突变特点积累资料。方法: 对2015年6月至2017年12月在浙江大学医学院附属第二医院就诊的遗传性易栓症疑似患者25例进行血清蛋白C、蛋白S,抗凝因子相关基因蛋白C(基因名PROC)、蛋白S(基因名PROS1)和抗凝血酶Ⅲ(基因名SERPINC1),凝血因子相关基因凝血因子Ⅴ(基因名F5)、凝血因子Ⅱ(基因名F2)、凝血因子Ⅷ(基因名F8)、高型半胱氨酸血症相关因子(CBSMTHFR)第二代测序分析。利用千人基因组数据库、ESP6500数据库、Genoma数据库、HGMD突变数据库比对突变位点,根据SIFT、Polyphen、MutationTaster、CADD数据库预测突变位点的致病性。结果: 25例患者中,发现抗凝因子相关基因PROC突变患者8例,PROS1突变患者2例,SERPINC1突变患者3例;凝血相关基因F5突变1例,F2突变1例,F8突变1例,CBS突变2例,MTHFR突变1例。结论: 应用第二代基因测序分析有助于诊断遗传性易栓症相关静脉血栓,指导精准治疗。

关键词: 静脉血栓形成/遗传学血栓形成倾向/遗传学血液凝固抗凝血酶类基因突变    
Abstract:

Objective: To screening coagulation and anticoagulation genes and mutations in Chinese patients with venous thromboembolism suspected of inhered thrombophilia. Methods: Patients with venous thromboembolism caused by inherited thrombophilia were collected from the second Affiliated Hospital, Zhejiang University School of Medicine during June 2015 and December 2017. Gene sequencing was conducted to screen the anticoagulant-related genes PROC, PROS1, SERPINC1 and coagulation-related genes F5, F2, F8, CBS, MTHFR. The thousands of human genome database, ESP6500 database, Genoma database, HGMD mutation database were used to compare the mutation sites, and the pathogenic mutation sites were predicted according to SIFT, Polyphen, MutationTaster and CADD databases. Results: Among 25 patients screened, eight patients with PROC mutations, two patients with PROS1 mutations, three patients with SERPINC1 mutations; one patient with F5 mutation, one patient with F2 mutation, one patient with F8 mutation, two patient with CBS mutations, and one patient with MTHFR mutation. Conclusion: The second generation gene sequencing helps to diagnose inherited thrombophilia and guide the decision of anticoagulation duration.

Key words: Venous thrombosis/genetics    Thrombophilia/genetics    Blood coagulation    Antithrombins    Genes    Mutation
收稿日期: 2018-08-14 出版日期: 2019-03-15
:  R446  
基金资助: 国家自然科学基金(81300236, 81670433)
通讯作者: 刘震杰     E-mail: yinli18c@zju.edu.cn;lawson4001@zju.edu.cn
作者简介: 尹黎(1990-), 女, 博士, 住院医师, 主要从事血管外科基础及临床研究; E-mail:yinli18c@zju.edu.cn; https://orcid.org/0000-0003-2008-9738
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引用本文:

尹黎,李歌,沈健,刘震杰. 遗传性易栓症筛查及相关基因检测分析[J]. 浙江大学学报(医学版), 2018, 47(6): 606-611.

YIN Li,LI Ge,SHEN Jian,LIU Zhenjie. Screening for inherited thrombophilia and genome sequencing. J Zhejiang Univ (Med Sci), 2018, 47(6): 606-611.

链接本文:

http://www.zjujournals.com/med/CN/10.3785/j.issn.1008-9292.2018.12.07        http://www.zjujournals.com/med/CN/Y2018/V47/I6/606

图 1  疑似遗传性易栓症患者的筛查流程
患者序号 性别 年龄 突变基因 突变位点 氨基酸改变 静脉血栓栓塞症类型
1 24 PROS1 g.78261A>G p.Y266C 深静脉血栓形成
2 24 PROS1 Ch1:93643083, C>T 剪切区域突变 深静脉血栓形成
PROC c.975C>G p.V325V
3 42 PROC c.631C>T p.R211W 深静脉血栓形成+肺栓塞
4 23 PROC c.962C>T p.P321L 深静脉血栓形成
5 21 PROC c.944C>T p.T340M 深静脉血栓形成
6 29 PROC c.267G>A p.G423S 深静脉血栓形成+肺栓塞
7 22 PROC c.889G>C p.D297H 深静脉血栓形成
8 45 PROC c.1152C>G p.N384K 深静脉血栓形成+肺栓塞
c.1207G>T p.G403W
9 25 PROC c.1273 G>A p.R393C 深静脉血栓形成
10 35 SERPINC1 c.1274G>A p.R425H 深静脉血栓形成
11 44 SERPINC1 c.967T>C p.S323P 深静脉血栓形成
12 26 SERPINC1 c.667T>C p.S223P 深静脉血栓形成+肺栓塞
表 1  抗凝因子基因突变患者的突变位点及类型
图 2  一例蛋白C缺乏症患者的遗传系谱及基因突变位点
患者序号 性别 年龄 突变基因 突变位点 氨基酸改变 静脉血栓栓塞症类型
2 24 F5 c.1154A>G p.N385S 深静脉血栓形成
CBS Ch21:44480542 G>A 剪切区域突变
9 25 F8 c.3169G>C p.E1057K 深静脉血栓形成
13 32 F2 c.1621C>T p.R541W 深静脉血栓形成
14 29 CBS c.400G>A p.G134R 深静脉血栓形成+肺栓塞
15 24 MTHFR chr1:11862935 T>C 剪切区域突变 深静脉血栓形成
表 2  凝血因子基因突变患者的突变位点及类型
1 LIJFERING W M , ROSENDAAL F R , CANNEGIETER S C . Risk factors for venous thrombosis-current understanding from an epidemiological point of view[J]. Br J Haematol, 149 (6): 824- 833
doi: 10.1111/bjh.2010.149.issue-6
2 BAGOT C N , ARYA R . Virchow and his triad:a question of attribution[J]. Br J Haematol, 2008, 143 (2): 180- 190
doi: 10.1111/bjh.2008.143.issue-2
3 MATEO J , OLIVER A , BORRELL M et al. Increased risk of venous thrombosis in carriers of natural anticoagulant deficiencies. Results of the family studies of the Spanish Multicenter Study on Thrombophilia (EMET study)[J]. Blood Coagul Fibrinolysis, 1998, 9 (1): 71- 78
doi: 10.1097/00001721-199801000-00009
4 MATEO J , OLIVER A , BORRELL M et al. Laboratory evaluation and clinical characteristics of 2, 132 consecutive unselected patients with venous thromboembolism——results of the Spanish Multicentric Study on Thrombophilia (EMET-Study)[J]. Thromb Haemost, 1997, 77 (3): 444- 451
doi: 10.1055/s-0038-1655986
5 BERTINA R M . Genetic approach to thrombophilia[J]. Thromb Haemost, 2001, 86 (1): 92- 103
6 DOUKETIS J , AGENO W , CARRIER M et al. Managing challenging patients with venous thromboembolism:a practical, case-based approach[J]. Pol Arch Intern Med, 2017, 127 (1): 41- 46
7 WEITZ J I , JAFFER I H . Optimizing the safety of treatment for venous thromboembolism in the era of direct oral anticoagulants[J]. Pol Arch Med Wewn, 2016, 126 (9): 688- 696
8 CROWTHER M A , KELTON J G . Congenital thrombophilic states associated with venous thrombosis:a qualitative overview and proposed classification system[J]. Ann Intern Med, 2003, 138 (2): 128- 134
doi: 10.7326/0003-4819-138-2-200301210-00014
9 TAIT R C , WALKER I D , PERRY D J et al. Prevalence of antithrombin deficiency in the healthy population[J]. Br J Haematol, 1994, 87 (1): 106- 112
doi: 10.1111/j.1365-2141.1994.tb04878.x
10 WHITLATCH N L , ORTEL T L . Thrombophilias:when should we test and how does it help?[J]. Semin Respir Crit Care Med, 2008, 29 (1): 25- 39
11 GARCíA DE FRUTOS P , FUENTES-PRIOR P , HURTADO B et al. Molecular basis of protein S deficiency[J]. Thromb Haemost, 2007, 98 (3): 543- 556
12 MARGAGLIONE M , BRANCACCIO V , GIULIANI N et al. Increased risk for venous thrombosis in carriers of the prothrombin G-->A 20210 gene variant[J]. Ann Intern Med, 1998, 129 (2): 89- 93
doi: 10.7326/0003-4819-129-2-199807150-00003
13 GU Y , SHEN W , ZHANG L et al. Deficiency of antithrombin and protein C gene in 202 Chinese venous thromboembolism patients[J]. Int J Lab Hematol, 2014, 36 (2): 151- 155
doi: 10.1111/ijlh.12146
14 WANG D , CUI G , HU S et al. Subtypes of SERPINC1 mutations and the thrombotic phenotype of inherited antithrombin deficient individuals in Chinese Han population[J]. Blood Cells Mol Dis, 2016, 62:38- 41
doi: 10.1016/j.bcmd.2016.10.029
15 MILLAR D S , JOHANSEN B , BERNTORP E et al. Molecular genetic analysis of severe protein C deficiency[J]. Hum Genet, 2000, 106 (6): 646- 653
16 BROEKMANS A W . Hereditary protein C deficiency[J]. Haemostasis, 1985, 15 (4): 233- 240
17 COOPER P C , HILL M , MACLEAN R M . The phenotypic and genetic assessment of protein C deficiency[J]. Int J Lab Hematol, 2012, 34 (4): 336- 346
doi: 10.1111/ijlh.2012.34.issue-4
18 WYPASEK E , UNDAS A . Protein C and protein S deficiency-practical diagnostic issues[J]. Adv Clin Exp Med, 2013, 22 (4): 459- 467
19 WYPASEK E , CORRAL J , ALHENC-GELAS M et al. Genetic characterization of antithrombin, protein C, and protein S deficiencies in Polish patients[J]. Pol Arch Intern Med, 2017, 127 (7-8): 512- 523
20 CAMPBELL I A , BENTLEY D P , PRESCOTT R J et al. Anticoagulation for three versus six months in patients with deep vein thrombosis or pulmonary embolism, or both:randomised trial[J]. BMJ, 2007, 334 (7595): 674
doi: 10.1136/bmj.39098.583356.55
21 RIDKER P M , MILETICH J P , HENNEKENS C H et al. Ethnic distribution of factor V Leiden in 4047 men and women. Implications for venous thromboembolism screening[J]. JAMA, 1997, 277 (16): 1305- 1307
doi: 10.1001/jama.1997.03540400055031
22 CONNORS J M . Thrombophilia testing and venous thrombosis[J]. N Engl J Med, 2017, 377 (23): 2297- 2298
doi: 10.1056/NEJMc1713797
23 COUTURAUD F , LEROYER C , TROMEUR C et al. Factors that predict thrombosis in relatives of patients with venous thromboembolism[J]. Blood, 2014, 124 (13): 2124- 2130
doi: 10.1182/blood-2014-03-559757
24 VAN VLIJMEN E F , WIEWEL-VERSCHUEREN S , MONSTER T B et al. Combined oral contraceptives, thrombophilia and the risk of venous thromboembolism:a systematic review and meta-analysis[J]. J Thromb Haemost, 2016, 14 (7): 1393- 1403
doi: 10.1111/jth.13349
25 GERHARDT A , SCHARF R E , GREER I A et al. Hereditary risk factors for thrombophilia and probability of venous thromboembolism during pregnancy and the puerperium[J]. Blood, 2016, 128 (19): 2343- 2349
doi: 10.1182/blood-2016-03-703728
26 KEARON C , AKL E A , ORNELAS J et al. Antithrombotic therapy for VTE disease:CHEST guideline and expert panel report[J]. Chest, 2016, 149 (2): 315- 352
doi: 10.1016/j.chest.2015.11.026
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