A fragment of the hypophosphatemic factor, MEPE, requires inducible cyclooxygenase-2 to exert potent anabolic effects on normal human marrow osteoblast precursors

D. E. Nagel, Sundeep Khosla, A. Sanyal, D. M. Rosen, Y. Kumagai, B. Lawrence Riggs

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

MEPE, 56.6 kDa protein isolated from tumors associated with hypophosphatemic osteomalacia, increases renal phosphate excretion and is expressed in normal human bone cells. AC-100, a central 23-amino acid fragment of MEPE, contains motifs that are important in regulating cellular activities in the bone microenvironment. Thus, we assessed in vitro effects of AC-100 on multipotential normal human marrow stromal (hMS) cells that have the capacity to differentiate into mature osteoblasts. Proliferation was quantified by [H 3]thymidine uptake and cell counting and differentiation by the levels of mRNA for the α2-chain of type I procollagen (COL1A2), alkaline phosphatase (AP), and osteocalcin (OC) measured using real time reverse transcriptase PCR (RT-PCR) and by the formation of mineralized nodules. AC-100 increased proliferation by 257 ± 89% (P < 0.005), increased gene expression of COL1A2 by 339 ± 85% (P < 0.005), AP by 1,437 ± 40% (P < 0.001), and OC by 1,962 ± 337% (P < 0.001). In addition, it increased mineralized nodule formation by 81 ± 14% (P < 0.001) in a dose- and time-dependent fashion. In equimolar dosages, the parent compound, MEPE, had the full activity of the AC-100 fragment. AC-100 elicited a comparable response to both IGF-1 and BMP-2 with respect to proliferation and differentiation of hMS cells. Using gene expression microarray analysis, we demonstrated that AC-100 increased (by ∼3-fold) the mRNA for cyclooxgenase-2 (COX-2), an inducible enzyme required for prostaglandin synthesis. Moreover, NS-398, a specific inhibitor of COX-2 action completely blocked AC-100-induced increases in proliferation and differentiation. Thus, AC-100 has potent anabolic activity on osteoblast precursor cells in vitro and these effects require the induction of COX-2.

Original languageEnglish (US)
Pages (from-to)1107-1114
Number of pages8
JournalJournal of Cellular Biochemistry
Volume93
Issue number6
DOIs
StatePublished - 2004

Fingerprint

Anabolic Agents
Osteocalcin
Osteoblasts
Cyclooxygenase 2
Gene expression
Alkaline Phosphatase
Bone
Bone Marrow
Stromal Cells
Messenger RNA
RNA-Directed DNA Polymerase
Microarrays
Collagen Type I
Insulin-Like Growth Factor I
Thymidine
Prostaglandins
Gene Expression
Bone and Bones
Tumors
Osteomalacia

Keywords

  • Anabolic agent
  • BMP-2
  • Bone formation
  • COX-2

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

A fragment of the hypophosphatemic factor, MEPE, requires inducible cyclooxygenase-2 to exert potent anabolic effects on normal human marrow osteoblast precursors. / Nagel, D. E.; Khosla, Sundeep; Sanyal, A.; Rosen, D. M.; Kumagai, Y.; Riggs, B. Lawrence.

In: Journal of Cellular Biochemistry, Vol. 93, No. 6, 2004, p. 1107-1114.

Research output: Contribution to journalArticle

@article{d1f0eb20d5a44885a838d8735823ce09,
title = "A fragment of the hypophosphatemic factor, MEPE, requires inducible cyclooxygenase-2 to exert potent anabolic effects on normal human marrow osteoblast precursors",
abstract = "MEPE, 56.6 kDa protein isolated from tumors associated with hypophosphatemic osteomalacia, increases renal phosphate excretion and is expressed in normal human bone cells. AC-100, a central 23-amino acid fragment of MEPE, contains motifs that are important in regulating cellular activities in the bone microenvironment. Thus, we assessed in vitro effects of AC-100 on multipotential normal human marrow stromal (hMS) cells that have the capacity to differentiate into mature osteoblasts. Proliferation was quantified by [H 3]thymidine uptake and cell counting and differentiation by the levels of mRNA for the α2-chain of type I procollagen (COL1A2), alkaline phosphatase (AP), and osteocalcin (OC) measured using real time reverse transcriptase PCR (RT-PCR) and by the formation of mineralized nodules. AC-100 increased proliferation by 257 ± 89{\%} (P < 0.005), increased gene expression of COL1A2 by 339 ± 85{\%} (P < 0.005), AP by 1,437 ± 40{\%} (P < 0.001), and OC by 1,962 ± 337{\%} (P < 0.001). In addition, it increased mineralized nodule formation by 81 ± 14{\%} (P < 0.001) in a dose- and time-dependent fashion. In equimolar dosages, the parent compound, MEPE, had the full activity of the AC-100 fragment. AC-100 elicited a comparable response to both IGF-1 and BMP-2 with respect to proliferation and differentiation of hMS cells. Using gene expression microarray analysis, we demonstrated that AC-100 increased (by ∼3-fold) the mRNA for cyclooxgenase-2 (COX-2), an inducible enzyme required for prostaglandin synthesis. Moreover, NS-398, a specific inhibitor of COX-2 action completely blocked AC-100-induced increases in proliferation and differentiation. Thus, AC-100 has potent anabolic activity on osteoblast precursor cells in vitro and these effects require the induction of COX-2.",
keywords = "Anabolic agent, BMP-2, Bone formation, COX-2",
author = "Nagel, {D. E.} and Sundeep Khosla and A. Sanyal and Rosen, {D. M.} and Y. Kumagai and Riggs, {B. Lawrence}",
year = "2004",
doi = "10.1002/jcb.20249",
language = "English (US)",
volume = "93",
pages = "1107--1114",
journal = "Journal of Cellular Biochemistry",
issn = "0730-2312",
publisher = "Wiley-Liss Inc.",
number = "6",

}

TY - JOUR

T1 - A fragment of the hypophosphatemic factor, MEPE, requires inducible cyclooxygenase-2 to exert potent anabolic effects on normal human marrow osteoblast precursors

AU - Nagel, D. E.

AU - Khosla, Sundeep

AU - Sanyal, A.

AU - Rosen, D. M.

AU - Kumagai, Y.

AU - Riggs, B. Lawrence

PY - 2004

Y1 - 2004

N2 - MEPE, 56.6 kDa protein isolated from tumors associated with hypophosphatemic osteomalacia, increases renal phosphate excretion and is expressed in normal human bone cells. AC-100, a central 23-amino acid fragment of MEPE, contains motifs that are important in regulating cellular activities in the bone microenvironment. Thus, we assessed in vitro effects of AC-100 on multipotential normal human marrow stromal (hMS) cells that have the capacity to differentiate into mature osteoblasts. Proliferation was quantified by [H 3]thymidine uptake and cell counting and differentiation by the levels of mRNA for the α2-chain of type I procollagen (COL1A2), alkaline phosphatase (AP), and osteocalcin (OC) measured using real time reverse transcriptase PCR (RT-PCR) and by the formation of mineralized nodules. AC-100 increased proliferation by 257 ± 89% (P < 0.005), increased gene expression of COL1A2 by 339 ± 85% (P < 0.005), AP by 1,437 ± 40% (P < 0.001), and OC by 1,962 ± 337% (P < 0.001). In addition, it increased mineralized nodule formation by 81 ± 14% (P < 0.001) in a dose- and time-dependent fashion. In equimolar dosages, the parent compound, MEPE, had the full activity of the AC-100 fragment. AC-100 elicited a comparable response to both IGF-1 and BMP-2 with respect to proliferation and differentiation of hMS cells. Using gene expression microarray analysis, we demonstrated that AC-100 increased (by ∼3-fold) the mRNA for cyclooxgenase-2 (COX-2), an inducible enzyme required for prostaglandin synthesis. Moreover, NS-398, a specific inhibitor of COX-2 action completely blocked AC-100-induced increases in proliferation and differentiation. Thus, AC-100 has potent anabolic activity on osteoblast precursor cells in vitro and these effects require the induction of COX-2.

AB - MEPE, 56.6 kDa protein isolated from tumors associated with hypophosphatemic osteomalacia, increases renal phosphate excretion and is expressed in normal human bone cells. AC-100, a central 23-amino acid fragment of MEPE, contains motifs that are important in regulating cellular activities in the bone microenvironment. Thus, we assessed in vitro effects of AC-100 on multipotential normal human marrow stromal (hMS) cells that have the capacity to differentiate into mature osteoblasts. Proliferation was quantified by [H 3]thymidine uptake and cell counting and differentiation by the levels of mRNA for the α2-chain of type I procollagen (COL1A2), alkaline phosphatase (AP), and osteocalcin (OC) measured using real time reverse transcriptase PCR (RT-PCR) and by the formation of mineralized nodules. AC-100 increased proliferation by 257 ± 89% (P < 0.005), increased gene expression of COL1A2 by 339 ± 85% (P < 0.005), AP by 1,437 ± 40% (P < 0.001), and OC by 1,962 ± 337% (P < 0.001). In addition, it increased mineralized nodule formation by 81 ± 14% (P < 0.001) in a dose- and time-dependent fashion. In equimolar dosages, the parent compound, MEPE, had the full activity of the AC-100 fragment. AC-100 elicited a comparable response to both IGF-1 and BMP-2 with respect to proliferation and differentiation of hMS cells. Using gene expression microarray analysis, we demonstrated that AC-100 increased (by ∼3-fold) the mRNA for cyclooxgenase-2 (COX-2), an inducible enzyme required for prostaglandin synthesis. Moreover, NS-398, a specific inhibitor of COX-2 action completely blocked AC-100-induced increases in proliferation and differentiation. Thus, AC-100 has potent anabolic activity on osteoblast precursor cells in vitro and these effects require the induction of COX-2.

KW - Anabolic agent

KW - BMP-2

KW - Bone formation

KW - COX-2

UR - http://www.scopus.com/inward/record.url?scp=21644453949&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=21644453949&partnerID=8YFLogxK

U2 - 10.1002/jcb.20249

DO - 10.1002/jcb.20249

M3 - Article

C2 - 15449321

AN - SCOPUS:21644453949

VL - 93

SP - 1107

EP - 1114

JO - Journal of Cellular Biochemistry

JF - Journal of Cellular Biochemistry

SN - 0730-2312

IS - 6

ER -