ASSOCIATION BETWEEN SPECIFIC THROMBOPHILIA- RELATED GENE POLYMORPHISMS AND RECURRENT PREGNANCY LOSS IN EGYPTIAN COHORT
Abstract
Recurrent pregnancy loss (RPL) affects an estimated 1–5% of couples worldwide, and inherited thrombophilia is implicated in up to 40% of unexplained cases. This study investigated the association between unexplained RPL and three thrombophilia-related gene polymorphisms — Factor V Leiden (FVL) 1691G>A, β-fibrinogen -455G>A, and plasminogen activator inhibitor-1 (PAI-1) 4G/5G — in an Egyptian cohort. This case-control study enrolled 200 women at Shebin El-Kom Teaching Hospital: 100 with unexplained RPL (two or more miscarriages) and 100 controls with no history of pregnancy loss. Genotyping was performed by multiplex PCR and reverse-hybridization strip assay (CVD StripAssay®). Genotype and allele frequencies were compared between groups using Fisher’s exact test and logistic regression, with Hardy-Weinberg equilibrium assessed in controls. The GG genotype and G allele of FVL 1691G>A were significantly more frequent in the RPL group than in controls (47% vs. 31%, P = 0.048; 66.5% vs. 54%, P = 0.011). The 4G/4G genotype and 4G allele of PAI-1 4G/5G were also significantly more frequent among women with RPL (49% vs. 31%, P = 0.030; 68.5% vs. 56%, P = 0.009). No significant association was found between the β-fibrinogen -455G>A polymorphism and RPL (P > 0.05 for all comparisons). FVL 1691G>A and PAI-1 4G/5G polymorphisms are significantly associated with unexplained RPL in this Egyptian cohort, while β-fibrinogen - 455G>A is not. These findings support incorporating FVL and PAI-1 genotyping into an expanded thrombophilia screening panel for women with unexplained RPL.
References
Al-Astal M. G. and Sharif F. A.(2014). Beta-fibrinogen (-455 G/A) and integrin beta-3 (PLA1/A2) polymorphisms and recurrent pregnancy loss in Gaza Strip-Palestine. Int. J. Reprod Contracept Obstet Gynecol 3:134-8.
Attili R., Hussein A., Odeh H. and Hejaz H. (2019). Prevalence of thrombophilia in Palestine and the association of thrombophilic gene polymorphisms with recurrent pregnancy loss. Res. J. Obstet Gynecol.12:6-10.
Balogh I., Szôke G., Kárpáti L., Wartiovaara U., Katona E., Komáromi I., Haramura G., Pfliegler G., Mikkola H. and Muszbek L. (2000). Val34Leu polymorphism of plasma factor XIII:biochemistry and epidemiology in familial thrombophilia. Blood, 96(7): 2479-86.
Board P. G., Losowsky M. S. and Miloszewski K. J. (1993). Factor XIII: inherited and acquired deficiency. Blood Rev. 7: 229-42.
Brenner B., Hoffman R., Blumenfeld Z., Weiner Z. and Younis J. S. (2000). Gestational outcome in thrombophilic women with recurrent pregnancy loss treated by enoxaparin. Thromb Haemost; 83:693-7.
Brenner B., Sarig G., Weiner Z., Younis J., Blumenfeld Z. and Lanir N. (1999). Thrombophilic polymerphisms are common in women with fetal loss without apparent cause. Thromb Haemost; 82:69.
Brown H. L. (1991). Antiphospholipid antibodies and recurrent pregnancy loss. Clin. Obstet Gynecol; 34:17-26.
Carp H., Salomon O., Seidman D., Dardik R., Rosenberg N. and Inbal A. (2002). Prevalence of genetic markers for thrombo-philia in recurrent pregnancy loss. Hum. Reprod; 17:1633-7.
Chen H., Nio S. and Lu M. (2015). Association between Plasminogen Activator Inhibitor-1 Gene Polymorphisms and Recurrent Pregnancy Loss: A Systematic Review and Meta-Analysis. Am. J. Reprod Immun.; 73: 292-300.
Djordevic V., Gvozdenov M., Pruner I., Kovac M. and Tomic B. et al. (2013). The prevalence of PAI-1 4G/5G polymorphism in women with fetal loss - first data for a Serbian population. Journal of Medical Biochemistry; 203-7.
Dossenbach-Glaninger A., van Trotsenburg M., Oberkanins C. and Atamaniuk J. (2013). Risk for Early Pregnancy Loss by Factor XIII Val34Leu: The Impact of Fibrinogen Concentration. Journal of Clinical Laboratory Analysis ; 27: 444-9.
ESHRE (2018). Guideline Group on RPL, Bender Atik R, Christiansen OB, et al. ESHRE guideline: recurrent pregnancy loss. Hum Reprod Open.; (2):hoy 004. doi:10.1093/hropen/hoy004.
Farahmand K., Totonchi M., Hashemi M., Reyhani Sabet F., Kalantari H., Gourabi H., et al. (2016). Thrombophilic gene alterations as risk factor for recurrent pregnancy loss. J. Matern Fetal Neonatal Med; 29:1269-1273.
Girolami A., Scandellari R., Lombardi A. M., Girolami B., Bortoletto E. and Zanon E. (2005). Pregnancy and oral contraceptives in factor V deficiency: a study of 22 patients (five homozygotes and 17 heterozygotes) and review of the literature. Haemophilia; 11:26-30.
Grandone E., Margaglione M., Colaizzo D., d’Addedda M., Cappucci G., Vecchione G., Sciannamé N., Pavone G. and Di Minno G. (1997). Factor V Leiden is associated with repeated and recurrent unexplained fetal losses. Thromb Haemost; 77:822-4.
Hohlagschwandtner M., Unfried G., Heinze G., Huber J. C., Nagele F. and Tempfer C. (2003). Combined thrombophilic polymorphisms in women with idiopathic recurrent miscarriage. Fertil Steril; 79:1141-8.
Huang Z., Tang W., Liang Z., Chen Q., Li M., Li Y., Lao S., Pan H. and Huang L, et al. (2017). Plasminogen activator inhibitor-1 polymorphism confers a genetic contribution to the risk of recurrent spontaneous abortion: an updated meta-analysis. Reprod Sci.;24:1551-60.
Ivanov P. D., Komsa-Penkova R. S., Konova E. I., Kovacheva K. S., Simeonova M. N. and Popov J. D. (2009). Association of inherited thrombophilia with embryonic and postembryonic recurrent pregnancy loss. Blood Coagul Fibrinolysis; 2:134-40.
Karami F., Askari M. and Modarressi M. H. (2018). Investigating Association of rs5918 Human Platelets Antigen 1 and rs1800790 Fibrinogen β Chain as Critical Players with Recurrent Pregnancy Loss. Med. Sci. (Basel).;6:98.
Khosravi F., Zarei S., Ahmadvand N., Akbarzadeh-Pasha Z., Savadi E., Zarnani A. H., Sadeghi M. R. and Jeddi-Tehrani M. (2014). Association between plasminogen activator inhibitor-1 gene polymorphism and different subgroups of recurrent miscarriage and implantation failure. J., Assist Reprod Genet.; 31:121-4.
Kreutz R. P., Bitar A., Owens J., Desta Z., Breall J. A., von der Lohe E., Sinha A., Vatta M., Nystrom P., Jin Y. and Flockhart D. A. (2014). Factor XIII Val34Leu polymorphism and recurrent myocardial infarction in patients with coronary artery disease. J. Thromb Thrombolysis 38: 380-7.
Levin, Veronika, Rachel Booth and Shahab (2020). Minassian. Bleeding Disorders and ART. In: Textbook of Assisted Reproduction. Springer;.
Li J, Wu H., Chen Y., Wu H., Xu H. and Li L. (2015)Genetic association between FXIII and β-fibrinogen genes and women with recurrent spontaneous abortion: A meta-analysis. J. Assist Reprod Genet.;32:817-25.
Martinelli I., Sacchi E., Landi G., Taioli E., Duc F. and Mannucci P. M. (1998). High risk of cerebral-vein thrombosis in carriers of a prothrombin-gene mutation and in users of oral contraceptives. N Engl J. Med; 338:1793-7.
McNamee K., Dawood F., Farquharson R., (2012). Recurrent miscarriage and thrombophilia: An update. Curr Opin Obstet Gynecol; 24:229-34.
Mikkola H. and Palotie A. (1996). Gene defects in congenital factor XIII deficiency. Semin Thromb Hemost 1996; 22: 393-8.
Practice Committee of the American Society for Reproductive Medicine. (2013). Definitions of infertility and recurrent pregnancy loss: a committee opinion. Fertil Steril.;99:63.
RCOG. (2019). Recurrent Miscarriage, Investigation and Treatment of Couples (Green-top Guideline No. 17) 19/05/2011. Available from: https://www.rcog.org.uk/en/guidelines-research-services/guidelines/gtg17/. [last accessed 2019 Dec 5].
Rey E., Kahn S. R., David M. and Shrier I. (2003). Thrombophilic disorders and fetal loss: a meta-analysis. Lancet; 361:901-8.
Reznikoff-Etiévan M. F., Cayol V., Carbonne B., Robert A, Coulet F. and Milliez J. (2001). Factor V Leiden and G20210A prothrombin mutations are risk factors for very early recurrent miscarriage. BJOG; 108:1251-4.
Ridker, C. H., Klaus Lindpaintner, Stampfer, Eisenberg P. R. and Joseph P. Miletich (1995). Mutation in the gene coding for coagulation factor V and the risk of myocardial infarction, stroke, and venous thrombosis in apparently healthy men. N. Engl. J. Med.; 332(14):912-7.
Robertson L., Wu. O., Langhorne P., Twaddle S., Clark P., Lowe G. D., Walker I. D., Greaves M., Brenkel I., Regan L. and Greer I. A. (2006). Thrombophilia in pregnancy: a systematic review. Br J Haematol; 132:171-96.
Rodeghiero F. and Tosetto A. (19990. Activated protein C resistance and factor V Leiden mutation are independent risk factors for venous thromboembolism. Ann. Intern. Med.;130(8):643-50.
Rosendaal F. R., Koster T., Vandenbroucke J. P. and Reitsma P. H. (1995). High risk of thrombosis in patients homozygous for factor V Leiden (activated protein C resistance). Blood 1995; 85:1504-8.
Sarig G., Younis J. S., Hoffman R., Lanir R., Blumenfeld Z. and Brenner B. (2002). Thrombophilia is common in women with idiopathic pregnancy loss and is associated with late pregnancy wastage. Fertil Steril; 77:342-7.
Shi X., Xie X., Jia Y. and Li S. (2017). Maternal genetic polymorphisms and unexplained recurrent miscarriage: a systematic review and meta-analysis. Clin. Genet.; 91:265-84.
Su M. T., Lin S. H., Chen Y. C. and Kuo P. L. (2013). Genetic association studies of ACE and PAI-1 genes in women with recurrent pregnancy loss. Thromb Haemost.;109(8):15.
Ticconi C., Mancinelli F., Gravina P., Federici G., Piccione E. and Bernardini S. (2011). Beta-fibrinogen G-455A polymorphisms and recurrent miscarriage. Gynecol Obstet Invest.; 71:198-201.
Torabi R., Zarei S., Zeraati H., Zarnani A. H., Akhondi M. M. and Hadavi R. (2012). Combination of thrombophilic gene polymorphisms as a cause of an increased risk of recurrent pregnancy loss. J. Reprod Infertil;13:89-94.
Tybjaerg-Hansen A., Agerholm-Larsen B., Humphries S. E., Abildgaard S., Schnohr P. and Nordestgaard B. G. A. (1997). Common polymorphism (G-455-->A) in the beta-fibrinogen promoter is an independent predictor of plasma fibrinogen, but not of ischemic heart disease. A study of 9,127 individuals based on the Copenhagen City Heart Study. J. Clin Invest.;99:3034-9. doi: 10.1172/JCI119499.
Younis J. S., Ohel G., Brenner B. and Ben-Ami M. (1997). Familial thrombophilia - the scientific rationale for thromboprophylaxis in recurrent pregnancy loss? Hum. Reprod; 12:1389-90.
Younis J. S., Ohel G., Brenner B., Haddad S., Lanir N. and Ben-Ami M. (2000.). The effect of thromboprophylaxis on pregnancy outcome in patients with recurrent pregnancy loss associated with factor V Leiden polymorphism. BJOG; 107:415-9.
Yousefian E., Kardi M. T. and Allahveisi A. (2014). Methylen-etetrahydrofolate reductase C677T and A1298C polymorphism in Iranian women with idiopathic recurrent pregnancy losses. Iran Red Crescent Med J.; 16:e16763.
Zhao L., Bracken M., DeWan A. and Chen A. (2013). Association between the SERPINE1(PAI-1) 4G/5G insertion/deletion promoter polymorphism (rs1799889) and pre-eclampsia: a systematic review and meta-analysis. Molecular Human Reproduction; 19(3): 136– 43.
