NLPR3 genin mutasyonlarının veya polimorfizmlerinin genotip − fenotip ilişkisi
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Tarih
2018
Yazarlar
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Yayıncı
Ege Üniversitesi, Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
İnflammasomlar doğal immun yanıtın en önemli bir parçası olup ve adaptif bağışıklığın farklı yönlerini ciddi olarak kontrol eden çoklu hücre içi protein kompleksleridir. Genellikle, inflammasomlar, farklı patojenik (PAMP) veya endojen steril tehlike sinyalleri (DAMP) kabul ettikten sonra ASC gibi adaptör proteinleri kullanarak kaspaz1 aktivasyonu yaparak interlökin (IL) -1β ve IL-18 gibi pro-enflamatuar sitokinlerin işlenmesine ve salınmasına yol açar. Bilinen 7 farkılı inflamazom arasında en çok araştırılan NLRP3 inflamazom yolağıdır. Bu yolağın sensör proteini olan kiryopirini kodlayan NLRP3 geninde saptanan mutasyonların 3 farkılı hastalıgın nedeni olduğu bilinmektedir. Bu çalışmada , otoinflamatuar hastalığa özgü kilinik ve laboratuar bulguları olan , MEFV ve MVK gen mutasyonları negatif olan hastalarda NLRP3 gen analizi yaparak elde edilen DNA varyantlarının ve protein yapısının değerlendirilmesi ve fenotipe yansımasının araştırılması amaçlanmıştır. Bu amaçla 25'i sağlıklı ve 25'i çalışma grubunu oluşturan toplam 50 kişi çalışmaya alınmıştır. Çalışmaya dahil edilen 50 olgudan edle edilen venöz kandan genomik DNA elde edilerek mutasyon analizi sanger DNA dizileme yöntemleri ile ve sonuçların okunması ise Finch TV, CLC Sequence Viewer - QIAGEN Bioinformatics ve MEGA7 biyanformatik porogramları ile yapılmıştır. Mutsyon değerlendirmeleri NCBI, PolyPhen-2, İNFEVERS ve bir çok farkılı veritabanlarına göre yapılmiştir. Kontrol ve çalışma gurubunda iki adet (Ala240Ala, Arg260Arg ) sinonim amino asit mutasyonu saptanmıştır ve fenotip ile hiç bir ilişkisi olmadiği için polimorfizm olarak değerlendirilmiştir. Ayrıca çalışma gurubunda iki kişide sekans taranmasında mutasyon bulunmadı ve mühtemenel diğer genetik Otoinflamatuvar hastalık açısından diğer genlerin çalışılması yapıldı. Kontrol gurubunda hastalıkla ilişkili hiç bir mutasyon saptanmamıştır. Çalışma gurubunda hastalıkla iklişkili 6 adet farklı mutasyon saptanmıştır bunlardan biri Thr913Met mutasyon olmak üzere tarafımızdan ilk defa olarak İNFEVERS veritabanına bildirilmiştir. Çalışma grubunda ayrıca 2 adet sinonim aminoasit mutasyonu da saptanmıştır. Her mutasyonun protein yapısına etkisi ayrıca araştırılmıştır .Sonuç olarak, bu tezde , hem sağlıklı hem de otoinflamatuar kilinik ve labratuar bulguları olan hastalarda NLRP3 gen analizi yapılarak hastalığın nedensel genetik varyantıları saptanmış ve proteine etkileri ortaya konulmuştur.
Inflammomas are an essential part of the natural immune response and are multiple intracellular protein complexes that severely control different aspects of adaptive immunity. Generally, sensory molecules recognize different pathogenic (PAMP) or endogenous sterile distress signals (DAMP) they activate caspase-1 using adapter proteins such as ASC, leading to the processing and release of pro-inflammatory cytokines such as interleukin (IL) -1β and IL-18. 7 different, the inflammasome is known among my most researched NLRP3 inflammasome pathway. It is known that mutations detected in the NLRP3 gene, which encodes the sensor protein chryphoria, are caused by 3 different diseases. In this study, it was aimed to evaluate the DNA variants and protein structure obtained by NLRP3 gene analysis in patients with clinically and laboratory findings specific for autoinflammatory disease and negative MEFV and MVK gene mutations, and to investigate its reflection on the phenotype. For this purpose, a total of 50 people, 25 of which are healthy and 25 of which are working groups, were taken into employment. Mutagenesis was performed by sanger DNA sequencing methods and results were read by Finch TV, CLC Sequence Viewer - QIAGEN Bioinformatics and MEGA7 biomorphological porograms. Mortality evaluations were done according to NCBI, PolyPhen-2, INFEVERS and many different databases. Consequently, control and two study groups (Ala240Ala and Arg260Arg) synonymous amino acid mutations were identified and evaluated with no phenotype poyimorfizm as to whether there is a relationship. In addition, two individuals in the study group did not have a mutation in the Intron-Exon sequence screening and it is estimated that other genes associated with the genetic genetic autoinflammatory disease were affected. There was no disease-related mutation in the control group. Seven different mutations were identified in the study group, which were reported to the INFEVERS database for the first time as a Thr913Met mutation. The effect of each mutation on protein structure was further investigated. Consequently, in this thesis, NLRP3 gene analysis was performed in patients with both healthy and autoinflamatory clinical and laboratory findings and the causative genetic variants of the disease were determined and protein effects were determined.
Inflammomas are an essential part of the natural immune response and are multiple intracellular protein complexes that severely control different aspects of adaptive immunity. Generally, sensory molecules recognize different pathogenic (PAMP) or endogenous sterile distress signals (DAMP) they activate caspase-1 using adapter proteins such as ASC, leading to the processing and release of pro-inflammatory cytokines such as interleukin (IL) -1β and IL-18. 7 different, the inflammasome is known among my most researched NLRP3 inflammasome pathway. It is known that mutations detected in the NLRP3 gene, which encodes the sensor protein chryphoria, are caused by 3 different diseases. In this study, it was aimed to evaluate the DNA variants and protein structure obtained by NLRP3 gene analysis in patients with clinically and laboratory findings specific for autoinflammatory disease and negative MEFV and MVK gene mutations, and to investigate its reflection on the phenotype. For this purpose, a total of 50 people, 25 of which are healthy and 25 of which are working groups, were taken into employment. Mutagenesis was performed by sanger DNA sequencing methods and results were read by Finch TV, CLC Sequence Viewer - QIAGEN Bioinformatics and MEGA7 biomorphological porograms. Mortality evaluations were done according to NCBI, PolyPhen-2, INFEVERS and many different databases. Consequently, control and two study groups (Ala240Ala and Arg260Arg) synonymous amino acid mutations were identified and evaluated with no phenotype poyimorfizm as to whether there is a relationship. In addition, two individuals in the study group did not have a mutation in the Intron-Exon sequence screening and it is estimated that other genes associated with the genetic genetic autoinflammatory disease were affected. There was no disease-related mutation in the control group. Seven different mutations were identified in the study group, which were reported to the INFEVERS database for the first time as a Thr913Met mutation. The effect of each mutation on protein structure was further investigated. Consequently, in this thesis, NLRP3 gene analysis was performed in patients with both healthy and autoinflamatory clinical and laboratory findings and the causative genetic variants of the disease were determined and protein effects were determined.
Açıklama
Anahtar Kelimeler
NLRP3 Geni, CAPS Hastalığı, İnflamazom, Mutasyon, Sanger Dizileme, Polimorfizim, Fenotip, NLRP3, GENE, CAPS, Genetic, Auto-inflammatory
