Levels of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, are increased in lung, sputum, exhaled breath condensate and plasma samples from asthma patients. ADMA is metabolized primarily by dimethylarginine dimethylaminohydrolase 1 (DDAH1) and DDAH2. We determined the effect of DDAH1 overexpression on development of allergic inflammation in a mouse model of asthma. The expression of DDAH1 and DDAH2 in mouse lungs was determined by RT-quantitative PCR (qPCR). ADMA levels in bronchoalveolar lavage fluid (BALF) and serum samples were determined by mass spectrometry. Wild type and DDAH1-transgenic mice were intratracheally challenged with PBS or house dust mite (HDM). Airway inflammation was assessed by bronchoalveolar lavage (BAL) total and differential cell counts. The levels of IgE and IgG1 in BALF and serum samples were determined by ELISA. Gene expression in lungs was determined by RNA-Seq and RT-qPCR. Our data showed that the expression of DDAH1 and DDAH2 was decreased in the lungs of mice following HDM exposure, which correlated with increased ADMA levels in BALF and serum. Transgenic overexpression of DDAH1 resulted in decreased BAL total cell and eosinophil numbers following HDM exposure. Total IgE levels in BALF and serum were decreased in HDM-exposed DDAH1-transgenic mice compared to HDM-exposed wild type mice. RNA-Seq results showed downregulation of genes in the inducible nitric oxide synthase (iNOS) signaling pathway in PBS-treated DDAH1-transgenic mice versus PBS-treated wild type mice and downregulation of genes in IL-13/FOXA2 signaling pathway in HDM-treated DDAH1-transgenic mice versus HDM-treated wild type mice. Our findings suggest that decreased expression of DDAH1 and DDAH2 in the lungs may contribute to allergic asthma and overexpression of DDAH1 attenuates allergen-induced airway inflammation through modulation of Th2 responses.
Malerba G, Pignatti PF (2005) A review of asthma genetics: gene expression studies and recent candidates. J Appl Genet 46: 93–104.
[3]
Lara A, Khatri SB, Wang Z, Comhair SA, Xu W, et al. (2008) Alterations of the arginine metabolome in asthma. Am J Respir Crit Care Med 178: 673–681.
[4]
Zimmermann N, Rothenberg ME (2006) The arginine-arginase balance in asthma and lung inflammation. Eur J Pharmacol 533: 253–262.
[5]
Ricciardolo FL, Sterk PJ, Gaston B, Folkerts G (2004) Nitric oxide in health and disease of the respiratory system. Physiol Rev 84: 731–765.
[6]
Vallance P, Leone A, Calver A, Collier J, Moncada S (1992) Accumulation of an endogenous inhibitor of nitric oxide synthesis in chronic renal failure. Lancet 339: 572–575.
[7]
Schmidt K, Klatt P, Mayer B (1993) Characterization of endothelial cell amino acid transport systems involved in the actions of nitric oxide synthase inhibitors. Mol Pharmacol 44: 615–621.
[8]
Bogle RG, Moncada S, Pearson JD, Mann GE (1992) Identification of inhibitors of nitric oxide synthase that do not interact with the endothelial cell L-arginine transporter. Br J Pharmacol 105: 768–770.
[9]
Bulau P, Zakrzewicz D, Kitowska K, Leiper J, Gunther A, et al. (2007) Analysis of methylarginine metabolism in the cardiovascular system identifies the lung as a major source of ADMA. Am J Physiol Lung Cell Mol Physiol 292: L18–24.
[10]
Smith CL, Anthony S, Hubank M, Leiper JM, Vallance P (2005) Effects of ADMA upon gene expression: an insight into the pathophysiological significance of raised plasma ADMA. PLoS Med 2: e264.
[11]
Wells SM, Buford MC, Migliaccio CT, Holian A (2009) Elevated asymmetric dimethylarginine alters lung function and induces collagen deposition in mice. Am J Respir Cell Mol Biol 40: 179–188.
[12]
Klein E, Weigel J, Buford MC, Holian A, Wells SM (2010) Asymmetric Dimethylarginine Potentiates Lung Inflammation in a Mouse Model of Allergic Asthma. Am J Physiol Lung Cell Mol Physiol.
[13]
Sott JA, North ML, Rafii M, Huang H, Pencharz P, et al. (2011) Asymmetric dimethylarginine is increased in asthma. Am J Respir Crit Care Med 184: 779–785.
[14]
Di Gangi IM, Pirillo P, Carraro S, Gucciardi A, Naturale M, et al. (2012) Online trapping and enrichment ultra performance liquid chromatography-tandem mass spectrometry method for sensitive measurement of “arginine-asymmetric dimethylarginine cycle” biomarkers in human exhaled breath condensate. Anal Chim Acta 754: 67–74.
[15]
Carraro S, Giordano G, Piacentini G, Kantar A, Moser S, et al.. (2013) Asymmetric Dimethylarginine (Adma) in Exhaled Breath Condensate and Serum of Asthmatic Children. Chest.
[16]
Holguin F, Comhair SA, Hazen SL, Powers RW, Khatri SS, et al. (2013) An association between L-arginine/asymmetric dimethyl arginine balance, obesity, and the age of asthma onset phenotype. Am J Respir Crit Care Med 187: 153–159.
[17]
Palm F, Onozato ML, Luo Z, Wilcox CS (2007) Dimethylarginine dimethylaminohydrolase (DDAH): expression, regulation, and function in the cardiovascular and renal systems. Am J Physiol Heart Circ Physiol 293: H3227–3245.
[18]
Kimoto M, Miyatake S, Sasagawa T, Yamashita H, Okita M, et al. (1998) Purification, cDNA cloning and expression of human NG,NG-dimethylarginine dimethylaminohydrolase. Eur J Biochem 258: 863–868.
[19]
Leiper JM, Santa Maria J, Chubb A, MacAllister RJ, Charles IG, et al. (1999) Identification of two human dimethylarginine dimethylaminohydrolases with distinct tissue distributions and homology with microbial arginine deiminases. Biochem J 343 Pt 1: 209–214.
[20]
Tran CT, Fox MF, Vallance P, Leiper JM (2000) Chromosomal localization, gene structure, and expression pattern of DDAH1: comparison with DDAH2 and implications for evolutionary origins. Genomics 68: 101–105.
[21]
Pullamsetti S, Kiss L, Ghofrani HA, Voswinckel R, Haredza P, et al. (2005) Increased levels and reduced catabolism of asymmetric and symmetric dimethylarginines in pulmonary hypertension. FASEB J 19: 1175–1177.
[22]
Leiper J, Nandi M, Torondel B, Murray-Rust J, Malaki M, et al. (2007) Disruption of methylarginine metabolism impairs vascular homeostasis. Nat Med 13: 198–203.
[23]
Hu X, Atzler D, Xu X, Zhang P, Guo H, et al. (2011) Dimethylarginine dimethylaminohydrolase-1 is the critical enzyme for degrading the cardiovascular risk factor asymmetrical dimethylarginine. Arterioscler Thromb Vasc Biol 31: 1540–1546.
Jacobi J, Sydow K, von Degenfeld G, Zhang Y, Dayoub H, et al. (2005) Overexpression of dimethylarginine dimethylaminohydrolase reduces tissue asymmetric dimethylarginine levels and enhances angiogenesis. Circulation 111: 1431–1438.
[26]
Dayoub H, Rodionov RN, Lynch C, Cooke JP, Arning E, et al. (2008) Overexpression of dimethylarginine dimethylaminohydrolase inhibits asymmetric dimethylarginine-induced endothelial dysfunction in the cerebral circulation. Stroke 39: 180–184.
[27]
Lee GB, Brandt EB, Xiao C, Gibson AM, Le Cras TD, et al. (2013) Diesel exhaust particles induce cysteine oxidation and s-glutathionylation in house dust mite induced murine asthma. PLoS One 8: e60632.
[28]
Brandt EB, Kovacic MB, Lee GB, Gibson AM, Acciani TH, et al.. (2013) Diesel exhaust particle induction of IL-17A contributes to severe asthma. J Allergy Clin Immunol. 2013 Sep 20. doi:pii: S0091-6749(13)01208-6. 10.1016/j.jaci.2013.06.048. [Epub ahead of print].
[29]
Kramer EL, Mushaben EM, Pastura PA, Acciani TH, Deutsch GH, et al. (2009) Early growth response-1 suppresses epidermal growth factor receptor-mediated airway hyperresponsiveness and lung remodeling in mice. Am J Respir Cell Mol Biol 41: 415–425.
[30]
Chen W, Ericksen MB, Levin LS, Khurana Hershey GK (2004) Functional effect of the R110Q IL13 genetic variant alone and in combination with IL4RA genetic variants. J Allergy Clin Immunol 114: 553–560.
[31]
Sivaprasad U, Warrier MR, Gibson AM, Chen W, Tabata Y, et al. (2010) IL-13Ralpha2 has a protective role in a mouse model of cutaneous inflammation. J Immunol 185: 6802–6808.
Morgan M, Anders S, Lawrence M, Aboyoun P, Pages H, et al. (2009) ShortRead: a bioconductor package for input, quality assessment and exploration of high-throughput sequence data. Bioinformatics 25: 2607–2608.
[34]
Ihaka R, Gentleman R (1996) R: A Language for Data Analysis and Graphics. J Comput Graph Stat 5: 299–314.
[35]
Anders S, Huber W (2010) Differential expression analysis for sequence count data. Genome Biol 11: R106.
[36]
Storey JD, Tibshirani R (2003) Statistical significance for genomewide studies. Proc Natl Acad Sci U S A 100: 9440–9445.
[37]
Sartor MA, Leikauf GD, Medvedovic M (2009) LRpath: a logistic regression approach for identifying enriched biological groups in gene expression data. Bioinformatics 25: 211–217.
[38]
Schwedhelm E, Tan-Andresen J, Maas R, Riederer U, Schulze F, et al. (2005) Liquid chromatography-tandem mass spectrometry method for the analysis of asymmetric dimethylarginine in human plasma. Clin Chem 51: 1268–1271.
[39]
Tomikawa J, Fukatsu K, Tanaka S, Shiota K (2006) DNA methylation-dependent epigenetic regulation of dimethylarginine dimethylaminohydrolase 2 gene in trophoblast cell lineage. J Biol Chem 281: 12163–12169.
[40]
Sujuan Y, Asaithambi A, Liu Y (2008) CpGIF: an algorithm for the identification of CpG islands. Bioinformation 2: 335–338.
[41]
Kwon NH, Kim JS, Lee JY, Oh MJ, Choi DC (2008) DNA methylation and the expression of IL-4 and IFN-gamma promoter genes in patients with bronchial asthma. J Clin Immunol 28: 139–146.
[42]
Kelada SN, Wilson MS, Tavarez U, Kubalanza K, Borate B, et al. (2011) Strain-dependent genomic factors affect allergen-induced airway hyperresponsiveness in mice. Am J Respir Cell Mol Biol 45: 817–824.
[43]
Zhu Z, Zheng T, Homer RJ, Kim YK, Chen NY, et al. (2004) Acidic mammalian chitinase in asthmatic Th2 inflammation and IL-13 pathway activation. Science 304: 1678–1682.
[44]
Chang NC, Hung SI, Hwa KY, Kato I, Chen JE, et al. (2001) A macrophage protein, Ym1, transiently expressed during inflammation is a novel mammalian lectin. J Biol Chem 276: 17497–17506.
[45]
Webb DC, McKenzie AN, Foster PS (2001) Expression of the Ym2 lectin-binding protein is dependent on interleukin (IL)-4 and IL-13 signal transduction: identification of a novel allergy-associated protein. J Biol Chem 276: 41969–41976.
[46]
Welch JS, Escoubet-Lozach L, Sykes DB, Liddiard K, Greaves DR, et al. (2002) TH2 cytokines and allergic challenge induce Ym1 expression in macrophages by a STAT6-dependent mechanism. J Biol Chem 277: 42821–42829.