Astımlı hastalarda interlökin-4 (IL4) ve lökotrien C4 sentaz (LTC4s) genlerinin yeni nesil dizi analizi ile araştırılması
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Dosyalar
Tarih
2020
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Düzce Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Giriş: Astım farklı uyaranlara kars?ı artmıs? havayolu duyarlılığı ve geri dönüs?ümlü havayolu obstrüksiyonu ile karakterize, genetik ve çevresel risk faktörlerinin birles?iminden kaynaklanan tekrarlayan kronik inflamatuar bir hastalıktır. Çocukluk çağı kronik hastalıklarının en sık görülenidir. Günümüze kadar sürekli olarak prevalansı, morbiditesi ve mortalitesi artan bir hastalık haline gelmis?tir. Astım olus?umunda üç aşama sorumlu tutulmaktadır. Bunlar, artmıs? havayolu duyarlılığı, havayolu inflamasyonu ve geri dönüs?ümlü havayolu obstrüksiyonudur. Astım multifaktöryel bir hastalık olarak değerlendirilmektedir. Astımlı ebeveynlerin çocuklarında astım riski arttığı uzun zamandır bilinmektedir ve astım prediktivite indeksinin üç majör risk faktöründen biridir. Astım, basit Mendelian kalıtım sergilemeyen karmas?ık genetik yapıya sahip bir hastalıktır. Astım genetiği ile ilgili günümüzde yapılan birçok çalıs?ma mevcuttur, ama hala net olarak astım genetiği anlas?ılamamıs?tır. Bildiğimiz kadarıyla literatürde IL-4 veya LTC4S geninin yeni nesil dizi analizi yöntemi ile değerlendirildiği çalışma yoktur. Biz bu çalışma ile astım tanısıyla takip edilen hastalarda yeni nesil dizi analizi ile IL-4 ve LTC4 genindeki değişimleri ortaya koymayı amaçladık. Gereç ve Yöntem: Bu çalışma Düzce Üniversitesi Tıp Fakültesi Çocuk Sağlığı ve Hastalıkları Kliniğinde GINA kriterlerine göre astım tanısı konulup takip edilen 50 hasta üzerinde yapılmış prospektif bir çalışmadır. İlk olarak rutin amaçlı istenilen tam kan örneklerinden geriye kalan atık kanlardan DNA izolasyon işlemi yapıldı. İzole edilen DNA örneklerinden hedeflenen gen bölgeleri, bu bölgelere spesifik primerler kullanılarak çoğaltılıp saflaştırma işlemi uygulandıktan sonra hedefe yönelik yeni nesil dizi analizi yöntemiyle bu bölgelerdeki mutasyonlar saptandı. Ayrıca hastaların demografik, klinik ve laboratuvar verileri kaydedildi. Bulgular: Çalıs?maya 32'si (%64) erkek, 18'si (%36) kız olmak üzere toplam 50 hasta alındı. Hastaların yas?ı ortalama 7,12±3,8 yıl (1-16 yıl), semptom bas?lama yas?ı ise ortalama 2,73±3,2 yıl (3 ay-13 yıl) idi. Çalışmamıza dahil edilen 50 hastanın 31'inde (%62) mutasyon saptanmıştır. Astımlı olgu grubunda IL-4 geninde intron 1'de c.-33C>T, intron 3'de c.361-9C>A, ekzon 4'te c.23G>A ve intron 3'de c.360+18 C>A mutasyonları saptandı. LTC4S geninde intron 4'te c.312-16 T>C, intron 3'de c.230-12 T>C, intron 1'de c.59-10 C>A, intron 1'de c.-33 C>T ve intron 3'de c.361-9 C>A mutasyonları tanımlandı. Sonuçlar: Astıma en sık es?lik eden hastalık allerjik rinit ve alerjik konjonktivit olarak değerlendirildi. Astım tanısı koyarken MPV düs?üklüğünün kriterlerden biri olabileceği kanaatindeyiz. Total IgE düzeylerinin astım tanısında önemli parametrelerden birisi olduğu çalıs?mamızda da görüldü. IL-4 geninde mutasyon tas?ıyıcı bireylerde akut ürtiker olus?ma olasılığı, tas?ıyıcı olmayanlara göre 5,6 kat daha fazla olduğu görüldü. IL-4 geninde homozigot olarak mutasyon saptanan olguların kliniğinin heterozigot olanlara göre daha kötü olması IL-4 gen mutasyonlarının astım ile ilis?kili olduğunu göstermis?tir. Çalıs?mamızda LTC4S geninde yüksek oranda gözlenen intron 4'te c.312-16 T>C genetik değis?imi astımla ilis?kilendirildi. Çalışmamızın astımın genetik alt yapısı ve farmakogenetik konusu üzerine yapılacak daha geniş ve kontrollü çalışmalara ışık tutacağını umuyoruz.
Introduction: Asthma is a recurrent chronic inflammatory disease caused by a combination of genetic and environmental risk factors, characterized by increased airway sensitivity to different stimuli and reversible airway obstruction. It is the most common childhood chronic respiratory disease. To date, its prevalence, morbidity and mortality have become increasingly common. Three stages are held responsible for the occurrence of asthma. These are increased airway sensitivity, airway inflammation and reversible airway obstruction. Asthma is considered as a multifactorial disease. The risk of asthma in children of asthmatic parents has long been known and is one of the three major risk factors of the asthma predictive index. Asthma is a complex genetic disease that does not exhibit simple Mendelian inheritance. There are many studies on asthma genetics, but asthma genetics is still not clearly understood. To the best of our knowledge, there are no studies in the literature in which the IL-4 or LTC4S gene have been investigated by the next generation sequence analysis method. In this study, we aimed to reveal the changes in IL-4 and LTC4 genes by the next generation sequencing in patients followed up with asthma diagnosis. Materials and Method: This study is a prospective study conducted on 50 patients who were diagnosed with asthma according to GINA criteria and followed at the Pediatrics Clinic of Düzce University Medical Faculty. DNA isolation was performed from the remaining blood after routine whole blood samples. The targeted gene regions from the isolated DNA samples were amplified and purified using primers specific to these regions and mutations in these regions were determined by targeted next generation sequencing. In addition, demographic, clinical and laboratory data of the patients were recorded. Results: A total of 50 patients, 32 (64%) male and 18 (36%) female, were included in this study. The mean age of the patients was 7.12±3.8 years (1-16 years) and the mean age of onset of symptoms was 2.73±3.2 years (3 months-13 years). Mutations were detected in 31 (62%) of the 50 patients included in our study. Mutations detected in the IL-4 gene were c.-33C>T in intron 1, c.3619 C>A in intron 3, c.23 G>A in exon 4, and c.360+18 C>A in intron 3. Mutations identified in the LTC4S gene were c.312-16 T>C in intron 4, c.230-12 T>C in intron 3, c.59-10 C>A in intron 1, c.-33 C>T in intron 1, and c.361-9 C>A in intron 3. Discussion: Allergic rhinitis and allergic conjunctivitis were the most common comorbidities of asthma. We believed that low MPV might be one of the criteria in the diagnosis of asthma. Total IgE levels were found to be one of the important parameters in the diagnosis of asthma. The probability of acute urticaria in mutation carriers of the IL-4 gene was 5.6 times higher than in non-carriers. The fact that cases with homozygous mutations in the IL-4 gene were worse than those with heterozygous Clinical findings in asthmatic cases showed that IL-4 gene mutations were associated with asthma. In our study, c.312-16 T> C genetic change in intron 4, which is highly observed in LTC4S gene, was associated with asthma. We hope that our study will shed light on larger and controlled studies on the genetic background and pharmacogenetics of asthma.
Introduction: Asthma is a recurrent chronic inflammatory disease caused by a combination of genetic and environmental risk factors, characterized by increased airway sensitivity to different stimuli and reversible airway obstruction. It is the most common childhood chronic respiratory disease. To date, its prevalence, morbidity and mortality have become increasingly common. Three stages are held responsible for the occurrence of asthma. These are increased airway sensitivity, airway inflammation and reversible airway obstruction. Asthma is considered as a multifactorial disease. The risk of asthma in children of asthmatic parents has long been known and is one of the three major risk factors of the asthma predictive index. Asthma is a complex genetic disease that does not exhibit simple Mendelian inheritance. There are many studies on asthma genetics, but asthma genetics is still not clearly understood. To the best of our knowledge, there are no studies in the literature in which the IL-4 or LTC4S gene have been investigated by the next generation sequence analysis method. In this study, we aimed to reveal the changes in IL-4 and LTC4 genes by the next generation sequencing in patients followed up with asthma diagnosis. Materials and Method: This study is a prospective study conducted on 50 patients who were diagnosed with asthma according to GINA criteria and followed at the Pediatrics Clinic of Düzce University Medical Faculty. DNA isolation was performed from the remaining blood after routine whole blood samples. The targeted gene regions from the isolated DNA samples were amplified and purified using primers specific to these regions and mutations in these regions were determined by targeted next generation sequencing. In addition, demographic, clinical and laboratory data of the patients were recorded. Results: A total of 50 patients, 32 (64%) male and 18 (36%) female, were included in this study. The mean age of the patients was 7.12±3.8 years (1-16 years) and the mean age of onset of symptoms was 2.73±3.2 years (3 months-13 years). Mutations were detected in 31 (62%) of the 50 patients included in our study. Mutations detected in the IL-4 gene were c.-33C>T in intron 1, c.3619 C>A in intron 3, c.23 G>A in exon 4, and c.360+18 C>A in intron 3. Mutations identified in the LTC4S gene were c.312-16 T>C in intron 4, c.230-12 T>C in intron 3, c.59-10 C>A in intron 1, c.-33 C>T in intron 1, and c.361-9 C>A in intron 3. Discussion: Allergic rhinitis and allergic conjunctivitis were the most common comorbidities of asthma. We believed that low MPV might be one of the criteria in the diagnosis of asthma. Total IgE levels were found to be one of the important parameters in the diagnosis of asthma. The probability of acute urticaria in mutation carriers of the IL-4 gene was 5.6 times higher than in non-carriers. The fact that cases with homozygous mutations in the IL-4 gene were worse than those with heterozygous Clinical findings in asthmatic cases showed that IL-4 gene mutations were associated with asthma. In our study, c.312-16 T> C genetic change in intron 4, which is highly observed in LTC4S gene, was associated with asthma. We hope that our study will shed light on larger and controlled studies on the genetic background and pharmacogenetics of asthma.
Açıklama
YÖK Tez No: 631797
Anahtar Kelimeler
Çocuk Sağlığı ve Hastalıkları, Child Health and Diseases
