? The ultimate purpose of this project is to discover new classes of anti-HIV (human immunodeficiency virus) drugs by identifying and validating human genes that are required for viral replication, maturation or infection. The initial research project will be pursued jointly between Oregon Health & Science University and Virogenomics, Inc., with Virogenomics and its partners responsible for continuing the drug discovery process on validated candidate genes. The strategy is to perform microarray gene expression analysis on cultured human immune cells (T cells and macrophages) at multiple time points after infection with several strains of HIV and to identify genes that are at least twofold induced or repressed relative to uninfected controls. Using the Affymetrix U133 A and B gene arrays, over 33,000 potential gene transcripts from the human genome can be assayed. Over half of these transcripts are as yet uncharacterized beyond primary sequence and represent a potentially rich source of novel targets. Based on previous experience with other viruses, up to 300 genes may qualify as initial candidates. The next step will be to test the roles of those genes in HIV infection. As a proof of principle in Phase I we will create antisense or small interfering RNA molecules specific to 20-30 upregulated genes. After administering the sequence-specific silencing molecules, the cells will be infected with HIV and the infection will be monitored visually by indirect immunofluorescence and by following p24 (HIV polymerase) activity using ELISA. If silencing the gene activity results in reduced HIV infection, replication or spread, the gene will be considered a validated candidate for further study as a drug development target. In Phase II, the project will be extended to include multiple cell lines and HIV strains as well as cells isolated directly from human blood samples. This project will be the first comprehensive study of the expression patterns of the entire human genome induced by HIV over the course of an infection, coupled with an a sequence specific validation assay for the potential of each candidate gene as an anti-HIV drug. The company intends to develop intellectual property protection for the use of novel validated genes in antiviral drug assays, and pursue the development of such drugs. ? ? ?
