Toward a siRNA-containing nanoparticle targeted to breast cancer cells and the tumor microenvironment

• Published in: International journal of pharmaceutics 2012-09, Vol.434 (1-2), p.9-19
• Main Authors: Gomes-da-Silva, Lígia C., Santos, Adriana O., Bimbo, Luís M., Moura, Vera, Ramalho, José S., Pedroso de Lima, Maria C., Simões, Sérgio, Moreira, João N.
• Format: Article
• Language: English
• Subjects: blood vessels; Breast cancer; breast neoplasms; Breast Neoplasms - blood supply; Breast Neoplasms - genetics; Breast Neoplasms - therapy; Cell Line; Cell Line, Tumor; Down-Regulation; Dual-targeted delivery; endothelial cells; Endothelial Cells - metabolism; ethylene glycol; Female; gene expression regulation; gene overexpression; green fluorescent protein; Green Fluorescent Proteins - genetics; Humans; Ligand-mediated targeting; Liposomes; lysosomes; Lysosomes - metabolism; messenger RNA; nanocarriers; Nanoparticles; Neovascularization, Pathologic - pathology; Neovascularization, Pathologic - therapy; Nucleolin; Peptides - metabolism; Phosphoproteins - metabolism; Polyethylene Glycols - chemistry; RNA, Messenger - metabolism; RNA, Small Interfering - administration & dosage; RNA-Binding Proteins - metabolism; siRNA; small interfering RNA; Stable nucleic acid lipid particles (SNALP); Tumor Microenvironment
• ISSN: 0378-5173; 1873-3476
• DOI: 10.1016/j.ijpharm.2012.05.018

The present work aimed at designing a lipid-based nanocarrier for siRNA delivery toward two cell sub-populations within breast tumors, the cancer and the endothelial cells from angiogenic tumor blood vessels. To achieve such goal, the F3 peptide, which is specifically internalized by nucleolin overexpressed on both those sub-populations, was used as a targeting moiety. The developed F3-targeted stable nucleic acid lipid particles presented adequate features for systemic administration. In addition, the attachment of the F3 peptide onto the liposomal surface enabled an internalization by both cancer and endothelial cells from angiogenic blood vessels that was significantly higher than the one observed with non-cancer cells. Sequence-specific downregulation of enhanced green fluorescent protein (eGFP) in eGFP-overexpressing human cancer cell lines, both at the protein and mRNA levels, was further observed upon delivery of anti-eGFP siRNA by F3-targeted liposomes, in contrast with the non-targeted counterpart. This effect was highly dependent on the content of poly(ethylene glycol) (PEG), as evidenced by the co-localization studies between the siRNA and the lysosomes. Overall, the present work represents an important contribution toward a nanoparticle with multi-targeting capabilities in breast cancer, both at the cellular and molecular level.