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Endothelial Nitric Oxide Synthase (eNOS) Gene Polymorphisms and Markers of Hemolysis, Inflammation and Endothelial Dysfunction in Brazilian Sickle Cell Anemia Patients

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Abstract

The impaired bioavailability of endogenous nitric oxide (NO) in sickle cell anemia (SCA) may be influenced by polymorphisms in the endothelial nitric oxide synthase gene (eNOS). We compared allelic/genotypic frequencies of the eNOS polymorphisms T-786C, VNTR4a/b and G894T between 89 adult SCA patients and 100 healthy controls, and investigated the relationship between these SNPs and markers of hemolysis [lactate dehydrogenase (LDH), indirect bilirubin (IB) and reticulocyte counts], inflammation [interleukins IL-1β, IL-6, IL-8, Tumor Necrosis Factor (TNF-α) and C-reactive protein (CRP)] and endothelial dysfunction (ED) [soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble intercellular adhesion molecule-1 (sICAM-1), soluble L-selectin (sL-selectin), von Willebrand Factor (vWF) antigen and D-dimers] in the patients. The frequencies of the mutant -786C allele and -786C/C genotype were significantly higher in patients (p = 0.02 and p = 0.04, respectively) but not significantly correlated with the markers. For VNTR4a/b and G894T, the allelic/genotypic frequencies did not statistically differ between patient and control groups. Patients carrying the 4a allele and those with the 894G/G genotype showed a significant decrease in IB (p = 0.02 and p = 0.04, respectively), and only patients with the 4a allele exhibited reduced IL-1β (p = 0.01). The correlation profiles between markers of inflammation and ED significantly differed between patients carrying the mutant alleles and those with wild-type genotypes. This appears to be the first report on the relationship between eNOS gene polymorphisms and markers of hemolysis, inflammation and ED in Brazilian SCA patients. Our results indicate that the SNPs analyzed may influence the phenotypic variability of these patients.

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References

  • Armenis I, Kalotychou V, Tzanetea R, Kollia P, Kontogeorgiou Z, Anastasopoulou D, Mantzourani M, Samarkos M, Pantos K, Konstantopoulos K, Rombos I (2017) Prognostic value of T786C and G894T eNOS polymorphisms in Sickle Cell Disease. Nitric Oxide 62:17–23

    Article  CAS  Google Scholar 

  • Davis L, Dibner M, Battey J (1986) Basic Methods in Molecular Biology, 1strst edn. Elsevier, London

    Google Scholar 

  • Guzik TJ, Korbut R, Adamek-Guzik T (2003) Nitric oxide and superoxide in inflammation and immune regulation. J Physiol Pharmacol 54:469–487

    CAS  PubMed  Google Scholar 

  • Gomes Lima RA, Ribeiro Sousa V, Nicolau ID (2014) Trends in mortality and hospital admissions of sickle cell disease patients before and after the newborn screening program in Maranhão. Brazil Rev Bras Hematol Hemoter 37:12–16

    Article  Google Scholar 

  • Gombos T, Makó V, Cervenak L, Papassotiriou J, Kunde J, Hársfalvi J, Förhécz Z, Pozsonyi Z, Borgulya G, Jánoskuti L, Prohászka Z (2009) Levels of von Willebrand factor antigen and von Willebrand factor cleaving protease ( ADAMTS13) activity predict clinical events in chronic heart failure. Thromb Haemost 102:573–580

    Article  CAS  Google Scholar 

  • Holla LI, Jurajda M, Pohunek P, Znojil V (2008) Haplotype analysis of the endothelial nitric oxide synthase gene in asthma. Hum Immunol 69:306–313

    Article  CAS  Google Scholar 

  • Joshi MS, Mineo C, Shaul PW, Bauer JA (2017) Biochemical consequences of the NOS3 Glu298Asp variation in human endothelium: altered caveolar localization and impaired response to shear. FASEB J 21:2655–2663

    Article  Google Scholar 

  • Lervolino LG, Baldin Almeida EP, Picado SM, Calil KB, Viel AA, Campos Freixo AL (2011) Prevalence of sickle cell disease and sickle cell trait in national neonatal screening studies. Rev Bras Hematol Hemoter 33:49–55

    Article  Google Scholar 

  • Leonardo DP, Albuquerque DM, Lanaro C, Baptista LC, Cecatti JG, Surita FG, Parpinelli MA, Costa FF, Franco-Penteado CF, Fertrin KY, Costa ML (2015) Association of Nitric Oxide Synthase and Matrix Metalloprotease Single Nucleotide Polymorphisms with Preeclampsia and Its Complications. PLoS Negl Trop Dis 10:1–11

    Google Scholar 

  • Li R, Lyn D, Lapu-bula R, Oduwole A, Igho-pemu P, Lankford B, Morgan J, Nkemdechi S, Liu G, Pack C, Silvestrov N, Von Deutsch DA, Song Q, Abukhalaf IK, Ofili E (2004) Relation of endothelial nitric oxide synthase gene to plasma nitric oxide level, endothelial function, and blood pressure in african americans. Am J Hypertens 17:560–567

    Article  CAS  Google Scholar 

  • Marsden PA, Heng HHQ, Scherer SW, Stewart RJ, Hall AV, Shi X-M, Tsui LC, Schappert KT (1993) Structure and chromosomal localization of the human constitutive endothelial nitric oxide synthase gene. J Biol Chem 268:17478–17488

    CAS  PubMed  Google Scholar 

  • Manta FSN, Pereira R, Vianna R, Araújo ARB, Gitaí DLG, Silva DA, Wolfgramm EV, Pontes IM, Aguiar JI, Moraes MO, Carvalho EF, Gusmão L (2013) Revisiting the genetic ancestry of brazilians using autosomal AIM-Indels. PLoS ONE 8:1–11

    Google Scholar 

  • Mack AK, Kato GJ (2006) Sickle cell disease and nitric oxide: a paradigm shift? Int J Biochem Cell Biol 38:1237–1243

    Article  CAS  Google Scholar 

  • Miyamoto Y, Saito Y, Nakayama M, Shimasaki Y, Yoshimura T, Yoshimura M, Harada M, Kajiyama N, Kishimoto I, Kuwahara K, Hino J, Ogawa E, Hamanaka I, Kamitani S, Takahashi N, Kawakami R, Kangawa K, Yasue H (2000) Nakao K replication protein A1 reduces transcription of the endothelial nitric oxide synthase gene containing a –786T → C mutation associated with coronary spastic angina. Hum Mol Genet 9:2629–2637

    Article  CAS  Google Scholar 

  • Nakayama M, Yasue H, Yoshimura M, Shimasaki Y, Kugiyama K, Ogawa H, Motoyama T, Saito Y, Ogawa Y, MiyamotoY NK (1999) T-786-%3eC Mutation in the 5’-flanking region of the endothelial nitric oxide synthase gene is associated with coronary spasm. Circulation 99:2864–2870

    Article  CAS  Google Scholar 

  • Nishank SS, Singh MPSS, Yadav R, Gupta RB, Gadge VS (2013) Endothelial nitric oxide synthase gene polymorphism is associated with sickle cell disease patients in India. J Hum Genetics 58:775–779

    Article  CAS  Google Scholar 

  • Navarro KG, Agyingi SE, Nwabuobi CK, Thomas BN (2016) Polymorphism of the endothelin-1 gene ( rs5370) is a potential contributor to Sickle Cell Disease pathophysiology. Genes Dis 3:294–298

    Article  CAS  Google Scholar 

  • Reiter CD, Wang X, Tanus-Santos JE, Hogg N, Cannon RO, Schechter AN, Gladwin MT (2002) Cell-free hemoglobin limits nitric oxide bioavailability in sickle-cell disease. Nat Med 8:1383–1389

    Article  CAS  Google Scholar 

  • Sabarense AP, Lima GO, Silva LML, Viana BM (2015) Characterization of mortality in children with sickle cell disease diagnosed through the newborn screening. J Pediatr 91:242–247

    Article  Google Scholar 

  • Syed R, Biyabani MU, Prasad S, Deeba F, Jamil K (2010) Correlation and identification of variable number of tandem repeats of eNOS gene in coronary artery disease ( CAD ). Saudi J Biol Sci 17:209–213

    Article  CAS  Google Scholar 

  • Sharan K, Surrey S, Ballas S, Borowski M, Devoto M, Wang K, Sandler E, Keller M (2004) Association of T-786C eNOS gene polymorphism with increased susceptibility to acute chest syndrome in females with sickle cell disease. Br J Haematol 124:240–243

    Article  CAS  Google Scholar 

  • Sanchaisuriya K, Chunpanich S, Fucharoena G, Fucharoen S (2004) Multiplex allele-specific PCR assay for differential diagnosis of Hb S, Hb D-Punjab and Hb Tak. Clin Chim acta 343:129–134

    Article  CAS  Google Scholar 

  • Tantawy AAG, Adly AAM, Ismail EAR, Aly SH (2015) Endothelial nitric oxide synthase gene intron 4 VNTR polymorphism in sickle cell disease : relation to vasculopathy and disease severity. Pediatr Blood Cancer 62:389–394

    Article  CAS  Google Scholar 

  • Tanus-santos JE, Desai M, Flockhart DA (2001) Effects of ethnicity on the distribution of clinically relevant endothelial nitric oxide variants. Pharmacogenetics 11:719–725

    Article  CAS  Google Scholar 

  • Thakur TJ, Guindo A, Cullifer LR, Li Y, Imumorin IG, Diallo DA, Thomas BN (2014) Nitric Endothelial Synthase Oxide Polymorphism Gene. Gene Regul Syst Bio 8:119–126

    PubMed  PubMed Central  Google Scholar 

  • Vekilov PG (2007) Sickle-cell haemoglobin polymerization: is it the primary pathogenic event of sickle-cell anaemia? Br J Haematol 39:173–184

    Article  Google Scholar 

  • Vilas-boas W, Figueiredo CVB, Pitanga TN, Carvalho MOS, Santiago RP, Santana SS, Guarda CC, Zanette AMD, Cerqueira BAV, Gonçalves MS (2016) Endothelial Nitric Oxide Synthase (− 786T%3eC ) and Endothelin-1 (5665G%3eT ) Gene Polymorphisms as Vascular Dysfunction Risk Factors in Sickle Cell Anemia. Gene Regul Syst Bio 1:67–72

    Google Scholar 

  • Vargas AE, Silva MAL, Silla L, Chies JAB, Chies AB (2005) Polymorphisms of chemokine receptors and eNOS in Brazilian patients with sickle cell disease. Tissue Antigens 66:683–690

    Article  CAS  Google Scholar 

  • Wang XL, Mahaney MC, Sim AS, Wang J, Wang J, Blangero J, Almasy L, Badenhop RB, Wilcken DEL (1997) Genetic Contribution of the endothelial constitutive nitric oxide synthase gene to plasma nitric oxide levels. Arter Thromb Vasc Biol 17:3147–3153

    Article  CAS  Google Scholar 

  • Yousry SM, Ellithy HN, Shahin GH (2016) Sickle Cell Disease Endothelial nitric oxide synthase gene polymorphisms and the risk of vasculopathy in sickle cell disease. Hematology 5332:1–9

    Google Scholar 

  • Yoshimura M, Yasue H, Nakayama M, Shimasaki Y, Sumida H, Sugiyama S, Kugiyama K, Ogawa H, Ogawa Y, Saito Y, Miyamoto Y, Nakao K (1998) A missense Glu298Asp variant in the endothelial nitric oxide synthase gene is associated with coronary spasm in the Japanese. Hum Genet 103:65–69

    Article  CAS  Google Scholar 

  • Zhang M, Ou H, Shen YH, Wang J, Wang J, Coselli J, Wang XL (2005) Regulation of endothelial nitric oxide synthase by small RNA. PNAS 102:16967–16972

    Article  CAS  Google Scholar 

  • Zhang M, Zhang C, Shen YH, Wang J, Li X, Chen L, Zhang Y, Coselli JS, Wang XL (2008) Effect of 27nt Small RNA on Endothelial Nitric-Oxide Synthase Expression. Mol Biol cell 19:3997–4005

    Article  CAS  Google Scholar 

Download references

Funding

This study was financially supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) (Grant No. 2014/00984-3), the Agency for the Support and Evaluation of Graduate Education (CAPES) and Fundo de Apoio ao Ensino, Pesquisa e Extensão-FAEPEX / UNICAMP (Brazil).

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Authors and Affiliations

  1. Department of Clinical Pathology - School of Medical Sciences, State University of Campinas (UNICAMP), Rua Vital Brasil 50, Campinas, SP, 13083-888, Brazil

    F. Chenou, M. H. S. L. Blotta, M. F. Sonati & M. N. N. Santos

  2. Hematology and Hemotherapy Center, State University of Campinas-UNICAMP, Campinas, São Paulo, Brazil

    D. M. Albuquerque, D. P. Leonardo, F. F. Costa & E. V. Paula

  3. Department of Genetics, Federal University of Pernambuco (UFPE), Recife, Pernambuco, Brazil

    I. F. Domingos & M. A. C. Bezerra

  4. Hematology and Hemotherapy Foundation of Pernambuco (HEMOPE), Recife, Pernambuco, Brazil

    A. S. Araújo

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  1. F. Chenou
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  2. D. M. Albuquerque
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  3. D. P. Leonardo
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  4. I. F. Domingos
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  5. M. A. C. Bezerra
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Correspondence to M. N. N. Santos.

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All authors declare that they have no conflicts of interest with the contents of this article.

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All procedures followed complied with the ethical standards of the HEMOPE and UNICAMP committee on human research and the Helsinki Declaration of 1975, as revised in 2008. The patients and voluntary controls gave their written informed consent for this study.

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Chenou, F., Albuquerque, D.M., Leonardo, D.P. et al. Endothelial Nitric Oxide Synthase (eNOS) Gene Polymorphisms and Markers of Hemolysis, Inflammation and Endothelial Dysfunction in Brazilian Sickle Cell Anemia Patients. Biochem Genet 58, 580–594 (2020). https://doi.org/10.1007/s10528-020-09959-w

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  • DOI: https://doi.org/10.1007/s10528-020-09959-w

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