title: Development of lipid particles targeted via sugar-lipid conjugates as novel nuclear gene delivery system.
abstract: Efficient nuclear gene delivery is essential for successful gene therapy. This study developed a novel system that mimics the mechanism of nuclear entry of adenovirus (Ad) by means of a Multifunctional Envelope-type Nano Device (MEND). In this system, plasmid DNA (pDNA) was condensed with polycation, followed by encapsulation in a lipid membrane. To target MEND to the nuclear pore complex (NPC), sugar served as a NPC-mediated nuclear targeting device was modified on the surface of the lipid envelope. This was accomplished via synthesis of a sugar-cholesterol conjugate. After binding of the MEND to the NPC, the pDNA core was transferred into the nucleus in conjunction with a breakdown of the lipid envelope. Sugar-modified MEND showed higher transfection efficiency compared with unmodified MEND, in non-dividing and dividing cells. Confocal microscopy confirmed that nuclear transfer of pDNA was improved by sugar modification of MEND. Furthermore, destabilization of the lipid envelope significantly enhanced transfection activity: therefore, nuclear-delivery efficiency was closely related to lipid envelope stability. Moreover, quantitative evaluation of cellular uptake and nuclear transfer processes by real-time PCR confirmed that the surface sugars affected nuclear transfer, but not cellular uptake. In summary, a novel system for the nuclear delivery of pDNA was successfully developed by using a sugar-modified MEND and by optimizing the lipid envelope stability.
authors.affiliation: Faculty of Pharmaceutical Sciences, Hokkaido University, Kita 12 Nishi 6, Kita-Ku, Sapporo, Hokkaido 060-0812, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Masuda
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Akita
authors.forename: Takashi
authors.initials: T
authors.lastname: Nishio
authors.forename: Kenichi
authors.initials: K
authors.lastname: Niikura
authors.forename: Kentaro
authors.initials: K
authors.lastname: Kogure
authors.forename: Kuniharu
authors.initials: K
authors.lastname: Ijiro
authors.forename: Hideyoshi
authors.initials: H
authors.lastname: Harashima
mesh_terms: D001483:Base Sequence; D002241:Carbohydrates / Q000737:chemistry*; D017931:DNA Primers; D018014:Gene Transfer Techniques; D008055:Lipids / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D018613:Microscopy, Confocal; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D002241:Carbohydrates; D017931:DNA Primers; D008055:Lipids