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02573 DEVELOPING AN HIV MICROBICIDE BASED ON RNA INTERFERENCE Lieberman, Judy Sexual transmission of HIV occurs when cell-free or cell-associated virus infects cells primarily via the CCR5 coreceptor, expressed on macrophages, dendritic cells and activated T lymphocytes. A microbicide that could be used vaginally to prevent sexual transmission would make a substantial contribution to controlling the spread of HIV. We are exploring the hypothesis that RNA interference (RNAi) can form the basis of an effective anti-HIV microbicide. RNAi is an ancient, evolutionarily conserved, host defense against viruses and transposable elements, which uses small double-stranded RNAs, called small interfering RNAs (siRNA), to silence gene expression with exquisite specificity by targeted degradation of homologous mRNAs. There has been a lot of excitement about the therapeutic potential of RNAi to treat viral infection. A major obstacle is how to deliver siRNAs into cells in vivo. Duplex siRNAs targeting CCR5 and HIV gag delivered to monocyte-derived macrophages and activated T cells lead to prolonged gene silencing, lasting for weeks in macrophages, that completely inhibits de novo infection and suppresses viral replication in already infected cells. This suggests that duplex siRNAs might serve as the active component in a microbicide that might not need to be administered directly before sexual intercourse. Many steps are needed to determine whether an siRNA-based microbicide is possible. These include in vivo delivery of siRNAs to dendritic cells, macrophages, and (if possible) lymphocytes in the genital mucosa of small animals, and demonstration that delivered siRNAs effectively inhibit HIV production. Delivery methods have to be safe and compatible with a formulation suitable for vaginal delivery that does not induce inflammation at the mucosa. Early proof-of-principle studies eventually need to be complemented by formal pharmacokinetics, toxicity and efficacy studies in small animals and primates. We will discuss our first steps to develop methods to deliver duplex siRNA to macrophages and dendritic cells in vivo in mice. Dr Judy Lieberman |
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