Supramolecular assemblies for the cytoplasmic delivery of antisense oligodeoxynucleotide: polyion complex (PIC) micelles based on poly(ethylene glycol)-SS-oligodeoxynucleotide conjugate.
A novel cytoplasmic delivery system of antisense oligodeoxynucleotide (asODN) was developed by assembling a PEG-asODN conjugate with disulfide linkage (smart linkage) (PEG-SS-asODN) into polyion complex (PIC) micelles through the complexation with branched poly(ethylenimine) (B-PEI). The PIC micelle thus prepared showed a significant antisense effect against luciferase gene expression in HuH-7 cells, far more efficient than nonmicelle systems (asODN and PEG-SS-asODN in free form) and PIC micelle encapsulating the conjugate without the disulfide linkage. Use of poly(l-lysine) (PLL) instead of the B-PEI for PIC micellization led to a substantial decrease in the antisense effect. These results indicate that the PIC micelles formulated from PEG-SS-asODN conjugate and B-PEI is successfully transported from the endosomal compartment into the cytoplasm by the buffering effect of the B-PEI, releasing hundreds of active asODN molecules via cleavage of the disulfide linkage into the cellular interior, responding to a high glutathione concentration in the cytoplasmic compartment. Furthermore, the type of smart linkage (glutathione-sensitive SS linkage vs pH-sensitive linkage) in the conjugates substantially affected the antisense effect of the PIC micelles, depending on the nature of the counter polycation (B-PEI vs PLL).
Oishi
Motoi
M
Tsukuba Research Center for Interdisciplinary Materials Science (TIMS), University of Tsukuba, 1-1-1 Ten-noudai, Tsukuba, Ibaraki 305-8573, Japan.
Hayama
Tetsuya
T
Akiyama
Yoshitsugu
Y
Takae
Seiji
S
Harada
Atsushi
A
Yamasaki
Yuichi
Y
Nagatsugi
Fumi
F
Sasaki
Shigeki
S
Nagasaki
Yukio
Y
Kataoka
Kazunori
K
D001672:Biocompatible Materials / Q000737:chemistry*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D002412:Cations; D045744:Cell Line, Tumor; D002845:Chromatography; D003593:Cytoplasm / Q000378:metabolism*; D004220:Disulfides / Q000737:chemistry; D017930:Genes, Reporter; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007477:Ions; D008156:Luciferases / Q000378:metabolism; D008823:Micelles; D008954:Models, Biological; D008956:Models, Chemical; D016376:Oligonucleotides, Antisense / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D011107:Polylysine / Q000737:chemistry
D001672:Biocompatible Materials; D002412:Cations; D004220:Disulfides; D007477:Ions; D008823:Micelles; D016376:Oligonucleotides, Antisense; D011107:Polylysine; D011092:Polyethylene Glycols; D008156:Luciferases