2002 Fiscal Year Final Research Report Summary
B型肝炎ウイルス表面抗原ナノ粒子を用いる生体内ピンポイント遺伝子導入法の開発
| Project/Area Number |
13558110
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | Osaka University |
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Principal Investigator |
TANIZAWA Katsuyuki Osaka University, Institute of Scientific and Industrial Research, Professor, 産業科学研究所, 教授 (20133134)
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| Project Period (FY) |
2001 – 2002
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| Keywords | genetherapy / drug delivery system / hepatitis B virus / yeast / liver / nanoparticles |
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| Research Abstract |
We previously succeeded overproduction of the HBV env L particles in yeast cells (up to 42% of the total soluble protein). In the present studies, the L particles have been purified, characterized, and examined for the applicability to the gene delivery system. By AFM observation and sedimentation equilibrium, about 110 molecules of L proteins were found to be assembled into a lipid vesicle to form a spherical particle (-500 nm in diameter). To examine the L particles as gene carriers, a mammalian expression plasmid for GFP (green fluorescence protein) was incorporated into L particles by electroporation. The L particles containing the plasmid were added to the culture medium of human hepatoma HepG2 cells. After two days, more than 90% of the HepG2 cells expressed GFP, while the control non-human liver cells did not. Then, the nude mice transplanted with human hepatoma HuH-7 cells and human colon cancer WiDr cells were injected intraperitoneaUy with the L particles containing the plasmid. Two weeks later, the fluorescence was observed specifically in the HuH-7 cells, but neither in the WiDr cells nor in the liver, spleen, kidney, and intestine of the mice. Because the L particle is an empty vesicle containing no viral DNA, it can be used as a safe and efficient vector for human liver-specific gene transfer. We are now evaluating the effectiveness of L particles as the novel drug delivery system, together with the genetically engineered L particles that can be applied for the pinpoint gene/drug delivery system to different tissues.
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[Publications] Yamada, T., Iwabuki, H., Kanno, T., Tanaka, H., Kawai, T., Fukuda, H.,Kondo, A., Seno, M., Tanizawa, K., and Kuroda, S.: "Physicochemical and Immunological Characterization of Hepatitis B Virus Envelope Particles Exclusively Consisting of the Entire L (Pre-S1+Pre-S2+S) Protein."Vaccine. 19. 3154-3163 (2001)
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「研究成果報告書概要(欧文)」より
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[Publications] Kanno, T., Yamada, T., Iwabuki, H., Tanaka, H., Kuroda, S., Tanizawa, K.,and Kawai, T.: "Size Distribution Measurement of Vesicles by Atomic Force Microscopy."Anal. Biochem.. 309. 196-199 (2002)
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「研究成果報告書概要(欧文)」より
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[Publications] Seno, M., Kuroda, S., Kondo, A., Tada, H., Tanizawa, K., and Ueda, M.: "Pinpoint Delivery System for Genes and Drugs with Bio-Nanoparticles."BIO INDUSTRY (in Japanese). No.4. 54-64 (2003)
Description
「研究成果報告書概要(欧文)」より
