Gamma-polyglutamic acid-coated vectors for effective and safe gene therapy.
In the present study, we developed some novel gene delivery vectors, coated cationic complexes with gamma-polyglutamic acid (gamma-PGA) for effective and safe gene therapy. Cationic complexes were constructed with pDNA and cationic vectors, such as poly-L-arginine hydrochloride (PLA), poly-L-lysine hydrobromide (PLL), N-[1-(2, 3-dioleyloxy) propyl]-N, N, N-trimethylammonium chloride (DOTMA)-cholesterol (Chol) liposomes, and DOTMA-dioleylphosphatidylethanolamine (DOPE) liposomes. The cationic complexes showed high gene expression with strong cytotoxicity in melanoma B16-F10 cells. The cationic complexes were also strongly toxic to erythrocytes. On the other hand, the gamma-PGA was able to coat all cationic complexes and form stable nano-sized particles with negative charges. These gamma-PGA-coated complexes had high gene expression without cytotoxicity and toxicities to the erythrocytes. In in vivo transfection experiments, polyplexes showed high transfection efficiency over 10(5) RLU/g in the lung tissue after intravenous injection, although gamma-PGA-coated polyplexes showed a high value in the spleen. High transfection efficiency in lipoplexes and gamma-PGA-coated lipoplexes was observed in the spleen and lung. Thus, gamma-PGA-coated vectors are useful for clinical gene therapy.
Kurosaki
Tomoaki
T
Department of Hospital Pharmacy, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
Kitahara
Takashi
T
Kawakami
Shigeru
S
Higuchi
Yuriko
Y
Yamaguchi
Ayumi
A
Nakagawa
Hiroo
H
Kodama
Yukinobu
Y
Hamamoto
Tomoyuki
T
Hashida
Mitsuru
M
Sasaki
Hitoshi
H
D000371:Agglutination / Q000187:drug effects; D000818:Animals; D002412:Cations; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D004337:Drug Carriers / Q000737:chemistry* / Q000633:toxicity; D004912:Erythrocytes / Q000187:drug effects; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D006461:Hemolysis / Q000187:drug effects; D008081:Liposomes; D051379:Mice; D008856:Microscopy, Fluorescence; D009928:Organ Specificity; D010455:Peptides / Q000737:chemistry / Q000633:toxicity; D055668:Photochemical Processes; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D011099:Polyglutamic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000633:toxicity; D011107:Polylysine / Q000737:chemistry / Q000633:toxicity; D014162:Transfection
D002412:Cations; D004337:Drug Carriers; D008081:Liposomes; D010455:Peptides; C511775:poly(gamma-glutamic acid); D011107:Polylysine; C015462:polyarginine; D011099:Polyglutamic Acid; D004247:DNA