2005 Fiscal Year Final Research Report Summary
Analysis of genes related to the ectopic bone formation induced by mechanical stress and search for drugs targeting the genes.
| Project/Area Number |
16390063
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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 |
General pharmacology
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| Research Institution | Hirosaki University |
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Principal Investigator |
FURUKAWA Ken-Ichi Hirosaki University School of Medicine, Department of Pharmacology, Associate Professor, 医学部, 助教授 (20165468)
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| Co-Investigator(Kenkyū-buntansha) |
INOUE Ituro The University of Tokyo, Division of genetic diagnosis, The Institute of Medical Science, Associate Professor, 医科学研究所, 客員助教授 (00192500)
OKADA Akihiro Hirosaki University Hospital, Orthopedic Surgery, Lecturer, 医学部附属病院, 講師 (90312488)
SEYA Kazuhiko Hirosaki University School of Medicine, Department of Pharmacology, Assistant, 医学部, 助手 (40281919)
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| Project Period (FY) |
2004 – 2005
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| Keywords | ectopic bone formation / mechanical stress / microarray / gene expression / siRNA / Cbfa1 / nucleotide receptor / endochondral ossification |
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| Research Abstract |
To clarify the cause of the ossification of posterior longitudinal ligament (OPLL), comprehensive analyses by cDNA microarray method were carried out using cultured human spinal ligament cells that had been subjected to uniaxial cyclic stretching (mechanical stress). Microarray analyses showed the up-regulation of many ossification-marker genes including alkaline phosphatase, osteocalcin, collagen type I and Cbfa1, by cyclic stretching in ligament cells from OPLL patients but not in normal ligament cells, suggesting that OPLL cells are higher sensitive to mechanical stress than normal ligament cells. The higher sensitivity to mechanical stress was also confirmed in ligament tissue from OPLL patients. These results suggest that mechanical stress participates in the progression of OPLL through changes in expressions of various genes related to bone metabolism.
Spectrum of P2 receptor subtype expression in OPLL cells was quite different from that in non-OPLL cells. Especially, P2Y1 expression in OPLL cells was much higher than that in non-OPLL cells and almost equal to that in SaOS2 (osteosarcoma cell line). ATP as a P2Y1 agonist increased the expressions of osteogenic markers such as ALP and osteopontin in OPLL cells but not in non-OPLL cells. These increases were almost diminished in the presence of P2Y1-specific antagonist. Furthermore, mechanical stress increased ATP release from OPLL cells remarkably. These results suggest that as one of extracellular factors influencing the process of ossification, a pathophysiological action of ATP via P2Y1 is important. They become a novel pharmacotherapeutic target for OPLL.
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