Abstract
Genetic polymorphisms in superoxide-producing NAD(P)H oxidase have been linked to cardiovascular diseases including anthracycline-induced cardiotoxicity. We quantified NAD(P)H oxidase activity in granulocytes of 81 healthy Caucasian volunteers (in addition, 51 in an independent confirmatory study) by chemiluminescence using the luminol analogue L-012. Expression of CYBA, NCF4 and RAC2 coding for NAD(P)H oxidase subunits was measured in whole blood cells in 59 study participants by real-time PCR. Of the five variants investigated (−930A>G, 242C>T, 640A>G in CYBA and the recently reported −368G>A in NCF4 and 7508T>A in RAC2), only CYBA 640A>G was consistently associated with superoxide production (640GG carriers 28% less than AA individuals, P=0.05 in each cohort, P=0.005 in combined analysis). RAC2 7508T>A was related to higher expression of RAC2 (P=0.02) and NCF4 (P=0.04). In summary, CYBA 640A>G rather than 242C>T was associated with reduced activity. The quantitatively moderate effect and the high intra-individual variability should be considered for further study design.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 6 print issues and online access
$259.00 per year
only $43.17 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Babior BM . Oxygen-dependent microbial killing by phagocytes (first of two parts). N Engl J Med 1978; 298: 659–668.
Babior BM . Oxygen-dependent microbial killing by phagocytes (second of two parts). N Engl J Med 1978; 298: 721–725.
Sauer H, Wartenberg M, Hescheler J . Reactive oxygen species as intracellular messengers during cell growth and differentiation. Cell Physiol Biochem 2001; 11: 173–186.
Shen WL, Gao PJ, Che ZQ, Ji KD, Yin M, Yan C et al. NAD(P)H oxidase-derived reactive oxygen species regulate angiotensin-II induced adventitial fibroblast phenotypic differentiation. Biochem Biophys Res Commun 2006; 339: 337–343.
Pervaiz S, Clement MV . A permissive apoptotic environment: function of a decrease in intracellular superoxide anion and cytosolic acidification. Biochem Biophys Res Commun 2002; 290: 1145–1150.
Pervaiz S, Clement MV . Hydrogen peroxide-induced apoptosis: oxidative or reductive stress? Methods Enzymol 2002; 352: 150–159.
Sato A, Sakuma I, Gutterman DD . Mechanism of dilation to reactive oxygen species in human coronary arterioles. Am J Physiol Heart Circ Physiol 2003; 285: H2345–H2354.
Tabner BJ, Turnbull S, Fullwood NJ, German M, Allsop D . The production of hydrogen peroxide during early-stage protein aggregation: a common pathological mechanism in different neurodegenerative diseases? Biochem Soc Trans 2005; 33: 548–550.
Ferraro D, Corso S, Fasano E, Panieri E, Santangelo R, Borrello S et al. Pro-metastatic signaling by c-Met through RAC-1 and reactive oxygen species (ROS). Oncogene 2006.
Mitsushita J, Lambeth JD, Kamata T . The superoxide-generating oxidase Nox1 is functionally required for Ras oncogene transformation. Cancer Res 2004; 64: 3580–3585.
Kinugawa S, Post H, Kaminski PM, Zhang X, Xu X, Huang H et al. Coronary microvascular endothelial stunning after acute pressure overload in the conscious dog is caused by oxidant processes: the role of angiotensin II type 1 receptor and NAD(P)H oxidase. Circulation 2003; 108: 2934–2940.
Lambeth JD . NOX enzymes and the biology of reactive oxygen. Nat Rev Immunol 2004; 4: 181–189.
Babior BM . NADPH oxidase. Curr Opin Immunol 2004; 16: 42–47.
Bengtsson SH, Gulluyan LM, Dusting GJ, Drummond GR . Novel isoforms of NADPH oxidase in vascular physiology and pathophysiology. Clin Exp Pharmacol Physiol 2003; 30: 849–854.
Hoyal CR, Gutierrez A, Young BM, Catz SD, Lin JH, Tsichlis PN et al. Modulation of p47PHOX activity by site-specific phosphorylation: Akt-dependent activation of the NADPH oxidase. Proc Natl Acad Sci USA 2003; 100: 5130–5135.
Inanami O, Johnson JL, McAdara JK, Benna JE, Faust LR, Newburger PE et al. Activation of the leukocyte NADPH oxidase by phorbol ester requires the phosphorylation of p47PHOX on serine 303 or 304. J Biol Chem 1998; 273: 9539–9543.
Moreno MU, San Jose G, Orbe J, Paramo JA, Beloqui O, Diez J et al. Preliminary characterisation of the promoter of the human p22(phox) gene: identification of a new polymorphism associated with hypertension. FEBS Lett 2003; 542: 27–31.
San Jose G, Moreno MU, Olivan S, Beloqui O, Fortuno A, Diez J et al. Functional effect of the p22phox −930A/G polymorphism on p22phox expression and NADPH oxidase activity in hypertension. Hypertension 2004; 44: 163–169.
Guzik TJ, West NE, Black E, McDonald D, Ratnatunga C, Pillai R et al. Functional effect of the C242T polymorphism in the NAD(P)H oxidase p22phox gene on vascular superoxide production in atherosclerosis. Circulation 2000; 102: 1744–1747.
Wyche KE, Wang SS, Griendling KK, Dikalov SI, Austin H, Rao S et al. C242T CYBA polymorphism of the NADPH oxidase is associated with reduced respiratory burst in human neutrophils. Hypertension 2004; 43: 1246–1251.
Shimo-Nakanishi Y, Hasebe T, Suzuki A, Mochizuki H, Nomiyama T, Tanaka Y et al. Functional effects of NAD(P)H oxidase p22(phox) C242T mutation in human leukocytes and association with thrombotic cerebral infarction. Atherosclerosis 2004; 175: 109–115.
Gardemann A, Mages P, Katz N, Tillmanns H, Haberbosch W . The p22 phox A640G gene polymorphism but not the C242T gene variation is associated with coronary heart disease in younger individuals. Atherosclerosis 1999; 145: 315–323.
Wojnowski L, Kulle B, Schirmer M, Schluter G, Schmidt A, Rosenberger A et al. NAD(P)H oxidase and multidrug resistance protein genetic polymorphisms are associated with doxorubicin-induced cardiotoxicity. Circulation 2005; 112: 3754–3762.
Kalow W, Tang BK, Endrenyi L . Hypothesis: comparisons of inter- and intra-individual variations can substitute for twin studies in drug research. Pharmacogenetics 1998; 8: 283–289.
Park JY, Ferrell RE, Park JJ, Hagberg JM, Phares DA, Jones JM et al. NADPH oxidase p22phox gene variants are associated with systemic oxidative stress biomarker responses to exercise training. J Appl Physiol 2005; 99: 1905–1911.
Doi K, Noiri E, Nakao A, Fujita T, Kobayashi S, Tokunaga K . Haplotype analysis of NAD(P)H oxidase p22 phox polymorphisms in end-stage renal disease. J Hum Genet 2005; 50: 641–647.
Li SL, Valente AJ, Qiang M, Schlegel W, Gamez M, Clark RA . Multiple PU.1 sites cooperate in the regulation of p40(phox) transcription during granulocytic differentiation of myeloid cells. Blood 2002; 99: 4578–4587.
Anrather J, Racchumi G, Iadecola C . NF-kappaB regulates phagocytic NADPH oxidase by inducing the expression of gp91phox. J Biol Chem 2006; 281: 5657–5667.
Daiber A, August M, Baldus S, Wendt M, Oelze M, Sydow K et al. Measurement of NAD(P)H oxidase-derived superoxide with the luminol analogue L-012. Free Radic Biol Med 2004; 36: 101–111.
Nishinaka Y, Aramaki Y, Yoshida H, Masuya H, Sugawara T, Ichimori Y . A new sensitive chemiluminescence probe, L-012, for measuring the production of superoxide anion by cells. Biochem Biophys Res Commun 1993; 193: 554–559.
Allen RC . Phagocytic leukocyte oxygenation activities and chemiluminescence: a kinetic approach to analysis. Methods Enzymol 1986; 133: 449–493.
Inoue N, Kawashima S, Kanazawa K, Yamada S, Akita H, Yokoyama M . Polymorphism of the NADH/NADPH oxidase p22 phox gene in patients with coronary artery disease. Circulation 1998; 97: 135–137.
Acknowledgements
This work was supported by Grants 01GS0107, 01GS0157, 01GR0462 and 01GS0421 of the National Genome Research Network, Federal Ministry of Education and Research, Bonn, Germany; by the Deutsche Forschungsgemeinschaft (Graduiertenkolleg 1034), Bonn, Germany; and by the local research promotion programme of Göttingen University, Germany. We gratefully acknowledge the support of Daniel Sehrt, MD, and Stefan V Vormfelde, MD, concerning volunteer recruitment. And the authors acknowledge the technical assistance of Karoline Jobst and the skillful contributions of the medical students Daniela Bolte, Susanne Pahl, Julia P Kaup, Irmani B Ismadi and Adda Boekhoff. We are thankful to the kind support of the Department of Transfusion Medicine at Göttingen University Hospital.
Author information
Authors and Affiliations
Corresponding author
Additional information
Duality of interest
The authors have no conflicts of interest to disclose.
Rights and permissions
About this article
Cite this article
Schirmer, M., Hoffmann, M., Kaya, E. et al. Genetic polymorphisms of NAD(P)H oxidase: variation in subunit expression and enzyme activity. Pharmacogenomics J 8, 297–304 (2008). https://doi.org/10.1038/sj.tpj.6500467
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.tpj.6500467
Keywords
This article is cited by
-
Impact of NADPH oxidase functional polymorphisms in acute myeloid leukemia induction chemotherapy
The Pharmacogenomics Journal (2018)
-
The effect of the SNP g.18475 A>G in the 3′UTR of NCF4 on mastitis susceptibility in dairy cattle
Cell Stress and Chaperones (2018)
-
Association between NCF4 rs4821544T/C polymorphism and inflammatory bowel disease risk in Caucasian: a meta-analysis
Inflammation Research (2015)
-
The −930A>G polymorphism of the CYBA gene is associated with premature coronary artery disease. A case–control study and gene–risk factors interactions
Molecular Biology Reports (2014)
-
Impact of genetic variability and treatment-related factors on outcome in early breast cancer patients receiving (neo-) adjuvant chemotherapy with 5-fluorouracil, epirubicin and cyclophosphamide, and docetaxel
Breast Cancer Research and Treatment (2014)