Expression Cloning of Joint Regenerative Genes
| Project/Area Number |
17591549
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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 |
Orthopaedic surgery
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| Research Institution | Saitama Medical University (2006)
The University of Tokyo (2005) |
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Principal Investigator |
MATSUBARA Takehiro Saitama Medical University, Faculty of Medicine, Associate, 医学部, 助手 (40361498)
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| Co-Investigator(Kenkyū-buntansha) |
IKEDA Toshiyuki Tokyo Univ, Faculty of Medicine, Visiting Research Associate, 医学部附属病院, 客員教員 (80322759)
ITAKA Keishi Tokyo Univ of Tokyo, Graduate School of Medicine, Project Lecture, 大学院医学系研究科, 特任講師 (60292926)
TEI Yuichi Tokyo Univ of Tokyo, Graduate School of Medicine, Professor, 大学院医学系研究科, 教授 (30345053)
KAWAGUCHI Hiroshi Tokyo Univ of Tokyo, Faculty of Medicine, Associate Professor (40282660)
NAKAMURA Kozo Tokyo Univ of Tokyo, Faculty of Medicine, Professor (60126133)
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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 |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2006: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2005: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | Cartilage Metabolism / Joint Development / Transcription Factor / 転写制御 / WNT9A / GDF5 / HMG / ATX |
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| Research Abstract |
To understand molecular mechanism of joint development, transcriptional regulation of three joint marker genes, GDF5, WNT9A and autototaxin were analyzed by luciferase-reporter assay. We identified a highly conserved 42 by core enhancer in the proximal promoter of WNT9A and named it JSE (joint specific enhancer). To identify common regulatory signal of cartilage development including articular cartilage, we searched the promoter region of SOX6, a key transcription factor for chondrogenesis. By combining 5'RACE, comparative genomics and luciferase-reporter assay, we identified the human embryonic SOX6 promoter and a 46 by core enhancer highly conserved among species and named it CES6 (core enhancer of SOX6). To identify regulatory transcription factors of cartilage and joint development, we performed southwestern screening by combining phage display libraries derived from cartilage tissues and mouse embryos and bait DNA sequences of 4x JSE and 4x CES6, and identified two transcription f
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actors HMGB2 and p63 that transactivated both WNT9A and SOX6 promoter. Further analysis revealed that p63 directly bound proximal promoters of GDF5, SOX6 and WNT9A and activated them. The binding sequences of p63 in these three genes were 1-3 by different from the known p53 family binding motif "RRRCWWGYYY". We speculated that p63 would recognize looser motifs in wide varieties of genes and regulate successive steps of chondrogenesis including joint formation. On the other hand, retroviral overexpression of p63 in mouse teratocarcinoma derived chondrogenic cell line ATDC5 induced Sox6 and Col2a1 mRNA strongly and rarely induced Gdf5 and Wnt9a. Joint specific transcription factors should be needed for optimal induction of joint marker genes. We established the technique to transfer adenoviral vectors in ex-vivo organn cultured developmental limbs derived from mouse embryos. WNT9A and p63 overexpressions by adenoviral vectors in developmental mouse limbs failed to induce joint space formations, however. To establish a more specific and physiological ex-vivo joint formation system, the information on more joint specific transcription factors for joint formations will be needed. We believe that applying results of this study for further investigations will reveal sufficient signals for joint formations. Less
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Report
(3 results)
Research Products
(11 results)
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[Journal Article] SOX2006
Author(s)
Saito T et al.
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Journal Title
Treatment of Osteoporosis 5 (1)
Pages: 70-73
Description
「研究成果報告書概要(欧文)」より
Related Report
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