title: The targeted intracellular delivery of cytochrome C protein to tumors using lipid-apolipoprotein nanoparticles.
abstract: Intracellular-acting therapeutic proteins offer a promising clinical alternative to extracellular-acting agents, but are limited in efficacy by their low permeability into the cell cytoplasm. We have developed a nanoparticle (NP) composed of lipid (DOTAP/DOPE) and apolipoprotein (APOA-I) to mediate the targeted delivery of intracellular-acting protein drugs to non-small cell lung tumors. NPs were produced with either GFP, a fluorescent model protein, or cytochrome C (cytC), an inducer of apoptosis in cancer cells. GFP and cytC were separately conjugated with a membrane permeable sequence (MPS) peptide and were admixed with DOPE/DOTAP nanoparticle formulations to enable successful protein loading. Protein-loaded NPs were modified with DSPE-PEG-Anisamide to enable specific NP targeting to the tumor site in a xenograft model. The resulting particle was 20-30 nm in size and exhibited a 64-75% loading efficiency. H460 cells treated with the PEGylated MPS-cytC-NPs exhibited massive apoptosis. When MPS-GFP-NPs or MPS-cytC-NPs were intravenously administered in H460 tumor bearing mice, a specific tumor targeting effect with low NP accumulation in the liver was observed. In addition, MPS-cytC-NP treatment provoked a tumor growth retardation effect in H460 xenograft mice. We conclude that our NP enables targeted, efficacious therapeutic protein delivery for the treatment of lung cancer.
authors.affiliation: Division of Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC 27599, USA.
authors.forename: Sang Kyoon
authors.initials: SK
authors.lastname: Kim
authors.forename: Michael B
authors.initials: MB
authors.lastname: Foote
authors.forename: Leaf
authors.initials: L
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D001053:Apolipoproteins / Q000737:chemistry*; D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D049109:Cell Proliferation; D045304:Cytochromes c / Q000627:therapeutic use*; D016503:Drug Delivery Systems / Q000379:methods*; D004789:Enzyme Activation; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000008:administration & dosage / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007275:Injections, Intravenous; D042541:Intracellular Space / Q000378:metabolism*; D008055:Lipids / Q000737:chemistry*; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D009369:Neoplasms / Q000188:drug therapy* / Q000473:pathology; D010316:Particle Size; D011419:Propidium / Q000378:metabolism; D014018:Tissue Distribution; D023041:Xenograft Model Antitumor Assays
chemical_list: D017304:Annexin A5; D001053:Apolipoproteins; D008055:Lipids; D049452:Green Fluorescent Proteins; D011419:Propidium; D045304:Cytochromes c; D053148:Caspase 3