Potential Contribution to Antidiabetic Treatments of Epstein-Barr Virus Proteins LMP1 and EBNA1

Abstract

The prevalence and incidence of diabetes is a global healthcare-management problem that is increasing almost every day. The high numbers of complications make the diagnosis and treatment of diabetes extremely important. Nowadays, it is believed that a large number of diseases, including diabetes, are related to the Endoplasmic Reticulum (ER) stress-induced apoptosis. Proliferative activity of Epstein-Barr Virus (EBV) proteins occurs via two mediators Epstein-Barr Nuclear Antigen-1 (EBNA1) and Latent Membrane Protein-1 (LMP1). In this study, it was aimed to investigate the reversal of pancreatic cell loss by applying recombinant viral EBV proteins. It has also been investigated how related proteins can effect DNA damage, ER stress, and contribute to insulin synthesis-release. For these targets, it has been decided to use doses of 0.1 ppm and 0.5 ppm of commercially obtained recombinant EBNA1 and LMP1 proteins. The possible effects of these proteins on mRNA expression levels of genes known to be responsible for the specified proliferation have been investigated. In the study, DNA damage was analyzed by Comet Assay, and Endoplasmic Reticulum stress was analyzed by Real Time RT-PCR. In RT-PCR studies, mRNA expression levels of cell proliferation markers including PDX 1, WNT 4, FoxO1, TCF7L2 and beta catenin genes in the EBNA1 group were increased by 6.48, 4.25, 2.37, 3.8 and 1.4 times, respectively compared with control group. At the end of the study; EBV recombinant proteins LMP1 and EBNA1 have been shown to increase the expression levels of genes responsible for proliferation. It has also been shown that related proteins may be advantageous in reversing beta cell losses by suppressing the expression levels of ER stress-inducing genes.  It has been thought that the use of viro-therapeutic agents in antidiabetic treatments may be appropriate if supported by higher-level studies in this regard.

Keywords

• Epstein Barr Virus,EBNA1,LMP1,Diabetes

Epstein-Barr Virus Proteinleri LMP1 ve EBNA1’in Antidiyabetik Tedavilere Potensiyel Katkısı

Abstract

Diyabet prevalansı ve insidansı her geçen gün artmaya devam eden global bir sağlık problemidir. Diyabetik komplikasyonların sıklığı diyabet tanı ve tedavisini önemli kılmaktadır. Günümüzde, diyabet de dahil olmak üzere çok sayıda hastalığın Endoplazmik Retikülüm (ER) stres kaynaklı apoptoz ile ilişkili olduğuna inanılmaktadır. Epstein-Barr virusun proliferatif etkinliği; Epstein-Barr Nuclear Antigen-1 (EBNA1) ve Latent Membrane Protein-1 (LMP1) adlı iki mediatör protein aracılığıyla olmaktadır.  Bu çalışmada; pankreatik beta hücre kayıplarının bertarafında  iligili EBV proteinlerinin kullanılması amaçlanmıştır. Ayrıca ilgili proteinlerin ER stresi, insulin sentez ve salınımı ve DNA hasarı üzerine etkilerinin araştırılması hedeflenmiştir. Bu hedefler için, 0,1 ppm ve 0,5 ppm dozlarında ticari olarak elde edilen rekombinant EBNA1 ve LMP1 proteinleri kullanılmıştır. Bu proteinlerin proliferasyondan sorumlu olduğu bilinen genlerin mRNA ekspresyon seviyeleri üzerindeki olası etkileri araştırılmıştır. Çalışmada DNA hasarı Comet Assay ile, proliferasyon düzeyleri Real Time RT-PCR ile analiz edilmiştir. RT-PCR çalışmasında, hücre proliferasyonundan sorumlu oldukları bilinen PDX 1,WNT 4, FoxO1, TCF7L2 ve  beta katenin genlerinin mRNA ekpresyon düzeyleri control grubana nazaran EBNA 1 grubunda sırasıyla 6.48, 4.25, 2.37, 3.8 ve 1.4 kat arttığı tespit edilmiştir. Aynı zamanda ilgili proteinlerin ER stres kaynaklı genlerin ekspresyon seviyelerini bastırarak beta hücre kayıplarını tersine çevirmek açısından avantajlı olabileceği gösterilmiştir. Bu proteinlerin antidiyabetik tedavilerde viro-terapötik ajan olarak kullanılması bu konuda daha üst düzey çalışmalarla desteklenmeye muhtaçtır.

Keywords

• Epstein Barr Virus,EBNA1,LMP1,Diyabet

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