Publication: Aterosklerozlu kalp damar hastalığı bulunan bireylerde MMP-1 geni ekzon-1 bölgesindeki tek nükleotit değişikliklerinin sanger dizileme yöntemiyle saptanması
Abstract
Kalp hastalıkları başta olmak üzere beyin ve böbrek damarları gibi dolaşım sistemini etkileyen hastalıklar olarak tanımlanan kardiyovasküler hastalıklar dünyada hastalıklardan kaynaklı ölüm nedenleri dağılımında ilk sıralarda yer almaktadır. Batı toplumlarındahastalıklardan kaynaklı ölüm nedenlerinin yaklaşık %50' sini oluşturan ve damar sertleşmesi olarak adlandırılan ateroskleroz da kardiyovasküler hastalıklarla ilişkili olarak sıklıkla saptanan tablolardan biridir. Günümüze kadar yapılmış bilimsel çalışmalar sonucunda ateroskleroz sürecinin erken dönemlerinde, arteriyel intimada hiperplazi gelişimine ve aterosklerotik lezyonların plak yırtılmasına yol açacak şekilde zayıflaması gibi damarsal bozukluklara sebep olan çeşitli matriks metaloproteinaz türlerinin rolleri olduğu açıklanmıştır. Ancak bu çalışmaların çoğunda Restriksiyon Fragment Uzunluk Polimorfizmi yöntemi adı verilen eski bir yöntem kullanılmakta olup, kullanılan yöntem nükleotit dizilerindeki değişimleri doğrudan gözlemlemek açısından dizi bilgisi sağlamayan biryöntemdir. Ayrıca güncel literatürde matriks metaloproteinaz-1 geninde ekzon-1 tek nükleotit değişikliklerinin ateroskleroz ile ilişkili olup olmadığına dair bir veriye rastlanamamıştır. Bu nedenle çalışmamızda Türkiye popülasyonundaki aterosklerozlubireylerde matriks metaloproteinaz-1 ekzon-1 tek nükleotit değişikliklerinin ateroskleroz tablosu arasındaki korelasyonun saptanabilmesi için aterosklerozlu kalp damar hastalarındaki matriks metaloproteinaz-1 geni ekzon-1 varyantlarının Sanger Dizileme yöntemiyle tespit edilmesi amaçlanmıştır. Çalışmamıza, aterosklerozlu kalp damar hastalığı bulunan 78 bireyden ve herhangi bir kronik rahatsızlığı bulunmayan 5 sağlıklı bireyden alınmış kan örnekleri dahil edilmiştir. Çalışmaya dahil edilen hastalarda aterosklerozlu kalpdamar hastalıkları, kronik obstrüktif akciğer hastalığı, diyabet, hiperkolesterolemi, hipertansiyon, prostat hastalıkları ve paratiroid bezi hastalıkları gibi hastalıklardan biri veya birkaçı bir arada bulunmaktadır. Dizileme çalışmaları sonucunda 83 adet örneğin birindemutasyon şüphesi gözlenmiştir. Mutasyon bulgusuna rastlanan örneğe ait elektroferogram görüntüsünde MMP-1 geni ekzon-1 bölgesinin 106. nükleotidinde G nükleotidinin yanısıra A nükleotidi de gözlenmiştir. Heterezigot olarak saptanan bu durum sinyal peptidininkodlandığı bölgede TGG kodonunun TGA’ya dönüşümüne bağlı olarak bir erken sonlanma kodonunun oluşumuna sebebiyet vermektedir. Bu değişim, NCBI veri tabanına kayıtlı rs1858256060 referans numaralı varyanta karşılık gelmektedir. Ancak MMP-1 polimorfizmlerinin ateroskleroz ve diğer klinik durumlarla ilişkisini tam olarak anlamak için ileri moleküler çalışmalarla birlikte daha fazla klinik incelemenin yapılması gerektiği düşünülmektedir.
Cardiovascular diseases, which are defined as diseases affecting the circulatory system, including heart diseases as well as diseases of the brain and kidney vessels, rank among the leading causes of death from diseases worldwide. In Western societies, atherosclerosis,which is referred to as the hardening of the arteries and accounts for approximately 50% of disease-related deaths, is one of the most frequently observed conditions associated with cardiovascular diseases. Scientific studies conducted to date have demonstrated that various types of matrix metalloproteinases play a role in the development of vascular disorders, such as intimal hyperplasia in the early stages of atherosclerosis and the weakening of atherosclerotic lesions, which can lead to plaque rupture. However, in most of these studies, an old method known as Restriction Fragment Length Polymorphism has been used, which does not provide sequence information necessary for directly observing nucleotide sequence changes. Moreover, no data have been found in the current literature regarding whether single nucleotide variations in exon 1 of the matrix metalloproteinase-1 gene are associatedwith atherosclerosis.Therefore, in this study, we aimed to detect the correlation between single nucleotide variations in exon 1 of the matrix metalloproteinase-1 gene and the presence of atherosclerosis in patients with atherosclerotic cardiovascular disease in the Turkish population using the Sanger sequencing method. Blood samples from 78 individuals with atherosclerotic cardiovascular disease and 5 healthy individuals without any chronic disease were included in our study. In the patients included in the study, one or more of the following conditions are present simultaneously: atherosclerotic cardiovascular diseases, chronic obstructive pulmonary disease, diabetes, hypercholesterolemia, hypertension, diseases of the prostate, and parathyroid gland disorders.Sequencing analyses revealed a suspected mutation in one of the 83 samples. In the electropherogram of the sample where the mutation was detected, it was noted that in addition to the G nucleotide, an A nucleotide was also observed at the 106th nucleotide of the exon-1 region of the MMP-1 gene. Thisheterozygous condition leads to the formation of a premature stop codon due to the conversion of the TGG codon to TGA in the region encoding the signal peptide. This change corresponds to the variant with the reference number rs1858256060 recorded in the NCBI database. However, it is suggested that further molecular studies, as well as additional clinical investigations, are necessary to fully understand the relationship between MMP-1 polymorphisms and atherosclerosis and other clinical conditions.
Cardiovascular diseases, which are defined as diseases affecting the circulatory system, including heart diseases as well as diseases of the brain and kidney vessels, rank among the leading causes of death from diseases worldwide. In Western societies, atherosclerosis,which is referred to as the hardening of the arteries and accounts for approximately 50% of disease-related deaths, is one of the most frequently observed conditions associated with cardiovascular diseases. Scientific studies conducted to date have demonstrated that various types of matrix metalloproteinases play a role in the development of vascular disorders, such as intimal hyperplasia in the early stages of atherosclerosis and the weakening of atherosclerotic lesions, which can lead to plaque rupture. However, in most of these studies, an old method known as Restriction Fragment Length Polymorphism has been used, which does not provide sequence information necessary for directly observing nucleotide sequence changes. Moreover, no data have been found in the current literature regarding whether single nucleotide variations in exon 1 of the matrix metalloproteinase-1 gene are associatedwith atherosclerosis.Therefore, in this study, we aimed to detect the correlation between single nucleotide variations in exon 1 of the matrix metalloproteinase-1 gene and the presence of atherosclerosis in patients with atherosclerotic cardiovascular disease in the Turkish population using the Sanger sequencing method. Blood samples from 78 individuals with atherosclerotic cardiovascular disease and 5 healthy individuals without any chronic disease were included in our study. In the patients included in the study, one or more of the following conditions are present simultaneously: atherosclerotic cardiovascular diseases, chronic obstructive pulmonary disease, diabetes, hypercholesterolemia, hypertension, diseases of the prostate, and parathyroid gland disorders.Sequencing analyses revealed a suspected mutation in one of the 83 samples. In the electropherogram of the sample where the mutation was detected, it was noted that in addition to the G nucleotide, an A nucleotide was also observed at the 106th nucleotide of the exon-1 region of the MMP-1 gene. Thisheterozygous condition leads to the formation of a premature stop codon due to the conversion of the TGG codon to TGA in the region encoding the signal peptide. This change corresponds to the variant with the reference number rs1858256060 recorded in the NCBI database. However, it is suggested that further molecular studies, as well as additional clinical investigations, are necessary to fully understand the relationship between MMP-1 polymorphisms and atherosclerosis and other clinical conditions.