Development of a genetically engineered biomimetic vector for targeted gene transfer to breast cancer cells.
A biomimetic vector was genetically engineered to contain at precise locations (a) an adenovirus mu peptide to condense pDNA into nanosize particles, (b) a synthetic cyclic peptide to target breast cancer cells and enhance internalization of nanoparticles, (c) a pH-responsive synthetic fusogenic peptide to disrupt endosome membranes and facilitate escape of the nanoparticles into the cytosol, and (d) a nuclear localization signal from human immunodeficiency virus for microtubule mediated transfer of genetic material to the nucleus. The vector was characterized using physicochemical and biological assays to demonstrate the functionality of each motif in the vector backbone. The results demonstrated that the vector is able to condense plasmid DNA into nanosize particles (<100 nm), protect pDNA from serum endonucleases, target ZR-75-1 breast cancer cells and internalize, efficiently disrupt endosome membranes, exploit microtubules to reach nucleus and mediate gene expression. The therapeutic potential of the vector was evaluated by complexing with plasmid DNA encoding TRAIL (pTRAIL) and transfecting ZR-75-1 cells. The results demonstrated that up to 62% of the ZR-75-1 breast cancer cells can be killed after administration of pTRAIL in complex with the vector.
Department of Pharmaceutical Sciences, Center for Integrated Biotechnology, Washington State University, Pullman, Washington 99164, USA.
Sriramchandra S
SS
Mangipudi
Brenda F
BF
Canine
Yuhua
Y
Wang
Arash
A
Hatefi
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017209:Apoptosis; D032701:Biomimetics; D001943:Breast Neoplasms / Q000235:genetics / Q000628:therapy*; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000008:administration & dosage*; D011992:Endosomes / Q000378:metabolism; D005260:Female; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000627:therapeutic use*; D006461:Hemolysis; D006801:Humans; D008870:Microtubules / Q000378:metabolism; D053758:Nanoparticles; D036103:Nanotechnology; D010446:Peptide Fragments / Q000378:metabolism; D012756:Sheep; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured
D010446:Peptide Fragments; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; D004247:DNA