Phenotype and genotype diagnosis of 6 pedigrees with inherited coagulation factor Ⅹ deficiency

Abstract

Abstract:

Objective:To investigate the molecular pathogenesis of 6 pedigrees with inherited coagulation factorⅩ (FⅩ) deficiency and to assess the bleeding risk in patients with inherited FⅩ deficiency by thrombin generation test (TGT).Methods:Clinical data of the 6 pedigrees were collected. The FⅩ coagulation activity (FⅩ:C) was tested by clotting test and FⅩ antigen (FⅩ:Ag) was detected by double antibody sandwich enzyme-linked immunosorbent assay（ELISA）. The concentration of FⅩ in plasma was measured with Western blotting. The FⅩ gene (F10) mutation was analyzed by direct sequencing. Thrombin generation (TG) in six patients and some family members was measured using TET.Results:TenF10mutations were identified in 6 probands, five of which (p.Cys 246 Ser, p.Tyr 319 Cys, p.Leu 252 Phe, p.Arg 313 Gln and p. IVS5-2A>G) were novel and the other five(p.Phe71Ser, p.Val 338 Met, p.Gly 406 Ser, p.Gly 365 Ser and p.Cys 151 Arg) had been previously reported. TGT showed that endogenous thrombin potential (ETP) and peak height (Peak) had certain correlation with bleeding severity of FX-deficient patients.Conclusions:Five noval mutations of FⅩ are identified. Five patients were resulted from compound heterozygousF10mutations, while the remaining one is caused by one heterozygousF10mutation. TGT parameters including ETP and Peak could be used to assess the bleeding risk in patients with FⅩ deficiency, and TGT might serve as a useful laboratory tool to monitor the risk of bleeding.

Key words:Coagulation factor Ⅹ deficiency,Thrombin generation test,Phenotype,Genotype

CLC Number:

R446

LIU Jiajie, SHAO Yanyan, LIANG Qian, DING Qiulan. Phenotype and genotype diagnosis of 6 pedigrees with inherited coagulation factor Ⅹ deficiency[J]. Journal of Diagnostics Concepts & Practice, 2020, 19(1): 20-27.

Figures/Tables7

先证者	性别/ 年龄（岁）	临床表现	出血评分(分)	APTT (s)	PT (s)	FⅩ∶C			FⅩ∶ Ag	分型	基因检测
先证者	性别/ 年龄（岁）	临床表现	出血评分(分)	APTT (s)	PT (s)	APTT途径	PT途径	RVV途径	FⅩ∶ Ag	分型	基因检测
1	男/10	皮肤淤斑,换牙时出血, 肌肉出血	7	74.1	28.9	2.4	2.6	2.4	7.3	Ⅰ型	c.212T>C, p.Phe71Ser; c.1012G>A,p.Val338Met
2	女/79	月经量多,手术后出血难止,拔牙后出血	4	42.7	15.8	38.9	42.2	44.4	83.1	Ⅱ型	c.1216G>A, p.Gly406Ser
3	女/36	月经量多,外伤后出血难止	6	48.3	20.2	15.8	16.8	12.3	67.6	Ⅱ型	c.736T>A, p.Cys246Ser; c.956A>G, p.Tyr319Cys
4	女/24	自发鼻出血,皮肤易发淤斑,牙龈出血	4	55.9	26.5	4.2	7.2	6.2	35.4	Ⅱ型	c.1093G>A, p.Gly365Arg; c.754C>T, p.Leu252Phe
5	男/13	无出血症状	0	38.8	16.3	27.9	12.7	16.7	18.2	Ⅰ型	c.938G>A, p.Arg313Gln; IVS5-2A>G
6	男/26	鼻出血,牙龈出血,皮肤易发淤斑,关节出血,肌肉血肿	10	52.5	30.6	3.8	2.4	2.5	5.6	Ⅰ型	c.212T>C, p.Phe71Ser; c.451T>C, p.Cys151Arg

患者及家系成员	LT(min)	ETP (nmol*min)	峰值(nmol)	TTP(min)
P1-(Ⅱ-1)	13.33	98.23	3.77	28.67
P1-(Ⅰ-1)	7.00	408.12	41.61	12.67
P1-(Ⅰ-2)	6.76	626.77	33.18	17.12
P2-(Ⅱ-1)	7.83	763.50	55.81	15.17
P3-(Ⅱ-1)	10.33	224.32	15.60	17.67
P3-(Ⅲ-1)	9.00	944.50	88.70	14.67
P4-(Ⅱ-1)	30.32	99.90	5.04	40.84
P5-(Ⅱ-1)	7.83	295.56	26.57	13.83
P5-(Ⅰ-1)	6.00	764.93	68.50	12.00
P6-(Ⅱ-1)	5.17	83.91	3.04	14.67
正常对照者	6.17	969.31	92.49	11.67

References20

[1]	Hertzberg M. Biochemistry of factor Ⅹ[J]. Blood Rev, 1994, 8(1):56-62. pmid:8205010
[2]	Jackson CM. Factor Ⅹ[J]. Prog Hemost Thromb, 1984, 7:55-109. pmid:6397768
[3]	Uprichard J, Perry DJ. Factor Ⅹ deficiency[J]. Blood Rev, 2002, 16(2):97-110. pmid:12127953
[4]	Girolami A, Cosi E, Santarossa C, et al. Factor Ⅹ friuli coagulation disorder: almost 50 years later[J]. Clin Appl Thromb Hemost, 2018, 24(1):33-40. doi:10.1177/1076029616686423 URL
[5]	Menegatti M, Peyvandi F. Factor Ⅹ deficiency[J]. Semin Thromb Hemost, 2009, 35(4):407-415. doi:10.1055/s-0029-1225763pmid:19598069
[6]	Dorgalaleh A, Zaker F, Tabibian S, et al. Spectrum of factor Ⅹ gene mutations in Iranian patients with congenital factor Ⅹ deficiency[J]. Blood Coagul Fibrinolysis, 2016, 27(3):324-327. doi:10.1097/MBC.0000000000000435 URL
[7]	Brown DL, Kouides PA. Diagnosis and treatment of inherited factor Ⅹ deficiency[J]. Haemophilia, 2008, 14(6):1176-1182. doi:10.1111/j.1365-2516.2008.01856.xpmid:19141158
[8]	Peyvandi F, Menegatti M, Santagostino E, et al. Gene mutations and three-dimensional structural analysis in 13 families with severe factor Ⅹ deficiency[J]. Br J Haematol, 2002, 117(3):685-692. doi:10.1046/j.1365-2141.2002.03486.x URL
[9]	Girolami A, Vettore S, Scarparo P, et al. Persistent vali-dity of a classification of congenital factor Ⅹ defects based on clotting, chromogenic and immunological assays even in the molecular biology era[J]. Haemophilia, 2011, 17(1):17-20. doi:10.1111/j.1365-2516.2010.02328.xpmid:20546029
[10]	Duarte RCF, Rios DRA, Rezende SM, et al. Standardization and evaluation of the performance of the thrombin generation test under hypo- and hypercoagulability conditions[J]. Hematol Transfus Cell Ther, 2019, 41(3):244-252.
[11]	Tripodi A. Thrombin generation assay and its application in the clinical laboratory[J]. Clin Chem, 2016, 62(5):699-707. doi:10.1373/clinchem.2015.248625 URL
[12]	Joly BS, Sudrié-Arnaud B, Barbay V, et al. Thrombin generation test as a marker for high risk venous thrombosis pregnancies[J]. J Thromb Thrombolysis, 2018, 45(1):114-121. doi:10.1007/s11239-017-1572-3 URL
[13]	Liang Q, Chen Q, Ding Q, et al. Six novel missense mutations causing factor Ⅹ deficiency and application of thrombin generation test[J]. Thromb Res, 2013, 131(6):554-9. doi:10.1016/j.thromres.2013.04.014pmid:23664564
[14]	The F10 gene mutation database[R/OL]. [2020-01-31]. http://www.hgmd.cf.ac.uk/ac/all.php.
[15]	Millar DS, Elliston L, Deex P, et al. Molecular analysis of the genotype-phenotype relationship in factor Ⅹ deficiency[J]. Hum Genet, 2000, 106(2):249-257. pmid:10746568
[16]	Borhany M, Buthiau D, Rousseau F, et al. Genotyping of five Pakistani patients with severe inherited factor Ⅹ deficiency: identification of two novel mutations[J]. Blood Coagul Fibrinolysis, 2018, 29(7):622-625. doi:10.1097/MBC.0000000000000764 URL
[17]	Isshiki I, Favier R, Moriki T, et al. Genetic analysis of hereditary factor Ⅹ deficiency in a French patient of Sri Lankan ancestry:in vitroexpression study identified Gly366Ser substitution as the molecular basis of the dysfunctional factor Ⅹ[J]. Blood Coagul Fibrinolysis, 2005, 16(1):9-16. doi:10.1097/00001721-200501000-00002 URL
[18]	Miyata T, Kojima T, Suzuki K, et al. Factor Ⅹ Nagoya 1 and Nagoya 2: a CRM- factor Ⅹ deficiency and a dysfunctional CRM+ factor Ⅹ deficiency characterized by substitution of Arg306 by Cys and of Gly366 by Ser, respectively[J]. Thromb Haemost, 1998, 79(3):486-490. doi:10.1055/s-0037-1614930 URL
[19]	Mota L, Shetty S, Idicula-Thomas S, et al. Molecular basis of factor Ⅹ deficiency cases from India[J]. Haemophilia, 2010, 16(4):693-697. doi:10.1111/j.1365-2516.2010.02213.xpmid:20331754
[20]	Herrmann FH, Auerswald G, Ruiz-Saez A, et al. Factor Ⅹ deficiency: clinical manifestation of 102 subjects from Europe and Latin America with mutations in the factor 10 gene[J]. Haemophilia, 2006, 12(5):479-489. pmid:16919077