title: Development of a novel nanoparticle by dual modification with the pluripotential cell-penetrating peptide PepFect6 for cellular uptake, endosomal escape, and decondensation of an siRNA core complex.
abstract: Development of novel devices for effective nucleotide release from nanoparticles is required to improve the functionality of nonviral delivery systems, because decondensation of nucleotide/polycation complexes is considered as a key step for cytoplasmic delivery of nucleotides. Previously, PepFect6 (PF6) comprised chloroquine analog moieties and a stearylated cell-penetrating peptide to facilitate endosomal escape and cellular uptake, respectively, was developed as a device for efficient siRNA delivery. As PF6 contains bulky chloroquine analog moieties, the polyplexes are expected to be loose structure, which facilitates decondensation. In the present study, siRNA was electrostatically condensed by PF6, and the PF6/siRNA complexes were coated with lipid membranes. The surface of the nanoparticles encapsulating the PF6/siRNA core (PF6-NP) was modified with PF6 for endosomal escape (PF6/PF6-NP). The RNAi effect of PF6/PF6-NP was compared with those of stearylated cell-penetrating peptide octaarginine (R8)-modified PF6-NP, R8-modified nanoparticles encapsulating the R8/siRNA core (R8-NP) and PF6-modified R8-NP. Nanoparticles encapsulating the PF6 polyplex, especially PF/PF-NP, showed a significant knockdown effect on luciferase activity of B16-F1 cells stably expressing luciferase. siRNA was widely distributed within the cytoplasm after transfection of the nanoparticles encapsulating the PF6 polyplex, while siRNA encapsulated in the R8-presenting nanoparticles was localized within the nuclei. Thus, the siRNA distribution was dependent on the manner of peptide-modification. In conclusion, we have successfully developed PF6/PF6-NP exhibiting a potent RNAi effect resulting from high cellular uptake, efficient endosomal escape and decondensation of the polyplexes based on the multifunctional cell penetrating peptide PF6. PF6 is therefore a useful pluripotential device for siRNA delivery.
authors.affiliation: Department of Biophysical Chemistry, Kyoto Pharmaceutical University, Kyoto, Kyoto, 607-8414, Japan.
authors.forename: Asako
authors.initials: A
authors.lastname: Mitsueda
authors.forename: Yuri
authors.initials: Y
authors.lastname: Shimatani
authors.forename: Masahiro
authors.initials: M
authors.lastname: Ito
authors.forename: Takashi
authors.initials: T
authors.lastname: Ohgita
authors.forename: Asako
authors.initials: A
authors.lastname: Yamada
authors.forename: Susumu
authors.initials: S
authors.lastname: Hama
authors.forename: Astrid
authors.initials: A
authors.lastname: Gräslund
authors.forename: Staffan
authors.initials: S
authors.lastname: Lindberg
authors.forename: Ülo
authors.initials: Ü
authors.lastname: Langel
authors.forename: Hideyoshi
authors.initials: H
authors.lastname: Harashima
authors.forename: Ikuhiko
authors.initials: I
authors.lastname: Nakase
authors.forename: Shiroh
authors.initials: S
authors.lastname: Futaki
authors.forename: Kentaro
authors.initials: K
authors.lastname: Kogure
mesh_terms: D057846:Cell-Penetrating Peptides / Q000737:chemistry; D011992:Endosomes; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry; D034741:RNA, Small Interfering; D014162:Transfection
chemical_list: D057846:Cell-Penetrating Peptides; D034741:RNA, Small Interfering
keywords: cell-penetrating peptide; decondensation; siRNA delivery