| 학회 | 한국고분자학회 |
| 학술대회 | 2005년 가을 (10/13 ~ 10/14, 제주 ICC) |
| 권호 | 30권 2호 |
| 발표분야 | 고분자를 이용한 유전자 전달 |
| 제목 | Cytokine gene delivery using biomembrane or cationic polymers |
| 초록 | I) Gene delivery to progenitor cells using cationic polymers With the emerging role of hematopoietic stem cells as potential gene and cell therapy vehicles, there is an increasing need for safe and effective nonviral gene delivery systems. Gene transfer and transfection efficiency in human hematopoietic and cord blood CD34(+) cells could be enhanced by the use of low molecular weight polyethylenimine (PEI). PEIs of various molecular weights (800-750,000) were tested, and our results showed that the uptake of plasmid DNA by hematopoietic TF-1 cells depended on the molecular weights and the N/P ratios. Treatment with PEI 2K (m.w. 2000) at an N/P ratio of 80/1 was most effective, increasing the uptake of plasmid DNA in TF-1 cells by 23-fold relative to Lipofectamine 2000. PEI 2K-enhanced transfection was similarly observed in hematopoietic K562, murine Sca-1(+), and human cord blood CD34(+) cells. Notably, in human CD34(+) cells, a model gene transferred with PEI 2K showed 21,043- and 513-fold higher mRNA expression levels relative to the same construct transfected without PEI or with PEI 25 K, respectively. Moreover, PEI 2K-treated TF-1 and human CD34(+) cells retained good viability. Collectively, these results indicate that PEI 2K at the optimal N/P ratio might be used to safely enhance gene delivery and transfection of hematopoietic and human CD34(+) stem cells. II) Gene delivery using erythrocyte ghosts Red blood cell-derived erythrocyte ghosts (EG) were studied as a biocompatible nonviral delivery system for extended circulation and prolonged expression of plasmid DNA in the blood. Murine interleukin-2-expressing plasmid DNA was efficiently loaded to EG by electroporation in hypotonic condition. The presence of plasmid DNA in EG was confirmed by fluorescence-labeled plasmid DNA. At 21 min after intravenous administration into mice, the level of plasmid DNA in the blood was 92 000-fold higher following EG-mediated delivery as compared to the injection of naked form. EG-mediated gene delivery revealed higher and more prolonged mRNA expression levels of plasmid DNA in the blood until 9 days after the single intravenous injection. Moreover, plasmid DNA-loaded EG showed gene expression targeted to the blood cells. At 3 days post-dose, substantial expression levels of plasmid DNA delivered in EG were observed only in the blood and not in the other organs. Of the blood cells, the subpopulation containing granulocytes showed higher expression of plasmid DNA than mononuclear cells. These results indicate the potential of EG as a safe, prolonged and blood-targeted delivery system of therapeutic genes. |
| 저자 | Hyang-Min Byun, Ji-Young Shin, Yu-Kyoung Oh |
| 소속 | 고려대 |
| 키워드 | |
