Induction of Osteoblast Differentiation by Selective Activation of Kinase-Mediated Actions of the Estrogen Receptor

Abstract

Estrogens control gene transcription by cis or trans interactions of the estrogen receptor (ER) with target DNA or via the activation of cytoplasmic kinases. We report that selective activation of kinase-mediated actions of the ER with 4-estren-3α,17β-diol (estren) or an estradiol-dendrimer conjugate, each a synthetic compound that stimulates kinase-mediated ER actions 1,000 to 10,000 times more potently than direct DNA interactions, induced osteoblastic differentiation in established cell lines of uncommitted osteoblast precursors and primary cultures of osteoblast progenitors by stimulating Wnt and BMP-2 signaling in a kinase-dependent manner. In sharp contrast, 17β-estradiol (E₂) suppressed BMP-2-induced osteoblast progenitor commitment and differentiation. Consistent with the in vitro findings, estren, but not E₂, stimulated Wnt/β-catenin-mediated transcription in T-cell factor-lacZ transgenic mice. Moreover, E₂ stimulated BMP signaling in mice in which ERα lacks DNA binding activity and classical estrogen response element-mediated transcription (ERα^NERKI/−) but not in wild-type controls. This evidence reveals for the first time the existence of a large signalosome in which inputs from the ER, kinases, bone morphogenetic proteins, and Wnt signaling converge to induce differentiation of osteoblast precursors. ER can either induce it or repress it, depending on whether the activating ligand (and presumably the resulting conformation of the receptor protein) precludes or accommodates ERE-mediated transcription.

We are grateful to John A. Katzenellenbogen for providing the EDC compounds, to J. Larry Jameson for the ERα^NERKI/+ mice, to Andree Krust and Pierre Chambon for the ERα^+/− mice, and to Richard Lyttle and John A. Katzenellenbogen for critically reviewing the manuscript prior to its submission to the journal. We also thank Charles O'Brien and Robert S. Weinstein for helpful discussions, Verenda G. Lowe and Aaron D. Warren for technical assistance, and Robyn I. DeWall for assistance with the preparation of the manuscript.

This work was supported by the National Institutes of Health (P01 AG013918, R01 AR051187) and the Department of Veterans Affairs (Merit review and REAP).