| Project/Area Number |
17590136
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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 |
Medical pharmacy
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| Research Institution | Showa Pharmaceutical University |
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Principal Investigator |
WATANABE Yoshiteru Showa Pharmaceutical University, Professor, 薬学部, 教授 (70175131)
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| Co-Investigator(Kenkyū-buntansha) |
KONDOH Masuo Osaka University, Graduate School, Associate Professor, 大学院・薬学研究科, 准教授 (50309697)
FUJII Makiko Showa Pharmaceutical University, Associate Professor, 薬学部, 准教授 (50199296)
KOIZUMI Naoya Showa Pharmaceutical University, Research Associate, 薬学部, 助教 (80433845)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2005: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | placental barrier / drug permeation / RNA interference / gene delivery system / transgene expression / trophoblast cell / pharmacology / 遺伝子 / 薬物透過機構 |
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| Research Abstract |
To elucidate drug permeation mechanism by transporters in placental barrier, we positively studied the establishment of RNA interference (RNAi) technique using viral and nonviral vector systems. In a preliminary study, RNAi using fiber-modified Ad vectors, it was suggested that the combination of fiber-modified Ad vectors containing polylysine peptide and RNAi is an effective tool for study of biological and physiological mechanism such as adipogenesis in adipose. Ad vector-mediated RNAi for peroxisome proliferator-activated receptor (PPAR) γ should also be useful to clarify the biological role of the PPAR γ pathway in various tissues. This finding led us to investigate RNAi using Ad vectors in placental barrier functions. On the other hand, we studied transgene activity of Ad vector containing Arg-Gly-Asp (RGD) motif in the fiber protein (Ad-RGD vector) during differentiation of human cytotrophoblast BeWo cells into syncytiotrophoblast-like cells. Ad-RGD vector had no influence on dif
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ferentiation and had a 2.5 fold higher transduction activity than that of the conventional Ad vector. Thus, Ad-vRGD vector can be a powerful tool for gene transfer experiments in syncytiotrophoblast cells. Subsequently, we investigated characteristics of transcription-regulatory elements for gene expression from plasmid vectors in human trophoblast cell lines BeWo and JAR cells. It was found that insertion of intron elements enhanced transcriptional activities in the following order: intron A>hybrid β-globin-immunoglobin intron>no intron. We also found that gene expression level can be controlled by selecting the expression cassette. These results may be useful for further studies on construction of siRNA expression cassette by polymerase II promoter. For induction of transporters, we observed an induction of zinc transporters by forskolin in human trophoblast BeWo cells. A function of transporter in placenta during pregnancy, the mechanism of zinc release from the placenta into fetal circulation is not well understand. Treatment of BeWo cells with forskolin, a representative inducer of differentiation of BeWo cells cytotrophoblast into syncytiotrophoblast, resulted in morphological changes of BeWo cells and elevated Zn transporter 1, 2 and 4 mRNA levels. Several findings in this project should be useful for studies aimed at analysis of transporters in placenta using RNAi. We will continue further studies on RNAi of transporters by delivery of viral and nonviral vectors. Less
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