ANALYSIS OF FACTOR V AND MTHFR GENES AS RISK FACTORS CONSTITUTING SUSCEPTIBILITY TO NEURAL TUBE DEFECTS: A CASE-CONTROL STUDY FROM TURKEY

Yıl 2020, Cilt: 22 Sayı: 1, 105 - 112, 30.04.2020

Kağan Kamaşak Zeynep Yeğin İbrahim Yıldırım

https://doi.org/10.24938/kutfd.676442

Öz

Objective: This study targeted to bring a molecular perspective to occult neural tube defects and thus develop future preventive personalized medicine strategies. The roles of three genetic variations namely Factor V Leiden (FVL) (rs6025), MTHFR A1298C (rs1801131), and MTHFR C677T (rs1801133) were investigated in a Turkish cohort including both the mothers and children to better analyze the potential inherited effects of these variations.

Material and Methods: Children affected with neural tube defects (NTDs) and control children without NTDs were included in the study together with their mothers. DNA extractions were performed from collected blood samples with standard salting-out procedure. Isolated DNAs were genotyped with Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method.

Results: In terms of Factor V Leiden (FVL) (rs6025) in NTD risk, there was no statistically important association between mothers with NTDs-affected children and control group mothers (p=0.639). However, a statistically significant association was observed when NTDs-affected children were compared with the ones not affected (p=0.0144). There was a statistically important association of MTHFR A1298C (rs1801131) both in mothers comparisons and comparison between NTDs-affected children and not-affected children (respectively, p=0.005; 0.008). MTHFR C677T (rs1801133) genotypes and/or alleles did not act as risk factors for NTD development neither in mothers nor in children in this study (p˃0.05).

Conclusion: Our study indicates FVL mutation as an increased risk factor for NTD development, independently from the genotypes of mothers. MTHFR A1298C (rs1801131) homozygous AA genotype and A allele elevated the risk of NTD development both in mothers and children referring to the inherited role of this variation in Turkish population. However, MTHFR C677T (rs1801133) variation can not be considered as a risk factor in NTD development in our population.

Anahtar Kelimeler

Neural tube defects (NTDs), genetic association, risk factors, , Factor V Leiden, 5-10-methylenetetrahydrofolate reductase (MTHFR)

Proje Numarası

DUBAP 11-TF-62

Nöral Tüp Defektlerinde Duyarlılık Oluşturan Risk Faktörleri Olarak Faktör V ve MTHFR Genlerinin Analizi: Türkiye'den Bir Vaka-Kontrol Çalışması

Öz

Amaç: Bu çalışma konjenital olmayan nöral tüp defektlerine moleküler bir perspektif sunmayı ve geleceğe yönelik önleyici bireyselleştirilmiş tıp stratejileri geliştirmeyi hedeflemiştir. Üç genetik varyasyon; Factor V Leiden (FVL) (rs6025), MTHFR A1298C (rs1801131) ve MTHFR C677T (rs1801133) bu varyasyonların potansiyel kalıtımsal etkilerini daha iyi analiz etmek için hem anneleri hem de çocukları içeren bir Türk kohortunda araştırılmıştır.

Gereç ve Yöntemler: Nöral tüp defektli (NTD) çocuklar ve NTD'li olmayan kontrol grubu çocukları anneleriyle beraber çalışmaya dahil edilmiştir. Toplanılan kan örneklerinden standart tuzla çöktürme prosedürüyle DNA ekstraksiyonları gerçekleştirilmiştir. İzole edilen DNA'lar Polimeraz Zincir Reaksiyonu-Restriksiyon Fragment Uzunluk Polimorfizmi (PCR-RFLP) metoduyla genotiplendirilmiştir.

Bulgular: Factor V Leiden (FVL) (rs6025) mutasyonunun NTD riskini arttırması açısından NTD'li çocuğa sahip annelerle kontrol grubu anneleri arasında istatistiksel olarak önemli bir ilişki mevcut değildir (p=0.639). Ancak, NTD'li çocuklarla etkilenmemiş çocuklar karşılaştırıldığı zaman istatistiksel olarak önemli bir ilişki gözlenmiştir (p=0.0144). MTHFR A1298C (rs1801131) açısından ise hem annelerin kendi aralarındaki karşılaştırmada hem de NTD'li çocuklar ve etkilenmemiş çocuklar arasında istatistiksel olarak önemli bir ilişki mevcuttur (sırasıyla p=0.005; 0.008). MTHFR C677T (rs1801133) genotipleri ve/veya allelleri ise bu çalışmada ne annelerde ne de çocuklarda NTD gelişimi açısından risk faktörü olarak rol oynamadığı görülmüştür (p˃0.05).

Sonuç: Çalışmamız, NTD gelişimi açısından FVL mutasyonunun annelerin genotiplerinden bağımsız olarak artmış bir risk faktörü olduğuna işaret etmektedir. MTHFR A1298C (rs1801131) homozigot AA genotipi ve A alleli bu varyasyonun Türk popülasyonunda kalıtımsal etkisine de vurgu yapacak şekilde hem annelerde hem de çocuklarda NTD gelişimi riskinde artışa yol açmaktadır. Ancak, MTHFR C677T (rs1801133) varyasyonu populasyonumuzda NTD gelişimi açısından bir risk faktörü olarak dikkate alınamamaktadır.

Anahtar Kelimeler

Factor V Leiden, Nöral tüp defektleri, genetik assosiasyon, risk faktörleri, 5-10-methylenetetrahydrofolate reductase (MTHFR)

Destekleyen Kurum

DİCLE ÜNİVERSİTESİ BİLİMSEL ARAŞTIRMA PROJELERİ KOORDİNATÖRLÜĞÜ

Proje Numarası

DUBAP 11-TF-62

Teşekkür

Bu çalışma, Dicle Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğünce DUBAP 11-TF-62 numarası ile desteklenmiştir.

Kaynakça

• 1. Paul S, Sadhukhan S, Munian D, Bankura B. Association of FOLH1, DHFR, and MTHFR gene polymorphisms with susceptibility of neural tube defects: A case control study from Eastern India. Birth Defects Research. 2018;110(14):1129-38.
• 2. Cai CQ, Fang YL, Shu JB, Zhao LS, Zhang RP, Cao LR et al. Association of neural tube defects with maternal alterations and genetic polymorphisms in one-carbon metabolic pathway. Ital J Pediatr. 2019;45(1):37.
• 3. Liu J, Zhang Y, Jin L, Li G, Wang L, Bao Y et al. Variants in maternal COMT and MTHFR genes and risk of neural tube defects in offspring. Metab Brain Dis. 2015;30(2):507-13.
• 4. Fang Y, Zhang R, Zhi X, Zhao L, Cao L, Wang Y et al. Association of main folate metabolic pathway gene polymorphisms with neural tube defects in Han population of Northern China. Childs Nerv Syst. 2018;34(4):725-9.
• 5. Elwood JM, Little J, Elwood JH. Epidemiology and control of neural tube defects. 1st ed. Oxford (UK). Oxford University Press, 1992.
• 6. Centers for Disease Control and Prevention. Recommendations for the use of folic acid to reduce the number of cases of spina bifida and other neural tube defects. MMWRRecomm Rep. 1992;41(RR-14):1-7.
• 7. Helgadottir LB, Skjeldestad FE, Jacobsen AF, Sandset PM, Jacobsen EM. The association of inherited thrombophilia and intrauterine fetal death: a case-control study. Blood Coagul Fibrinolysis. 2011;22(8):651-6.
• 8. Goyette P, Sumner JS, Milos R, Duncan AM, Rosenblatt DS, Matthews RG et al. Human methylenetetrahydrofolate reductase: isolation of cDNA mapping and mutation identification. Nat Genet. 1994;7(2):195-200.
• 9. Mahmutbegović E, Marjanović D, Medjedović E, Mahmutbegović N, Dogan S, Valjevac A et al. Prevalence of F5 1691G>A, F2 20210G>A, and MTHFR 677C>T polymorphisms in Bosnian women with pregnancy loss. Bosn J Basic Med Sci. 2017;17(4):309-14.
• 10. Ornstein DL and Cushman M. Cardiology patient page. Factor-V Leiden. Circulation. 2003;107(15):e94-7.
• 11. Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988;16(3):1215.
• 12. Beauchamp NJ, Daly ME, Hampton KK, Cooper PC, Preston E. High prevalence of a mutation in the factor V gene within the UK population: relationship to activated protein C resistance and familial thrombosis. Hypertension. 1994;88(1):219-22.
• 13. Goyette P, Frosst P, Rosenblatt DS, Rozen R. Seven novel mutations in the methylenetetrahydrofolate reductase gene and genotype/phenotype correlations in severe methylenetetrahydrofolate reductase deficiency. Am J Hum Genet. 1995;56(5):1052-9.
• 14. Van der Put NM, Gabreels F, Stevens EM, Smeitink JA, Trijbels FJ, Eskes TK et al. A second mutation in the MTHFR gene: an additional risk factor for neural-tube defects? Am J Hum Genet. 1998;62(5):1044 -51.
• 15. Bertina RM, Koeleman BP, Koster T, Rosendaal FR, Dirven RJ, de Ronde H et al. Mutation in blood coagulation factor-V associated with resistance to activated protein C. Nature. 1994;369(6475):64-7.
• 16. Ibrahim-Kosta M, Suchon P, Couturaud F, Smadja D, Olaso R, Germain M et al. Minor allele of the factor-V K858R variant protects from venous thrombosis only in non-carriers of factor-V Leiden mutation. Sci Rep. 2019;9(1):3750.
• 17. Yalcintepe S, Ozdemir O, Hacivelioglu SO, Akurut C, Koc E, Uludag A et al. Multiple Inherited Thrombophilic Gene Polymorphisms in Spontaneous Abortions in Turkish Population. Int J Mol Cell Med. 2015;4(2):120-7.
• 18. Udry S, Aranda FM, Latino JO, de Larrañaga GF. Paternal factor V Leiden and recurrent pregnancy loss: a new concept behind fetal genetics? Reply. J Thromb Haemost. 2014;12(10):1759-60.
• 19. Ahmed NA, Adam I, Elzaki SEG, Awooda HA, Hamdan HZ. Factor-V Leiden G1691A and prothrombin G20210A polymorphisms in Sudanese women with preeclampsia, a case -control study. BMC Med Genet. 2019;20(1):2.
• 20. Dudding T, Heron J, Thakkinstian A, Nurk E, Golding J, Pembrey M et al. Factor V Leiden is associated with pre-eclampsia but not with fetal growth restriction: a genetic association study and meta-analysis. J Thromb Haemost. 2008;6(11):1869-75.
• 21. Dissanayake VH, Sirisena ND, Weerasekera LY, Gammulla CG, Seneviratne HR, Jayasekara RW. Candidate gene study of genetic thrombophilic polymorphisms in pre-eclampsia and recurrent pregnancy loss in Sinhalese women. J Obstet Gynaecol Res. 2012;38(9):1168-76.
• 22. Said JM, Brennecke SP, Moses EK, Walker SP, Borg AJ, Williams JT et al. Ethnic differences in the prevalence of inherited thrombophilic polymorphisms in an asymptomatic Australian prenatal population. Hum Biol. 2006;78(4):403-12.
• 23. Ryckman KK, Dagle JM, Kelsey K, Momany AM, Murray JC. Replication of genetic associations in the inflamation, complement, and coagulation pathways with intraventricular hemorrhage in LBW preterm neonates. Pediatr Res. 2011;70(1):90-5.
• 24. Aydin H, Arisoy R, Karaman A, Erdoğdu E, Çetinkaya A, B Geçkinli B et al. Evaluation of maternal serum folate, vitamin B12, and homocysteine levels and factor V Leiden, factor II g.20210G>A, and MTHFR variations in prenatally diagnosed neural tube defects. Turk J Med Sci. 2016;46(2):489-94.
• 25. Yu Y, Wang F, Bao Y, Lu X, Quan L, Lu P. Association between MTHFR gene polymorphism and NTDs in Chinese Han population. Int J Clin Exp Med. 2014;7(9):2901-6.
• 26. Dutta HK, Borbora D, Baruah M, Narain K. Evidence of gene-gene interactions between MTHFD1 and MTHFR in relation to anterior encephalocele susceptibility in Northeast India. Birth Defects Res. 2017;109(6):432-44.
• 27. Etheredge AJ, Finnell RH, Carmichael SL, Lammer EJ, Zhu H, Mitchell LE et al. Maternal and infant gene-folate interactions and the risk of neural tube defects. Am J Med Genet A. 2012;158A(10):2439-46.
• 28. Boyles AL, Billups AV, Deak KL, Siegel DG, Mehltretter L, Slifer SH et al. Neural tube defects and folate pathway genes: family-based association tests of gene-gene and gene-environment interactions. Environ Health Perspect. 2006;114(10):1547-52.