Naci ŞENKAL, Yasemin OYACI, Timurhan CEBECİ, Hilal KONYAOĞLU, Murat KÖSE, Mustafa ÖNEL, Alpay MEDETALİBEYOĞLU, Gözde YEŞİL SAYIN, Mustafa PEHLİVAN, Sacide PEHLİVAN, Ümmuhan İŞOĞLU-ALKAÇ, Tufan TÜKEK

TÜRK POPÜLASYONUNDA COVID-19 DUYARLILIĞI İLE eNOS VARYANTLARININ İLİŞKİSİ

Amaç: Ciddi bir solunum ve damar hastalığı olan COVID-19 hastalığında endotel bariyerinin koruyucu işlevi bozulmaktadır. Nitrik oksid (NO) üretimi için en önemli izoform olan endotelyal NO sentaz (eNOS), çoğunlukla endotel hücrelerinde eksprese edilir. Bu nedenle, bu çalışma, eNOS G894T ve değişken ardışık tekrar sayısı (VNTR) fonksiyonel varyantlarının COVID-19 hastalığının gelişimine yatkınlık oluşturup oluşturmadığını değerlendirmeyi amaçladı. Gereç ve Yöntem: Bu çalışmaya toplam 384 birey (284 COVID-19 hastası ve 100 sağlıklı kontrol) dahil edildi. İki eNOS gen varyantı (G894T ve VNTR), polimeraz zincir reaksiyonu (PCR) ve/ veya kısıtlama fragman uzunluğu polimorfizmi (RFLP) yöntemleri ile genotiplendi. Sonuçlar istatistiksel yöntemlerle değerlendirildi. Bulgular: eNOS G894T ile COVID-19 hastalığı arasında anlamlı bir ilişki tespit edildi. eNOS G894T varyantı için, T/T genotipi ve T aleli taşıyıcılarının COVID-19 hastalığı için artan riske sahip olduğu görüldü (sırasıyla, p=0,035 ve p=0,030), eNOS G894T G/G genotipi, kontrol grubunda hasta grubuna göre daha yaygındı (p=0.030). eNOS VNTR genotipi ve allel frekansları açısından gruplar arasında anlamlı farklılık yoktu (p˃0.05). Bu varyantlar için hasta ve kontrol gruplarının genotipleri HWE’deydi. Sonuç: Bu sonuçlar, eNOS G894T varyantının Türk popülasyonunda artan COVID-19 riski ile ilişkili olduğu hipotezini destekleyen kanıtlar sağlamıştır. Bulgularımız yeni tedavi seçeneklerinin ortaya çıkmasına yol açabilir. Hastalığın patogenezinde yer alan moleküler mekanizmaları anlamak için daha fazla araştırmaya ihtiyaç vardır. Anahtar Kelimeler:

Anahtar Kelimeler:

COVID-19, nitrik oksit sentaz, varyant, RFLP-PCR

ASSOCIATING eNOS GENE VARIANTS WITH COVID-19 SUSCEPTIBILITY IN THE TURKISH POPULATION

Objective: COVID-19 is a serious respiratory and vascular disease that impairs the protective function of the endothelial barrier. Endothelial nitric oxide synthase (eNOS), the most important isoform for nitric oxide (NO) production, is mostly expressed in endothelial cells. Therefore, this study aims to evaluate whether eNOS G894T and variable tandem repeat number (VNTR) functional variants show predisposition to developing COVID-19. Materials and Methods: The study includes a total of 384 subjects (284 COVID-19 patients and 100 healthy controls). Two eNOS gene variants (G894T and VNTR) were genotyped using the polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) methods, with the results being evaluated using statistical methods. Results: A significant association has been identified between eNOS G894T and COVID-19. For the eNOS G894T variant, the T/T genotype (p=0.035) and T allele carriers (p=0.030) appear to have an increased risk of developing COVID-19. The eNOS G894T G/G genotype (p=0.030) was more common in the control group compared to the patient group. No significant difference was found between groups regarding the eNOS VNTR genotype and allele frequencies (p˃0.05). The genotypes of the patient and control groups for these variants were in Hardy-Weinburg equilibrium (HWE). Conclusion: These results provide evidence supporting the hypothesis that the eNOS G894T variant is associated with an increased risk of developing COVID-19 in the Turkish population. These findings may lead to the emergence of new treatment options. Further research is required to understand the molecular mechanisms involved in the pathogenesis of the disease.

Keywords:

COVID-19, nitric oxide synthase, variant, RFLP-PCR,

PDF

___

1. Wiersinga WJ, Rhodes A, Cheng AC, Peacock SJ, Prescott HC. Pathophysiology, transmission, diagnosis, and treatment of coronavirus disease 2019 (COVID-19): a review. JAMA 2020;324(8):782-93 [CrossRef] google scholar
2. Berlin DA, Gulick RM, Martinez FJ. Severe Covid-19. N Engl J Med 2020; 383: 2451-2460. [CrossRef] google scholar
3. Mehta P, McAuley DF, Brown M, Sanchez E, Tattersall RS, Manson JJ and HLH Across Speciality Collaboration UK. COVID-19: consider cytokine storm syndromes and immunosuppression. Lancet 2020;395(10229):1033-4. [CrossRef] google scholar
4. Varga Z, Flammer AJ, Steiger P, Haberecker M, Andermatt R, Zinkernagel AS, et al. Endothelial cell infection and endotheliitis in COVID-19. Lancet 2020;395:1417-8. [CrossRef] google scholar
5. Wu Z, McGoogan JM. Characteristics of and important lessons from the coronavirus disease 2019 (COVID-19) outbreak in China: summary of a report of 72314 cases from the chinese center for disease control and prevention. JAMA 2020;323(13):1239-42. [CrossRef] google scholar
6. Teuwen LA, Geldhof V, Pasut A, Carmeliet P. COVID-19: the vasculature unleashed. Nat Rev Immunol 2020;20(7):389-91. [CrossRef] google scholar
7. Shankarishan P, Borah PK, Ahmed G, Mahanta J. Endothelial nitric oxide synthase gene polymorphisms and the risk of hypertension in an Indian population. Biomed Res Int 2014; 2014:793040. [CrossRef] google scholar
8. Vecoli C. Endothelial nitric oxide synthase gene polymorphisms in cardiovascular disease. Vitam Horm 2014;96:387-406. [CrossRef] google scholar
9. Abdul-Cader MS, Amarasinghe A, Abdul-Careem MF. Activation of toll-like receptor signaling pathways leading to nitric oxide-mediated antiviral responses. Arch Virol 2016;161(8): 2075-86. [CrossRef] google scholar
10. Akerström S, Mousavi-Jazi M, Klingström J, Leijon M, Lundkvist A, Mirazimi A. Nitric oxide inhibits the replication cycle of severe acute respiratory syndrome coronavirus. J Virol 2005;79(3):1966-9. [CrossRef] google scholar
11. Zammiti W, Mtiraoui N, Mahjoub T. Lack of consistent association between endothelial nitric oxide synthase gene polymorphisms, homocysteine levels and recurrent pregnancy loss in Tunisian women. Am J Reprod Immunol 2008;59:139-45. [CrossRef] google scholar
12. Joshaghani HR, Salehi A, Samadian E, Gharei R, Ahmadi AR. Association between NOS3 G894T, T-786C and 4a/4b variants and coronary artery diseases in Iranian population. Iran J Public Health 2018;47(12);1891-8. google scholar
13. Gelzo M, Scialò F, Cacciapuoti S, Pinchera B, De Rosa A, Cernera G, et al. Inducible Nitric Oxide Synthase (iNOS): Why a Different. Viruses 2022;14(3):534. [CrossRef] google scholar
14. Xu S, Ilyas I, Little PJ, Li H, Kamato D, Zheng X, et al. Endothelial dysfunction in atherosclerotic cardiovascular diseases and beyond: from mechanism to pharmacotherapies. Pharmacol Rev 2021;73:924-67. [CrossRef] google scholar
15. Monteil V, Kwon H, Prado P, Hagelkrüys A, Wimmer RA, Stahl M, et al. Inhibition of SARS-CoV-2 infections in engineered human tissues using clinical-grade soluble human ACE2. Cell 2020;181(905-13):e907. [CrossRef] google scholar
16. Fodor A, Tiperciuc B, Login C, Orasan OH, Lazar AL, Buchman C, et al. Endothelial dysfunction, inflammation, and oxidative stress in COVID-19-mechanisms and therapeutic targets. Oxid Med Cell Longev 2021;2021:8671713. [CrossRef] google scholar
17. Coleman JW. Nitric oxide in immunity and inflammation. Int. Immunopharm 2001;1:1397-406. [CrossRef] google scholar
18. Kellner M, Noonepalle S, Lu Q, Srivastava A, Zemskov E, Black SM. ROS signaling in the pathogenesis of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). Adv Exp Med Biol 2017;967:105-37 [CrossRef] google scholar
19. Oliveira-Paula GH Lacchini R, Tanus-Santos JE. Clinical and pharmacogenetic impact of endothelial nitric oxide synthase polymorphisms on cardiovascular diseases. Nitric Oxide 2017;63:39-51. [CrossRef] google scholar
20. Marsden PA, Heng HH, Scherer SW, Stewart RJ, Hall AV, Shi XM, et al. Structure and chromosomal localization of the human constitutive endothelial nitric oxide synthase gene. J Biol Chem 1993;268 (23):17478-88. [CrossRef] google scholar
21. Joshi MS, Mineo C, Shaul PW, Bauer JA. Biochemical consequences of the NOS3 Glu298Asp variation in human endothelium: altered caveolar localization and impaired response to shear. FASEB J 2007;21(11):2655-63. [CrossRef] google scholar
22. Zhang MX, Zhang C, Shen YH, Wang J, Li XN, Zhang Y, et al. Biogenesis of short intronic repeat 27-nucleotide small RNA from endothelial nitricoxide synthase gene. J Biol Chem 2008;283(21):14685-93. [CrossRef] google scholar
23. Oliveira-Paula GH, Lacchini R, Tanus-Santos JE. Endothelial nitric oxide synthase: From biochemistry and gene structure to clinical implications of NOS3 polymorphisms. Gene 2016;575 (2 Pt 3):584-99. [CrossRef] google scholar
24. Zhang MX, Zhang C, Shen YH, Wang J, Li XN, Chen L, et al. Effect of 27 nt small RNA on endothelial nitric-oxide synthase expression. Mol Biol Cell 2008;19:3997-4005. [CrossRef] google scholar
25. Miyamoto Y, Saito Y, Kajiyama N, Yoshimura M, Shimasaki Y, Nakayama M, et al. Endothelial nitric oxide synthase gene is positively associated with essential hypertension. Hypertension 1998;32(1):3-8. [CrossRef] google scholar
26. Diler SB, Öden A. The T -786C, G894T, and Intron 4 VNTR (4a/b) Polymorphisms of the endothelial nitric oxide synthase gene in prostate cancer cases. Genetika 2016;52(2):249-54. [CrossRef] google scholar
27. Yeh CC, Santella RM, Hsieh LL, Sung FC, Tang R. An intron 4 VNTR polymorphism ofthe endothelial nitric oxide synthase gene is associated with early-onset colorectal cancer. Int J Cancer 2009;124(7):1565-71. [CrossRef] google scholar
28. Zhao X, Li Q, Yu F, Lin L, Yin W, Li J, et al. Gene polymorphism associated with endothelial nitric oxide synthase (4VNTR, G894T, C786T) and unexplained recurrent spontaneous abortion risk. Medicine (Baltimore) 2019;98(4):e14175. [CrossRef] google scholar
29. Zhao J, Zhang W, Shen L, Yang X, Liu Y, Gai Z. Association of the ACE, GSTM1, IL-6, NOS3, and CYP1A1 polymorphisms with susceptibility of mycoplasma pneumoniae pneumonia in Chinese children. Medicine (Baltimore) 2017;96(15):e6642. [CrossRef] google scholar
30. Wang QQ, Yu L, Huang GR, Zhang L, Liu YQ, Wang TW, et al. Polymorphisms of angiotensin converting enzyme and nitric oxide synthase 3 genes as risk factors of high-altitude pulmonary edema: a case-control study and meta-analysis. Tohoku J Exp Med 2013;229(4):255-66. [CrossRef] google scholar
31. Li JA, Chen ZB, Lv TG, Han ZL, Liu PP. Impact of endothelial nitric oxide synthase gene polymorphism on severity of enterovirus 71-infection in Chinese children. Clin Biochem 2013;46(18):1842-7. [CrossRef] google scholar
32. Özkan M, Günay N, Sener EF, Karcıoglu Ö, Tahtasakal R, Dal F, et al. Variants in TNF and NOS3 (eNOS) genes associated with sepsis in adult patients. J Gene Med 2021;23 (4):e3323. [CrossRef] google scholar
33. Koskela S, Laine O, Mäkelä S, Pessi T, Tuomisto S, Huhtala H, et al. Endothelial nitric oxide synthase G894T polymorphism associates with disease severity in puumala hantavirus infection. PLoS One 2015;10(11):e0142872. [CrossRef] google scholar
34. Lapic I, Radic Antolic M, Horvat I, Premuzic V, Palic J, Rogic D, et al. Association of polymorphisms in genes encoding prothrombotic and cardiovascular risk factors with disease severity in COVID-19 patients: A pilot study. J Med Virol 2022;94 (8):3669-75. [CrossRef] google scholar
35. Pehlivan S, Köse M, Mese S, Serin I, Senkal N, Oyacı Y, et al. Investigation of MBL2 and NOS3 functional gene variants in suspected COVID-19 PCR (-) patients. Pathog Glob Health 2022;116 (3):178-84. [CrossRef] google scholar
36. Tesauro M, Thompson WC, Rogliani P, Qi L, Chaudhary PP, Moss J. Intracellular processing of endothelial nitric oxide synthase isoforms associated with differences in severity of cardiopulmonary diseases: cleavage of proteins with aspartate vs. glutamate at position 298. Proc Natl Acad Sci U S A 2000;97(6):2832. [CrossRef] google scholar