Development of a Gene and Oligonucleotide Delivery System

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

PUBLIC HEALTH RELEVANCE The development of the proposed system for delivering genetic medicines such as genes and oligonucleotides would be an important medical advance Its development would facilitate the targeted delivery of genetic medicines to specific cells tissues and organs thus enabling the development of healthcare products which will exert a significant impact on the practice of medicine