Turkiye Klinikleri Journal of Medical Sciences

.: ORIGINAL RESEARCH
Hemofili B Moleküler Analizinde Türkiye Deneyimi: F9 Gen Mutasyon Spektrumu ve Genotip-Fenotip İlişkisi
Turkey Experience in Molecular Analysis of Hemophilia B: F9 Gene Mutation Spectrum and Genotype-Phenotype Correlation
Esra IŞIKa, Bilçağ AKGÜNa, Kaan KAVAKLIb, Melike SEZGİN EVİMc, Canan ALBAYRAKd, Gülen TÜYSÜZ KİNTRUPe, Fahri ŞAHİNf, Bülent ANTMENg, Ebru YILMAZ KESKİN h, Salih GÜLERc, Davut ALBAYRAKı, Osman Alphan KÜPESİZe, Hüseyin ONAYi, Ferda ÖZKINAYa,i, Tahir ATİKa
aEge Üniversitesi Tıp Fakültesi, Çocuk Genetik Hastalıkları BD, İzmir, TÜRKİYE
bEge Üniversitesi Tıp Fakültesi, Çocuk Hematoloji BD, İzmir, TÜRKİYE
çUludağ Üniversitesi Tıp Fakültesi, Çocuk Hematoloji BD, Bursa, TÜRKİYE
dOndokuz Mayıs Üniversitesi Tıp Fakültesi, Çocuk Hematoloji BD, Samsun, TÜRKİYE
eAkdeniz Üniversitesi Tıp Fakültesi, Çocuk Hematoloji BD, Antalya, TÜRKİYE
fEge Üniversitesi Tıp Fakültesi, Hematoloji BD, İzmir, TÜRKİYE
gAcıbadem Adana Hastanesi, Çocuk Hematoloji BD, Adana, TÜRKİYE
hSüleyman Demirel Üniversitesi Tıp Fakültesi, Çocuk Hematoloji BD, Isparta, TÜRKİYE
ıMedical Park Samsun Hastanesi, Çocuk Hematoloji BD, Samsun, TÜRKİYE
iEge Üniversitesi Tıp Fakültesi, Tıbbi Genetik ABD, İzmir, TÜRKİYE
Turkiye Klinikleri J Med Sci. 2020;40(3):334-41
doi: 10.5336/medsci.2020-75066
Article Language: TR
Full Text
ÖZET
 Amaç: Hemofili B (HB), yaklaşık 30.000 canlı erkek doğumda bir görülen, X'e bağlı kalıtsal kanama bozukluğudur. Klinik bulgular, koagülasyon faktörü 9'un aktivite düzeyine göre değişkenlik gösterir. Bugüne kadar koagülasyon faktörü 9'u kodlayan F9 geninde 1.200'den fazla varyant tanımlanmıştır. Varyantların yaklaşık %70'ini nokta mutasyonları oluşturur. Bu çalışmada, Türkiye'deki HB hastalarında F9 gen varyant dağılımının belirlenmesi ve yeni varyantlar tanımlayarak genotip-fenotip ilişkisine katkıda bulunulması amaçlanmıştır. Gereç ve Yöntemler: Türkiye'deki sekiz farklı merkezde HB tanısı ile takip edilen ve Ege Üniversitesi Tıp Fakültesinde Kasım 2018-Ocak 2020 tarihleri arasında moleküler analizi yapılmış 55 hasta çalışmaya alınmıştır. Olguların klinik ve laboratuvar bulguları hastane kayıtlarından elde edilmiştir. F9 geni dizi analizi, yeni nesil dizi analizi platformu (MiSeq' Illimuna) kullanılarak yapılmıştır. Saptanan yeni varyantların patojeniteleri ACMG 2015 kriterlerine göre sınıflandırılmıştır. Saptanan varyantlar ile fenotip ilişkisi değerlendirilmiştir. Bulgular: Çalışmaya alınan 55 erkek hastanın 33 (%60)'ünde ağır, 15 (%27,3)'inde orta ve 7 (%12,7)'sinde hafif HB fenotipi vardı. F9 dizi analizi sonucunda 54 (%98,2) olguda 46 farklı varyant saptandı. Bu varyantların 30 (%63,8)'u yanlış anlamlı, 9 (%19,1)'u anlamsız, 3 (%6,4)'ü çerçeve kayması ve 4 (%8,5)'ü kırpılma noktası mutasyonuydu. Belirlenen varyantların 10 tanesi daha önce literatürde tanımlanmamıştı. Sonuç: Bu çalışmada, Türkiye'deki HB hastalarında, moleküler analizin tanı başarısı ve F9 geninde varyant dağılımı literatüre benzer şekilde bulunmuştur. Çalışmanın sonuçları, HB'nin genotipik olarak heterojen bir hastalık olduğunu desteklemektedir. On yeni varyant ilk kez bu çalışma ile tanımlanmış ve hastalığın genotip-fenotip ilişkisine katkıda bulunulmuştur.

Anahtar Kelimeler: Yeni nesil dizi analizi; hemofili B; mutasyon; genotip-fenotip ilişkisi
ABSTRACT
 Objective: Hemophilia B (HB) is an X-linked hereditary bleeding disorder seen in approximately one in 30,000 live male births. Clinical findings vary according to the activity level of the coagulation factor 9. More than 1,200 mutations have been identified in the F9 gene to date. Point mutations make up approximately 70% of the mutations. In this study, we aimed to determine the F9 gene mutation spectrum in HB patients in Turkey and to contribute to the genotype-phenotype relationships identifying novel mutations. Material and Methods: Fifty five patients who were followed with a diagnosis of HB in 8 different centers and molecularly analyzed in Ege University Faculty of Medicine in November 2018 and January 2020 enrolled to the study. Clinical and laboratory findings of patients were obtained from hospital records. F9 gene sequence analysis was performed using a next generation sequencing platform (MiSeq' Illimuna) Pathogenicity of novel variants were classified according to ACMG 2015. The correlation between mutation distribution and phenotype was evaluated. Results: Among 55 HB patients enrolled in the study, severe HB phenotype were determined in 33 (60%), moderate in 15 (27.3%) and mild in 7 (12.7%). Molecular analysis was revealed 46 different variants in 54 patients (98.2%) of these variants, 30 were missense (63.8%), nine nonsense (19.1%), three frameshift (6.4%), and four splice site (8.5%) mutations. Ten of 46 variants identified has not previously been reported. In one patient no mutation was dertected by sequencing. Conclusion: In this study, the molecular diagnostic success rate and F9 gene mutation spectrum in Turkish HB patients was in accordance with the literature. The results of the study support that HB is a genotypically heterogeneous disease. Ten novel mutations were identified for the first time with this study and contributed to the genotype-phenotype relationship sof the disease.

Keywords: Next generation sequence analysis; hemophilia B; mutation; genotype-phenotype relation
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