Estrogens antagonize RUNX2-mediated osteoblast-driven osteoclastogenesis through regulating RANKL membrane association

Anthony Martin, Jian Xiong, Theodora Koromila, Jie S. Ji, Stephanie Chang, Yae S. Song, Jonathan L. Miller, Chun Ya Han, Paul Kostenuik, Susan Miranda, Nyam Osor Chimge, Yankel Gabet, Baruch Frenkel

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Abstract

In addition to its thoroughly investigated role in bone formation, the osteoblast master transcription factor RUNX2 also promotes osteoclastogenesis and bone resorption. Here we demonstrate that 17β-estradiol (E2), strongly inhibits RUNX2-mediated osteoblast-driven osteoclastogenesis in co-cultures. Towards deciphering the underlying mechanism, we induced premature expression of RUNX2 in primary murine pre-osteoblasts, which resulted in robust differentiation of co-cultured splenocytes into mature osteoclasts. This was attributable to RUNX2-mediated increase in RANKL secretion, determined by ELISA, as well as to RUNX2-mediated increase in RANKL association with the osteoblast membrane, demonstrated using confocal fluorescence microscopy. The increased association with the osteoblast membrane was recapitulated by transiently expressed GFP-RANKL. E2 abolished the RUNX2-mediated increase in membrane-associated RANKL and GFP-RANKL, as well as the concomitant osteoclastogenesis. RUNX2-mediated RANKL cellular redistribution was attributable in part to a decrease in Opg expression, but E2 did not influence Opg expression either in the presence or absence of RUNX2. Diminution of RUNX2-mediated osteoclastogenesis by E2 occurred regardless of whether the pre-osteoclasts were derived from wild type or estrogen receptor alpha (ERα)-knockout mice, suggesting that activated ERα inhibited osteoblast-driven osteoclastogenesis by acting in osteoblasts, possibly targeting RUNX2. Indeed, microarray analysis demonstrated global attenuation of the RUNX2 response by E2, including abrogation of Pstpip2 expression, which likely plays a critical role in membrane trafficking. Finally, the selective ER modulators (SERMs) tamoxifen and raloxifene mimicked E2 in abrogating the stimulatory effect of osteoblastic RUNX2 on osteoclast differentiation in the co-culture assay. Thus, E2 antagonizes RUNX2-mediated RANKL trafficking and subsequent osteoclastogenesis. Targeting RUNX2 and/or downstream mechanisms that regulate RANKL trafficking may lead to the development of improved SERMs and possibly non-hormonal therapeutic approaches to high turnover bone disease.

Original languageEnglish (US)
Pages (from-to)96-104
Number of pages9
JournalBone
Volume75
DOIs
StatePublished - Jun 1 2015

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Osteoblasts
Osteogenesis
Estrogens
Membranes
Estrogen Receptor alpha
Osteoclasts
Coculture Techniques
Selective Estrogen Receptor Modulators
Bone Diseases
Tamoxifen
Microarray Analysis
Bone Resorption
Fluorescence Microscopy
Knockout Mice
Confocal Microscopy
Estradiol
Transcription Factors
Enzyme-Linked Immunosorbent Assay

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Physiology
  • Histology

Cite this

Martin, A., Xiong, J., Koromila, T., Ji, J. S., Chang, S., Song, Y. S., ... Frenkel, B. (2015). Estrogens antagonize RUNX2-mediated osteoblast-driven osteoclastogenesis through regulating RANKL membrane association. Bone, 75, 96-104. https://doi.org/10.1016/j.bone.2015.02.007

Estrogens antagonize RUNX2-mediated osteoblast-driven osteoclastogenesis through regulating RANKL membrane association. / Martin, Anthony; Xiong, Jian; Koromila, Theodora; Ji, Jie S.; Chang, Stephanie; Song, Yae S.; Miller, Jonathan L.; Han, Chun Ya; Kostenuik, Paul; Miranda, Susan; Chimge, Nyam Osor; Gabet, Yankel; Frenkel, Baruch.

In: Bone, Vol. 75, 01.06.2015, p. 96-104.

Research output: Contribution to journalArticle

Martin, A, Xiong, J, Koromila, T, Ji, JS, Chang, S, Song, YS, Miller, JL, Han, CY, Kostenuik, P, Miranda, S, Chimge, NO, Gabet, Y & Frenkel, B 2015, 'Estrogens antagonize RUNX2-mediated osteoblast-driven osteoclastogenesis through regulating RANKL membrane association', Bone, vol. 75, pp. 96-104. https://doi.org/10.1016/j.bone.2015.02.007
Martin, Anthony ; Xiong, Jian ; Koromila, Theodora ; Ji, Jie S. ; Chang, Stephanie ; Song, Yae S. ; Miller, Jonathan L. ; Han, Chun Ya ; Kostenuik, Paul ; Miranda, Susan ; Chimge, Nyam Osor ; Gabet, Yankel ; Frenkel, Baruch. / Estrogens antagonize RUNX2-mediated osteoblast-driven osteoclastogenesis through regulating RANKL membrane association. In: Bone. 2015 ; Vol. 75. pp. 96-104.
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