title: Particle tracking analysis for the intracellular trafficking of nanoparticles modified with African swine fever virus protein p54-derived peptide.
abstract: Previous studies showed that the cytoplasmic transport of nanoparticles to the nucleus is driven by a vesicular sorting system. Artificial approaches for targeting a microtubule-associating motor complex is also a challenge. We describe herein the development of a liposomal nanoparticle, the surface of which is modified with stearylated octa-arginine (STR-R8), and a dynein light chain (LC8)-associated peptide derived from an African swine fever virus protein p54 (p54(149-161)) with polyethyleneglycol (PEG) as a spacer (p54(149-161)-PEG/R8-liposomal nanoparticles (LNPs)). The p54(149-161)-PEG/R8-LNPs preferentially gain access to the nucleus, resulting in a one- to two-order of magnitude higher transfection activity in comparison with p54(149-161)-free nanoparticles (PEG/R8-LNPs). Further studies of particle tracking in HeLa cells stably expressing green fluorescent protein (GFP)-tagged tubulin (GFP/Tub-HeLa) indicate that p54(149-161) stimulated the transport of nanoparticles along fibrous tubulin structures. Moreover, a part of the p54(149-161)-PEG/R8-LNPs appeared to undergo quasi-straight transport without sharing the tracks corresponding to PKH67, the plasma membrane of which had been prestained with a marker just before transfection, while corresponding movement was never observed in the case of PEG/R8-LNPs. These findings suggest that a portion of the p54(149-161)-modified nanoparticles can use microtubule-dependent transport without the need for an assist by a vesicular sorting system.
authors.affiliation: Laboratory for Molecular Design of Pharmaceutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Akita
authors.forename: Kaoru
authors.initials: K
authors.lastname: Enoto
authors.forename: Hiroki
authors.initials: H
authors.lastname: Tanaka
authors.forename: Hideyoshi
authors.initials: H
authors.lastname: Harashima
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism; D018628:Gene Dosage; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008081:Liposomes; D008870:Microtubules / Q000235:genetics / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry*; D061848:Optical Imaging; D009930:Organic Chemicals / Q000378:metabolism; D010316:Particle Size; D010455:Peptides / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry; D021381:Protein Transport; D014162:Transfection; D015678:Viral Structural Proteins / Q000737:chemistry* / Q000235:genetics*
chemical_list: D008081:Liposomes; D009930:Organic Chemicals; C451241:PKH67; D010455:Peptides; D015678:Viral Structural Proteins; C093921:p54 protein, African swine fever virus; D049452:Green Fluorescent Proteins; D011092:Polyethylene Glycols