Dissection of experimental asthma with DNA microarray analysis identifies arginase in asthma pathogenesis
Nives Zimmermann, … , Qutayba Hamid, Marc E. Rothenberg
Nives Zimmermann, … , Qutayba Hamid, Marc E. Rothenberg
Published June 15, 2003
Citation Information: J Clin Invest. 2003;111(12):1863-1874. https://doi.org/10.1172/JCI17912.
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Dissection of experimental asthma with DNA microarray analysis identifies arginase in asthma pathogenesis

Abstract

Asthma is on the rise despite intense, ongoing research underscoring the need for new scientific inquiry. In an effort to provide unbiased insight into disease pathogenesis, we took an approach involving expression profiling of lung tissue from mice with experimental asthma. Employing asthma models induced by different allergens and protocols, we identified 6.5% of the tested genome whose expression was altered in an asthmatic lung. Notably, two phenotypically similar models of experimental asthma were shown to have distinct transcript profiles. Genes related to metabolism of basic amino acids, specifically the cationic amino acid transporter 2, arginase I, and arginase II, were particularly prominent among the asthma signature genes. In situ hybridization demonstrated marked staining of arginase I, predominantly in submucosal inflammatory lesions. Arginase activity was increased in allergen-challenged lungs, as demonstrated by increased enzyme activity, and increased levels of putrescine, a downstream product. Lung arginase activity and mRNA expression were strongly induced by IL-4 and IL-13, and were differentially dependent on signal transducer and activator of transcription 6. Analysis of patients with asthma supported the importance of this pathway in human disease. Based on the ability of arginase to regulate generation of NO, polyamines, and collagen, these results provide a basis for pharmacologically targeting arginine metabolism in allergic disorders.

Authors

Nives Zimmermann, Nina E. King, Johanne Laporte, Ming Yang, Anil Mishra, Sam M. Pope, Emily E. Muntel, David P. Witte, Anthony A. Pegg, Paul S. Foster, Qutayba Hamid, Marc E. Rothenberg

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Figure 3

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Northern blot and arginase activity analysis. In a, Northern blot analys...
Northern blot and arginase activity analysis. In a, Northern blot analysis of arginase I and arginase II expression after OVA challenge is shown. Time points are as follows: 3H = 1 challenge, 3 hours; 18H = 1 challenge, 18 hours; 2C = 2 challenges, 18 hours. The EtBr–stained gel is also shown. The autoradiograph exposure times were 18 hours and 2 days for arginase I and arginase II, respectively. In b, the expression of arginase I and arginase II after intranasal challenges with A. fumigatus or saline is shown. Sal, saline; Asp, aspergillus. The autoradiograph exposure times were 18 hours and 6 days for arginase I and arginase II, respectively. In a and b, each lane represents a separate mouse. In c, arginase activity in the lungs of saline- and OVA-challenged mice (n = 4 mice and n = 3 mice, respectively) is shown. Arginase activity was measured in lung lysates with the use of the blood urea nitrogen reagent. As a control, arginase activity in the liver was 1522 ± 183 and 1390 ± 78 nmol/min/mg protein for saline- and OVA-challenged mice, respectively. In d, putrescine levels in the whole lungs of saline- and OVA-challenged mice (n = 4 mice and n = 7 mice, respectively) are shown. Putrescine levels were determined by HPLC.