| Project/Area Number |
15590690
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | Juntendo University |
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Principal Investigator |
TAKEI Yoshiyuki Juntendo University, Dept.of Gastroenterology, Associate Professor, 医学部, 助教授 (10306954)
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| Co-Investigator(Kenkyū-buntansha) |
IKEJIMA Kenichi Juntendo University, Dept.of Gastroenterology, Assistant Professor, 医学部, 講師 (20317382)
ENOMOTO Nobuyuki Juntendo University, Dept.of Gastroenterology, Assistant Professor, 医学部, 助手 (20348973)
MARUYAMA Atsushi Kyushu University, Institute for Materials Chemistry and Engineering, Professor, 先導物質化学研究所, 教授 (40190566)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2005: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | sinusoidal endothelial cells / gene delivery / triplex DNA / 遺伝子デリバリ / 相分離性ナノ会合体 / デコイ法 / 3重鎖核酸法 / ヒアルロン酸グラフト化ポリLリジン櫛型共重合体 / リホザイムライブラリ / 再生 / ヒアルロン酸グラフト化ポリLリジン櫛形共重合体 / ステルス効果 |
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| Research Abstract |
Altered gene expression in liver sinusoidal endothelial cells (SECs) is associated with a variety of aspects of liver pathophysiology. It is thus possible to envision a new therapeutic strategy for treatment of intractable liver diseases and achievement of graft-specific immunotolerance through modulation of SEC functions by genetic engineering. The SEC possesses unique hyaluronan receptors that recognize and internalize hyaluronic acid (HA). This characteristic was used in the development of a system for targeting foreign DNA to SECs. A gene carrier system was prepared by coupling HA oligomers to poly-L-lysine (PLL) in a 1:1 weight ratio by reductive amination reaction. The resulting copolymer (PLL-g-HA) was mixed with various amounts of DNA in 154 mM NaCl. Inter-polyelectrolyte complex formation between PLL-g-HA and DNA exhibited minimal self-aggregation, explaining the highly soluble nature of the complex. Complex formation between PLL-g-HA and DNA was further assessed with a gel re
… More
tardation assay. The titration point representing the minimum proportion of PLL-g-HA required to retard the DNA completely occurred at a 1:1 copolymer (based on PLL) to DNA charge ratio. Following intravenous injection of ^<32>P-labeled pSV β-Gal plasmid complexed to PLL-g-HA in Wistar rats, >90% of the injected counts were shown to be taken up by the liver. Further, it was shown that the PLL-g-HA/DNA complex was distributed exclusively in the SECs. Seventy-two hours after injection of 90 μg of pSV β-Gal in a PLL-g-HA-complexed form, a large number of SECs expressing β-galactosidase were detected. Thus, the PLL-g-HA/DNA system permits targeted delivery of exogenous nucleotide agents selectively to the liver SECs. We have focused on the oligonucleotide-based techniques such as RNAi, decoy and triplex DNA for manipulation gene expression in the SECs. NF-kB decoy oligomer in combination with PLL-g-HA inhibited effectively and specifically TNFα-induced ICAM-1 expression. Moreover, the PLL-g-HA/DNA carrier system has been shown to stabilize triplex DNA. Thus, the gene carrier system may provide a new and more effective strategy for manipulation of SEC functions including immunoresponses. Less
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