Respiratuvar distres sendromlu preterm yenidoğanlarda NKX2.1 geninin yeni nesil dizi analizi ile araştırılması
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Tarih
2022
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Düzce Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Giriş-Amaç: Respiratuvar distres sendromu (RDS), tedavisinde ciddi gelişmeler elde edilmesine rağmen, erken doğan bebeklerde solunum sıkıntısının en sık rastlanan nedenidir ve preterm doğum ile ilişkili en önemli mortalite ve morbidite sebebidir. RDS insidansı 28 hafta ve daha küçük prematürlerde %93 olup, görülme sıklığı bebeğin gebelik yaşı ile ters orantılı olarak artmaktadır. Patofizyolojisinde prematüre bebeklerdeki yetersiz pulmoner sürfaktan üretiminin neden olduğu gösterildikten sonra, 1959 yılında hastalığın eski adı olan "hiyalen membran hastalığı" RDS ile değiştirildi. Son zamanlarda yapılan epidemiyolojik çalışmalarda; ailevi eğilimin ve ırksal farklılıkların RDS riskini arttırdığının görülmesi üzerine genetik araştırmalara eğilim artmıştır. Aynı gebelik haftasında doğan tüm bebekler için; antenatal steroid uygulamasına, postnatal sürfaktan tedavisine ve optimal ventilatör bakımına rağmen tedaviye eşit yanıtın alınmadığı görülmüştür. Pulmoner sürfaktan proteinlerinin homeostazında rol oynayan NKX2.1 (NK2 homeobox 1) geni ayrıca akciğer farklılaşmasının erken bir belirteci olarak ve akciğerin yapısal gelişimi ile yüzey aktif madde proteinleri SP-B, SP-C ve ABCA3'ün ekspresyonu için önemlidir. NKX2.1'deki mutasyonlar, yenidoğan bebeklerde RDS ve büyük çocuklarda interstisyel akciğer hastalığı, ve solunum yetmezliğine neden olur. Çalışmamızda RDS 'li preterm yenidoğanlarda NKX2.1 geninde yeni nesil dizi analizi yöntemiyle etiyolojiden sorumlu genetik değişimi ortaya koymayı hedefledik. Böylece hastalığın oluşumundan sorumlu tutulan genlerden olan NKX2.1 geninde meydana gelen mutasyonlar göz önünde bulundurularak uygulanılacak olan terapötik yaklaşımlar ile tedavi maliyetinin azaltılmasının yanı sıra hastalar için yan etkilerin daha az olduğu yeni tedavi stratejilerinin geliştirilmesine katkı sağlamayı amaçladık. Gereç-Yöntem: Çalışmaya Düzce Üniversitesi Tıp Fakültesi Eğitim ve Araştırma Hastanesi, Çocuk Sağlığı ve Hastalıkları Anabilim Dalı Yenidoğan Yoğun Bakım Ünitesi'nde prematüre doğum ve RDS nedeni ile izlenen 32 hasta ve RDS dışı başka nedenlerle takip edilen 18 prematüre yenidoğan kontrol grubu olarak alındı. Olguların hedeflenen moleküler genetik tanısı için ilk olarak rutin amaçlı istenilen tam kan örneklerinden geriye kalan atık kanlardan DNA izolasyon işlemi yapıldı. İzole edilen DNA örneklerinden hedeflenen gen bölgeleri, spesifik primerler kullanılarak çoğaltılıp saflaştırma işlemi uygulandıktan sonra hedefe yönelik yeni nesil dizi analizi yöntemiyle bu mutasyonlar incelendi. Çalışmada yer alan preterm yenidoğanlarda c.-61C>T heterozigot, c.-85G>T heterozigot, c.446A>C heterozigot, c.1031G>A p.Gly344Asp heterozigot varyasyonları ve NKX2.1 geninde bileşik heterozigot mutasyonları saptandı. Bulgular: RDS tanılı 32 hastanın 20'si (%62,5), erkek 12'si (%37,5) kız olup erkek/kız oranı 1,66 idi. Kontrol grubundaki 18 hastanın 8'i (%44,4) erkek, 10'u (%55,6) kız olup erkek/kız oranı 0,8 olarak bulundu. Hasta grubunun gestasyonel yaşı (hafta) ortalama 32,71±2.03 hafta, Kontrol grubunun 33,05±7.05 hafta olduğu saptandı. Antropometrik değerler incelendiğinde iki grup arasında doğum ağırlığı, boy ve baş çevresi değerleri açısından anlamlı bir farklılık yoktu. Hastanede yatış süresi ve entübe takip süresi Hasta grubunda istatistiksel olarak anlamlı bir şekilde Kontrol grubundan fazlaydı. Anneye ait parametreler incelendiğinde anne yaşı, gravite, parite, abortus sayısı ve antenatal steroid uygulanması açısından Hasta ve Kontrol Grubunda anlamlı bir fark tespit edilmedi. Çalışmaya dahil edilen preterm yenidoğanlarda c.-61C>T heterozigot, c.-85G>T heterozigot, c.446A>C heterozigot, c.1031G>A p.Gly344Asp heterozigot varyasyonları ve NKX2.1 geninde bileşik heterozigot mutasyonu saptandı. Sonuç: NKX2.1 genindeki c.1031G>A p.Gly344Asp varyasyonu literatürde RDS'li hastalarda daha önce hiç tanımlanmamış olup, bu Çalışmamızda ilk defa tespit edilmiş olması ve bileşik heterozigot mutasyonu gibi yalnızca RDS'li gruptaki yenidoğanlarda rastlanılmış olmaları nedeniyle tespit edilen bu değişikliklerin RDS için belirteç olarak kullanılabileceğini düşünüyoruz. Anahtar Kelimeler: Respiratuar distres sendromu, prematüre, yeni nesil dizi analizi, NK2 homeobox 1 geni
IV.ABSTRACT Screening of the NKX2.1 gene in Preterm Newborns with Respiratory Distress Syndrome by Next Generation Sequencing Introduction-Aim: Respiratory distress syndrome (RDS) is the most common cause of respiratory distress in preterm infants and the most important cause of mortality and morbidity associated with preterm birth, despite significant advances in its treatment. The incidence of RDS is 93% in preterms at gestastional age of 28 weeks and younger, and its incidence increases inversely with the gestational age of the baby. After its pathophysiology was shown to be caused by insufficient pulmonary surfactant production in premature infants, the old name of the disease "hyaline membrane disease" was replaced by RDS in 1959. In recent epidemiological studies; While familial and racial differences were seen to increase the risk of RDS, the tendency to genetic research has increased. For all babies born in the same gestational week; it was observed that there was no equal response to treatment, despite antenatal steroid administration, postnatal surfactant treatment and optimal ventilator care,. The NKX2.1 (NK2 homeobox 1) gene, which plays a role in homeostasis of pulmonary surfactant proteins, is also important as an early marker of lung differentiation and structural development of the lung and expression of surfactant proteins SP-B, SP-C and ABCA3. Mutations in NKX2.1 gene cause RDS in newborn infants and interstitial lung disease and respiratory failure in older children. In our study, we aimed to reveal the genetic change responsible for the etiology in the NKX2.1 gene in preterm newborns with RDS by next generation sequencing method. Thus, we aimed to contribute to the development of new treatment strategies with less side effects for patients, as well as reducing the cost of treatment with the therapeutic approaches to be applied by considering the mutations in the NKX2.1 gene, which is one of the genes responsible for the formation of the disease. Material-Method: 32 patients of premature newborns with RDS followed up in Düzce University Medical Faculty Training and Research Hospital, Department Pediatrics, Neonatal Intensive Care Unit, and 18 premature newborns followed up for other reasons of RDS were included in the study. In order to perform the targeted molecular genetic diagnosis of the cases, first of all, DNA isolations were performed from the remaining waste blood from the whole blood samples requested for routine purposes. After the targeted gene regions from the isolated DNA samples were amplified by using specific primers and purified, these mutations were examined by the next generation sequencing method. C.-61C>T heterozygous, c.-85G>T heterozygous, c.446A>C heterozygous, c.1031G>A p.Gly344Asp heterozygous variations and compound heterozygous mutations in the NKX2.1 gene were found in the preterm newborns included in the Study groups. Results: Twenty (62.5%) of 32 patients with RDS were male and 12 (37.5%) were female. The male/female ratio was calculated as 1.66. While the mean week of gestastional age at delivery in the Patient group was 32,719±2.03 weeks, it was calculated as 33,056±7.058 weeks in the Control group. When the anthropometric values were examined, there was no significant differences between the two groups in terms of birth weight, height and head circumference. Length of hospital stay and intubated follow-up time were statististical significantly higher in the Patient group than the Control group. When the maternal parameters were examined, no significant differences were found in the Patient and Control groups in terms of maternal age, gravity, parity abortion number and antenatal steroid administration. c.-61C>T heterozygous, c.-85G>T heterozygous, c.446A>C heterozygous, c.1031G>A p.Gly344Asp heterozygous variations and compound heterozygous mutation in the NKX2.1 gene were detected in the newborns included in the Study group. Conclusion: The c.1031G>A p.Gly344Asp variations in the NKX2.1 gene have never been described in the literature in patients with RDS, and these changes, such as compound heterozygous mutation, were detected only in newborns in the RDS group, we found thatthis mutations were detected for the first time in our study. We think it may be used as a marker for RSD. Keywords: Respiratory distress sendrome, premature, next generation sequencing analysis, NK2 homeobox 1 gene
IV.ABSTRACT Screening of the NKX2.1 gene in Preterm Newborns with Respiratory Distress Syndrome by Next Generation Sequencing Introduction-Aim: Respiratory distress syndrome (RDS) is the most common cause of respiratory distress in preterm infants and the most important cause of mortality and morbidity associated with preterm birth, despite significant advances in its treatment. The incidence of RDS is 93% in preterms at gestastional age of 28 weeks and younger, and its incidence increases inversely with the gestational age of the baby. After its pathophysiology was shown to be caused by insufficient pulmonary surfactant production in premature infants, the old name of the disease "hyaline membrane disease" was replaced by RDS in 1959. In recent epidemiological studies; While familial and racial differences were seen to increase the risk of RDS, the tendency to genetic research has increased. For all babies born in the same gestational week; it was observed that there was no equal response to treatment, despite antenatal steroid administration, postnatal surfactant treatment and optimal ventilator care,. The NKX2.1 (NK2 homeobox 1) gene, which plays a role in homeostasis of pulmonary surfactant proteins, is also important as an early marker of lung differentiation and structural development of the lung and expression of surfactant proteins SP-B, SP-C and ABCA3. Mutations in NKX2.1 gene cause RDS in newborn infants and interstitial lung disease and respiratory failure in older children. In our study, we aimed to reveal the genetic change responsible for the etiology in the NKX2.1 gene in preterm newborns with RDS by next generation sequencing method. Thus, we aimed to contribute to the development of new treatment strategies with less side effects for patients, as well as reducing the cost of treatment with the therapeutic approaches to be applied by considering the mutations in the NKX2.1 gene, which is one of the genes responsible for the formation of the disease. Material-Method: 32 patients of premature newborns with RDS followed up in Düzce University Medical Faculty Training and Research Hospital, Department Pediatrics, Neonatal Intensive Care Unit, and 18 premature newborns followed up for other reasons of RDS were included in the study. In order to perform the targeted molecular genetic diagnosis of the cases, first of all, DNA isolations were performed from the remaining waste blood from the whole blood samples requested for routine purposes. After the targeted gene regions from the isolated DNA samples were amplified by using specific primers and purified, these mutations were examined by the next generation sequencing method. C.-61C>T heterozygous, c.-85G>T heterozygous, c.446A>C heterozygous, c.1031G>A p.Gly344Asp heterozygous variations and compound heterozygous mutations in the NKX2.1 gene were found in the preterm newborns included in the Study groups. Results: Twenty (62.5%) of 32 patients with RDS were male and 12 (37.5%) were female. The male/female ratio was calculated as 1.66. While the mean week of gestastional age at delivery in the Patient group was 32,719±2.03 weeks, it was calculated as 33,056±7.058 weeks in the Control group. When the anthropometric values were examined, there was no significant differences between the two groups in terms of birth weight, height and head circumference. Length of hospital stay and intubated follow-up time were statististical significantly higher in the Patient group than the Control group. When the maternal parameters were examined, no significant differences were found in the Patient and Control groups in terms of maternal age, gravity, parity abortion number and antenatal steroid administration. c.-61C>T heterozygous, c.-85G>T heterozygous, c.446A>C heterozygous, c.1031G>A p.Gly344Asp heterozygous variations and compound heterozygous mutation in the NKX2.1 gene were detected in the newborns included in the Study group. Conclusion: The c.1031G>A p.Gly344Asp variations in the NKX2.1 gene have never been described in the literature in patients with RDS, and these changes, such as compound heterozygous mutation, were detected only in newborns in the RDS group, we found thatthis mutations were detected for the first time in our study. We think it may be used as a marker for RSD. Keywords: Respiratory distress sendrome, premature, next generation sequencing analysis, NK2 homeobox 1 gene
Açıklama
Anahtar Kelimeler
Respiratuar distres sendromu, prematüre, yeni nesil dizi analizi, NK2 homeobox 1 geni, Respiratory distress sendrome, premature, next generation sequencing analysis, NK2 homeobox 1 gene, Çocuk Sağlığı ve Hastalıkları, Child Health and Diseases, Pulmoner sürfaktanlar, Pulmonary surfactants
