Development of a Gene and Oligonucleotide Delivery System

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 DESCRIPTION (provided by applicant): The knockdown of targeted genes by anti-sense oligonucleotides (ODNS) and genetic medicines (collectively, G-MEDS) holds promise for a variety of therapies. The delivery of effective quantities of ODNS to specific cells, however, has proved to be challenging. We propose here a novel approach to ODN delivery that involves enveloped virus-like-particles (EVLPs). These delivery agents are prepared by the in vitro self- assembly of pure G-MEDS and pure viral capsid protein (CP) into virus-like nanoparticles particles (VLPs). The capsid protects the contents yet, as we have demonstrated, is capable of giving up its contents in the cytoplasm of mammalian cells. The particles, which are highly mono-disperse, are then enveloped by lipid bilayers that can suppress the immunogenicity of the VLPs and are capable of being functionalized for targeting and uptake by mammalian cells of interest. In preliminary experiments, we have demonstrated our ability to prepare EVLPs using the CP of the cowpea chlorotic mottle virus and a model antisense ODN for vascular endothelial growth factor (VEGF), a protein over-expressed in many cancer cells to stimulate angiogenesis a facilitate tumor survival under low- oxygen conditions. We propose to optimize the assembly and to functionalize the lipid bilayers with epidermal growth factor (EGF), which binds the EGF receptor that is over-expressed on cancer cells, especially those of breast cancer. The effectiveness of the EVLP will be demonstrated by assaying the reduction in the secretion of VEGF in cultured breast cancer cells.