In vitro-targeted gene identification in patients with hepatitis C using a genome-wide microarray.pdfVIP

In Vitro–Targeted Gene Identification in Patients with Hepatitis C Using a Genome-Wide Microarray Technology Susanne Hagist,1 ¨ 2 1 1 ¨ 1 2 * Holger Sultmann, * Gunda Millonig, Ulrike Hebling, Dorthe Kieslich, Rupert Kuner, Sabrina Balaguer,2 1 2 1 Helmut-Karl Seitz, Annemarie Poustka, and Sebastian Mueller Iron in association with reactive oxygen species (ROS) is highly toxic, aggravating oxidative stress reactions. Increased iron not only plays an important role in the progression of hereditary hemochromatosis (HH) but also in common liver diseases such as chronic hepa- titis C. The underlying mechanisms of hepatitis C virus (HCV)-mediated iron accumulation, however, are poorly understood. We introduce an in vitro–targeted approach to identify ROS/iron-regulated genes in patients with HCV using a genome-wide DNA microarray. The sensitivity of the 32,231 complementary DNA clone-carrying microarray was approximately 20% as estimated by detecting target genes of the genome-wide transcription factor hypoxia inducible factor 1. Upon in vitro challenge to iron and oxidative stress, 265 iron-related and 1326 ROS-related genes could be identified in HepG2 cells; 233 significantly regulated genes were found in patients with mild (HCV) or severe (HH) iron deposition. Notably, 17 of the in vitro–selected genes corresponded to the genes identified in patients with HCV or