| Project/Area Number |
14370400
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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 | 一般 |
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| Research Field |
Thoracic surgery
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| Research Institution | Tohoku University |
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Principal Investigator |
TABAYASHI Koichi Tohoku University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (90142942)
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| Co-Investigator(Kenkyū-buntansha) |
MIYAZAKI Jun-ichi Osaka University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (10200156)
IGUCHI Atsushi Tohoku University, Graduate School of Medicine, Associate Professor, 大学院・医学系研究科, 助教授 (90222851)
NITTA Yoshio Tohoku University, Hospital, Research Associate, 病院・助手 (80375005)
遠藤 雅人 東北大学, 大学病院, 講師 (90282128)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥4,700,000 (Direct Cost: ¥4,700,000)
Fiscal Year 2004: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2002: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | spinal cord protection / thoracic aneurysm / gene therapy / paraplegia / ORP150 / CAG promoter / adenoviral vector / lipofection / eNOS / 遺伝子導入 / 虚血再潅流障害 / 脊髄虚血 / 精髄保護 / spinal cord injury / lipofection / naked DNA / denovival vector / spinal code injury / adenoviral vector |
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| Research Abstract |
Spinal cord injury after successful aortic surgical operations is an unacceptable complication for patients, and new spinal cord protection method is required. Oxygen-regulated protein 150kD (ORP150) is a novel endoplasmic-reticulum-associated chaperone induced by hypoxia/ischemia. And it was reported that cultured neurons overexpressing ORP150 were resistant to hypoxemic stress.
We hypothesized that overexpression of Oxygen-regulated protein 150kD(ORP150) in spinal cord would reduced motor neuron death after ischemia. ORP150 cDNA was obtained from RT-PCR products of Rat brain, and was inserted into expressing plasmid driven by CAG promoter. Expression of ORP150 was confirmed by using Western blotting method. Though in vivo lipofection of ORP150 or LacZ expression plasmid were tried to rabbit spinal cord, transduced gene were not detected at all. Neurological score after lipofecton reduced as compared to sham control. ORP150 expressing adenovirus vector was constructed by using circular form of the adenoviral genome cloned in a cosmid and the Cre-loxP recombination system. However, enough amount of adenoviral particles could not be obtained. As CAG promoter, an actin-based hybrid promoter, has very strong activities in most cells, excess expression of ORP150 was considered to have toxicity against 293 cells. Though two different gene delivery methods were planed for spinal protection, results were limited. These findings suggested that excess expression of ORP150 potentially also had toxicity against nerve cells, and to manage the expression levels of ORP150 is considered to be important when we apply this method to aortic surgery.
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