Mitokondriyal hastalık öntanılı hastalarda tüm ekzom dizi analizi ile moleküler genetik tanının araştırılması ve sorumlu yeni genlerin keşfi
Küçük Resim Yok
Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Giriş: Mitokondriyal hastalıklar (MH), temelde elektron transport zinciri (ETZ) fonksiyon bozukluğuna yol açan hastalıklardır. Sıklığının 1.6/5000 olduğu tahmin edilmektedir ve en sık kalıtımsal nörometabolik hastalık grubudur. Mitkondriyal proteomun 100 civarında proteini direk olarak ETZ ilişkilidir. 300'den fazla nükleer gen defektinin MH'a yol açtığı bilinmektedir. Günümüze kadar edindiğimiz bilgiler ile genotip-fenotip uyumundan sözetmek mümkün değildir. Çalışmamız, mitokondriyal hastalıklarda tanı başarısını arttırmaya yönelik olarak yeni nesil dizi analizi yöntemleri ile fonksiyonel doku analizlerini aynı basamakta birleştirerek tanı başarısını arttırmayı amaçlamaktadır. Yöntem: Ege Üniversitesi Tıp Fakültesi Çocuk Genetik Bilim Dalında mitokondriyal hastalık ön tanısı ile takip edilen, aynı ailede birden fazla etkilenmiş birey olan ve aile ağacı otozomal resesif kalıtım ile uyumlu olan, öncesinde mitokondriyal DNA analizi ve mitokondriyal DNA MLPA analizi ile mitokondriyal DNA ilişkili mitokondriyal hastalıklar dışlanmış olan ancak klinik ve laboratuar bulgular ile (klinik ve laboratuvar) spesifik bir tanı oluşturulamamış olan yaşları 0-18 yaş arasında olan 20 indeks vakaya WES analizi ve eş zamanlı olarak doku örneklemleri alınarak kompleksomik analizi yapılmıştır. Bulgular: Örneklem grubumuzda 10 hastada moleküler ve biyokimyasal olarak tanı konmuştur. Hastalarımızda RRM2B, MTO1, MPV17, NAR2, SDHAF1, COX5A , COQ4, C12Orf65, PDHA1, NDUFAF6 genlerinde toplam 11 varyant saptanmıştır. Saptanan varyantlardan 7/11 varyant ilk kez tanımlanmış, 4/11 varyant daha önce tanımlı varyantlar olarak değerlendirilmiştir. 5/11 varyant missense, 1/11 çerçeve kayması, 2/11 nonsense, 2/11 başlangıç kodonu kaybına neden olan varyantlar olarak saptanmıştır. Kompleksomik analizleri değerlendirildiğinde 7/11 hastada kombine oksidatif fosforilasyon edefekti, 2/11 hastada izole kompleks eksikliği, 2/11 hastada normal olarak saptandı. Bir hastada dokuda kurtarma çalışması yapılarak tanı doğrulaması gerçekleştirildi. Sonuç: Bu çalışmada WES daha önce mtDNA analizi ve konvansiyonel metabolik tarama testleri ile tanı alamamış olan 10 hastanın moleküler etyolojisi belirlendi. MH grubunda yeni nesil dizi analizi yöntemlerinin fonksiyonel analizlerle desteklenmesinin tanı başarısını arttırdığı görülmüştür.
Background: Mitochondrial diseases (MDs) are diseases which are resulted by electron transport chain (ETZ) dysfunction. Its frequency is estimated to be 1.6/5000 and the most common inherited neurometabolic disease group. About 100 proteins of the mitchondrial proteome are directly associated with ETZ. More than 300 nuclear gene defects are known to cause MD. There is no genotype-phenotype correlation with the information we have obtained so far. Our study aims to increase the diagnostic success by combining the next generation sequencing methods and functional tissue analyzes in the same step to increase the diagnostic success in mitochondrial diseases. Method: Mitochondrial DNA-related mitochondrial diseases with mitochondrial DNA analysis and mitochondrial DNA MLPA analysis, which are followed up with a pre diagnosis of mitochondrial disease in Ege University Faculty of Medicine, Department of Pediatrics. WES analysis and tissue samples were taken and complexomic analysis was performed on 20 index cases aged 0-18 years, who were excluded but for whom a specific diagnosis could not be established with clinical and laboratory findings. Results: In our sample group, 10 patients were diagnosed molecularly and biochemically. A total of 11 variants were detected in RRM2B, MTO1, MPV17, NAR2, SDHAF1, COX5A , COQ4, C12Orf65, PDHA1, NDUFAF6 genes in our patients. Among the detected variants, 7/11 variants were defined for the first time, and 4/11 variants were evaluated as previously defined variants. 5/11 variant missense, 1/11 frameshift, 2/11 nonsense, 2/11 variants causing loss of start codon were detected. When complexomic analyzes were evaluated, combined oxidative phosphorylation effect was found in 7/11 patients, isolated complex deficiency was found in 2/11 patients, and normal in 2/11 patients. In one patient, diagnosis was confirmed by performing a tissue study. Conclusion: In this study, the molecular etiology of 10 patients who could not be diagnosed with WES by mtDNA analysis and conventional metabolic screening tests was determined. It has been observed that supporting new generation sequencing methods with functional analyzes increases the success of diagnosis in the MD group.
Background: Mitochondrial diseases (MDs) are diseases which are resulted by electron transport chain (ETZ) dysfunction. Its frequency is estimated to be 1.6/5000 and the most common inherited neurometabolic disease group. About 100 proteins of the mitchondrial proteome are directly associated with ETZ. More than 300 nuclear gene defects are known to cause MD. There is no genotype-phenotype correlation with the information we have obtained so far. Our study aims to increase the diagnostic success by combining the next generation sequencing methods and functional tissue analyzes in the same step to increase the diagnostic success in mitochondrial diseases. Method: Mitochondrial DNA-related mitochondrial diseases with mitochondrial DNA analysis and mitochondrial DNA MLPA analysis, which are followed up with a pre diagnosis of mitochondrial disease in Ege University Faculty of Medicine, Department of Pediatrics. WES analysis and tissue samples were taken and complexomic analysis was performed on 20 index cases aged 0-18 years, who were excluded but for whom a specific diagnosis could not be established with clinical and laboratory findings. Results: In our sample group, 10 patients were diagnosed molecularly and biochemically. A total of 11 variants were detected in RRM2B, MTO1, MPV17, NAR2, SDHAF1, COX5A , COQ4, C12Orf65, PDHA1, NDUFAF6 genes in our patients. Among the detected variants, 7/11 variants were defined for the first time, and 4/11 variants were evaluated as previously defined variants. 5/11 variant missense, 1/11 frameshift, 2/11 nonsense, 2/11 variants causing loss of start codon were detected. When complexomic analyzes were evaluated, combined oxidative phosphorylation effect was found in 7/11 patients, isolated complex deficiency was found in 2/11 patients, and normal in 2/11 patients. In one patient, diagnosis was confirmed by performing a tissue study. Conclusion: In this study, the molecular etiology of 10 patients who could not be diagnosed with WES by mtDNA analysis and conventional metabolic screening tests was determined. It has been observed that supporting new generation sequencing methods with functional analyzes increases the success of diagnosis in the MD group.
Açıklama
Anahtar Kelimeler
Genetik, Genetics
