2007 Fiscal Year Final Research Report Summary
Generation of gene therapeutic method for diabetic neuropathy by AAV vector
| Project/Area Number |
18590934
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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 |
Neurology
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| Research Institution | Shiga University of Medical Science |
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Principal Investigator |
YASUDA Hitoshi Shiga University of Medical Science, Faculty of Nursing, Professor (80135467)
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| Co-Investigator(Kenkyū-buntansha) |
KOJIMA Hideto Shiga University of Medical Science, Faculty of Medicine, Associate Professor (00225434)
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| Project Period (FY) |
2006 – 2007
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| Keywords | diabetic neuropathy / dorsal root ganglion / phage vector / RT-PCR / cDNA / erythropoietin / neuronal growth factor |
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| Research Abstract |
Phage display is a powerful technology to identify peptide sequence motifs that target a particular tissue or cell type in the body. Coupling such peptides to drugs and genes would enable their targeted delivery to specific cells and tissues in vitro and in vivo. In this research, we isolated peptides that home to mouse dorsal root ganglion (DRG) from a phage library expressing random7-mer peptides fused to a minor coat protein (pIII) of the M13phage. An in vitro biopanning procedure yielded 113phage plaques after 5 cycles of enrichment by incubation with isolated DRG neurons and two cycles of subtraction by exposure to irrelevant cell lines. Analyses of the sequences of this collection identified three peptide clones that occurred repeatedly during the biopanning procedure. Phage-antibody staining revealed that the three peptides bound to DRG neurons of different sizes. To determine if the peptides would recognize neuronal cells in vivo, we injected individual GST-peptide-fusion proteins into the subarachnoid space of mice and observed the appearance of immunoreactive GST in the cytosol of DRG neurons with a similar size distribution as that observed in vitro, indicating that the GST-peptide-fusion proteins were recognized and taken up by different DRG neurons in vivo. The identification of homing peptide sequences provides a powerful tool for future studies on DRG neuronal function in vitro and in vivo, and opens up the possibility of neuron-specific drug and gene delivery in the treatment of diseases affecting DRG neurons.
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Research Products
(2 results)
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[Journal Article] Isolation of specific peptides that home to dorsal root ganglion neurons in mice.2008
Author(s)
Oi, J., Terashima, T., Kojima, H.,Fujimia, M., Maeda, K., Arai, R., Chan. L., Yasud,. H., Kashiwagi, A., Kimura, H
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Journal Title
Neuroscience Letters 434-3
Pages: 266-272
Description
「研究成果報告書概要(欧文)」より
