Development of high-throughput screening system for osteogenic drugs using a cell-based sensor

Hironori Hojo; Kazuyo Igawa; Shinsuke Ohba; Fumiko Yano; Keiji Nakajima; Yuske Komiyama; Toshiyuki Ikeda; Alexander C. Lichtler; Je Tae Woo; Takayuki Yonezawa; Tsuyoshi Takato; Ung il Chung

doi:10.1016/j.bbrc.2008.08.167

Development of high-throughput screening system for osteogenic drugs using a cell-based sensor

Hironori Hojo, Kazuyo Igawa, Shinsuke Ohba, Fumiko Yano, Keiji Nakajima, Yuske Komiyama, Toshiyuki Ikeda, Alexander C. Lichtler, Je Tae Woo, Takayuki Yonezawa, Tsuyoshi Takato, Ung il Chung

Research output: Contribution to journal › Article › peer-review

20 Citations (Scopus)

Abstract

To effectively treat osteoporosis and other bone-loss disorders, small compounds that potently induce bone formation are needed. The present study initially attempted to establish a monitoring system that could detect osteogenic differentiation easily, precisely, and noninvasively. For this purpose, we established pre-osteoblastic MC3T3E1 cells stably transfected with the GFP reporter gene driven by a 2.3 kb fragment of rat type I collagen promoter (Col1a1GFP-MC3T3E1). Among these cells, we selected a clone that fluoresced upon osteogenic stimulation by BMP2. The GFP fluorescence intensity corresponded well to the intensity of alkaline phosphatase (ALP) staining and to the level of osteocalcin (Oc) mRNA. Using this system, we screened natural and synthetic compound libraries and thus identified an isoflavone derivative, glabrisoflavone (GI). GI induced ALP staining and Oc mRNA in a dose-dependent manner. The Col1a1GFP-MC3T3E1 system may be useful for identifying novel osteogenic drugs.

Original language	English
Pages (from-to)	375-379
Number of pages	5
Journal	Biochemical and Biophysical Research Communications
Volume	376
Issue number	2
DOIs	https://doi.org/10.1016/j.bbrc.2008.08.167
Publication status	Published - Nov 14 2008
Externally published	Yes

Keywords

Col1a1GFP-MC3T3E1
Osteoblast
Screening system
Small compounds

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.bbrc.2008.08.167

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@article{ef0c2203ecaf42cda54257daf2d74ba6,

title = "Development of high-throughput screening system for osteogenic drugs using a cell-based sensor",

abstract = "To effectively treat osteoporosis and other bone-loss disorders, small compounds that potently induce bone formation are needed. The present study initially attempted to establish a monitoring system that could detect osteogenic differentiation easily, precisely, and noninvasively. For this purpose, we established pre-osteoblastic MC3T3E1 cells stably transfected with the GFP reporter gene driven by a 2.3 kb fragment of rat type I collagen promoter (Col1a1GFP-MC3T3E1). Among these cells, we selected a clone that fluoresced upon osteogenic stimulation by BMP2. The GFP fluorescence intensity corresponded well to the intensity of alkaline phosphatase (ALP) staining and to the level of osteocalcin (Oc) mRNA. Using this system, we screened natural and synthetic compound libraries and thus identified an isoflavone derivative, glabrisoflavone (GI). GI induced ALP staining and Oc mRNA in a dose-dependent manner. The Col1a1GFP-MC3T3E1 system may be useful for identifying novel osteogenic drugs.",

keywords = "Col1a1GFP-MC3T3E1, Osteoblast, Screening system, Small compounds",

author = "Hironori Hojo and Kazuyo Igawa and Shinsuke Ohba and Fumiko Yano and Keiji Nakajima and Yuske Komiyama and Toshiyuki Ikeda and Lichtler, {Alexander C.} and Woo, {Je Tae} and Takayuki Yonezawa and Tsuyoshi Takato and Chung, {Ung il}",

note = "Funding Information: We thank D.W. Rowe, K. Miyazono, A. Hecht, G. Karsenty, and T. Katagiri for their kind provision of experimental materials. We also appreciate the helpful discussions with members of the Chung lab, University of Tokyo; K. Morii, Y. Hasegawa, M. Zenibayashi, K. Date, and K. Suzuki for providing technical assistance; and Astellas Pharma Inc. for providing rhBMP2. This work was supported by Grants-in-Aid for Scientific Research from the Japanese Ministry of Education, Culture, Sports, Science, and Technology (#17390530 and #20390509).",

year = "2008",

month = nov,

day = "14",

doi = "10.1016/j.bbrc.2008.08.167",

language = "English",

volume = "376",

pages = "375--379",

journal = "Biochemical and Biophysical Research Communications",

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T1 - Development of high-throughput screening system for osteogenic drugs using a cell-based sensor

AU - Hojo, Hironori

AU - Igawa, Kazuyo

AU - Ohba, Shinsuke

AU - Yano, Fumiko

AU - Nakajima, Keiji

AU - Komiyama, Yuske

AU - Ikeda, Toshiyuki

AU - Lichtler, Alexander C.

AU - Woo, Je Tae

AU - Yonezawa, Takayuki

AU - Takato, Tsuyoshi

AU - Chung, Ung il

N1 - Funding Information: We thank D.W. Rowe, K. Miyazono, A. Hecht, G. Karsenty, and T. Katagiri for their kind provision of experimental materials. We also appreciate the helpful discussions with members of the Chung lab, University of Tokyo; K. Morii, Y. Hasegawa, M. Zenibayashi, K. Date, and K. Suzuki for providing technical assistance; and Astellas Pharma Inc. for providing rhBMP2. This work was supported by Grants-in-Aid for Scientific Research from the Japanese Ministry of Education, Culture, Sports, Science, and Technology (#17390530 and #20390509).

PY - 2008/11/14

Y1 - 2008/11/14

N2 - To effectively treat osteoporosis and other bone-loss disorders, small compounds that potently induce bone formation are needed. The present study initially attempted to establish a monitoring system that could detect osteogenic differentiation easily, precisely, and noninvasively. For this purpose, we established pre-osteoblastic MC3T3E1 cells stably transfected with the GFP reporter gene driven by a 2.3 kb fragment of rat type I collagen promoter (Col1a1GFP-MC3T3E1). Among these cells, we selected a clone that fluoresced upon osteogenic stimulation by BMP2. The GFP fluorescence intensity corresponded well to the intensity of alkaline phosphatase (ALP) staining and to the level of osteocalcin (Oc) mRNA. Using this system, we screened natural and synthetic compound libraries and thus identified an isoflavone derivative, glabrisoflavone (GI). GI induced ALP staining and Oc mRNA in a dose-dependent manner. The Col1a1GFP-MC3T3E1 system may be useful for identifying novel osteogenic drugs.

AB - To effectively treat osteoporosis and other bone-loss disorders, small compounds that potently induce bone formation are needed. The present study initially attempted to establish a monitoring system that could detect osteogenic differentiation easily, precisely, and noninvasively. For this purpose, we established pre-osteoblastic MC3T3E1 cells stably transfected with the GFP reporter gene driven by a 2.3 kb fragment of rat type I collagen promoter (Col1a1GFP-MC3T3E1). Among these cells, we selected a clone that fluoresced upon osteogenic stimulation by BMP2. The GFP fluorescence intensity corresponded well to the intensity of alkaline phosphatase (ALP) staining and to the level of osteocalcin (Oc) mRNA. Using this system, we screened natural and synthetic compound libraries and thus identified an isoflavone derivative, glabrisoflavone (GI). GI induced ALP staining and Oc mRNA in a dose-dependent manner. The Col1a1GFP-MC3T3E1 system may be useful for identifying novel osteogenic drugs.

KW - Col1a1GFP-MC3T3E1

KW - Osteoblast

KW - Screening system

KW - Small compounds

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ER -

Development of high-throughput screening system for osteogenic drugs using a cell-based sensor

Abstract

Keywords

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