High gene delivery in tumor by intratumoral injection of tetraarginine-PEG lipid-coated protamine/DNA.
One obstacle to effective gene therapy lies in low transfection efficiency by non-viral vectors. To meet this challenge, we developed cell-penetrating peptide-based gene delivery vectors. A novel oligoarginine lipid ((Arg)n-B, n=4, 10) conjugated to 3,5-bis(dodecyloxy)benzamide (BDB) lipid with a poly(ethylene glycol) (PEG) spacer was synthesized. Oligoarginine lipid-coated vector was prepared by the addition of (Arg)n-B to DNA/protamine complex (PD) ((Arg)n-B-PD). Transfection efficiency of (Arg)n-B-PD was compared with that of (Arg)n-B/DNA complex ((Arg)n-B/D) for in vitro and in xenograft tumor of human cervical carcinoma HeLa by intratumoral injection. Transfection efficiency in tumors and in vitro greatly depended on the charge ratios of (Arg)n-B to DNA and the length of Arg residues. In vitro, positively charged Arg10-B-PD showed the highest transfection efficiency. In contrast, in tumor transfection, negatively charged Arg4-B-PD showed the highest transfection efficiency, about 2-, 16- and 23-fold higher than PD alone, Arg10-B-PD and a commercial gene transfection reagent, respectively. This result suggests that negatively charged tetraarginine-conjugated-PEG lipid-coated PD is a promising gene delivery vector for intratumoral injection.
Institute of Medicinal Chemistry, Hoshi University, Ebara 2-4-41, Shinagawa-ku, Tokyo 142-8501, Japan.
Takashi
T
Fujita
Masahiko
M
Furuhata
Yoshiyuki
Y
Hattori
Hiroko
H
Kawakami
Kazunori
K
Toma
Yoshie
Y
Maitani
D000818:Animals; D001120:Arginine / Q000737:chemistry*; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D015552:Injections, Intralesional; D008055:Lipids / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D009842:Oligopeptides / Q000737:chemistry*; D010316:Particle Size; D011092:Polyethylene Glycols / Q000737:chemistry*; D011479:Protamines / Q000737:chemistry*; D014162:Transfection / Q000379:methods*; D023041:Xenograft Model Antitumor Assays
D008055:Lipids; D009842:Oligopeptides; D011479:Protamines; D011092:Polyethylene Glycols; D004247:DNA; D001120:Arginine