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High-throughput gene expression analysis to screen for anti-asthma drugs

Andreas Pahl, Uwe Oelmller,* and Susanne Ullmann*
Department of Pharmacology, University of Erlangen, Erlangen, Germany

* QIAGEN GmbH, Hilden Germany Drug library screening based on gene expression analysis requires reliable and reproducible methods of RNA purification and sensitive, quantitative, real-time RT-PCR analysis. This article demonstrates how RNeasy 96 technology, automated on the BioRobot Gene Expression workstation, enabled reproducible, high-throughput sample preparation. One-step, real-time RT-PCR using the QuantiTect Probe RT-PCR Kit provided highly sensitive gene expression analysis to identify potential drugs to fight asthma.

Bronchial asthma is one of the commonest chronic diseases in modern society and the most common chronic childhood disease. The pathogenesis and causes of asthma are still poorly understood. Nonetheless, our knowledge of asthma pathogenesis has increased greatly over the past two decades, partially through recognition of the critical role played by cytokines in the inflammatory response.

Many cytokines are involved in asthma. Although some of these cytokines are involved in many inflammatory diseases, others are more specific to allergic inflammation, such as asthma. These cytokines include interleukins (IL) IL-4 and IL-13, which are derived from T-helper 2 (Th2) cells. Th1 cells, which are involved in cellular immunity, secrete other cytokines, such as IL-2. Gene expression analysis of these cytokines provides valuable information about the effects of potential drugs for asthma.

The goal of our research is to identify potential new drugs that selectively
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