Estrogens and androgens inhibit association of RANKL with the pre-osteoblast membrane through post-translational mechanisms

Anthony Martin, Jiali Yu, Jian Xiong, Aysha B. Khalid, Benita Katzenellenbogen, Sung Hoon Kim, John A. Katzenellenbogen, Suchinda Malaivijitnond, Yankel Gabet, Susan A. Krum, Baruch Frenkel

Research output: Contribution to journalArticle

Abstract

We have recently demonstrated that RUNX2 promoted, and 17β-Estradiol (E2) diminished, association of RANKL with the cell membrane in pre-osteoblast cultures. Here we show that, similar to E2, dihydrotestosterone (DHT) diminishes association of RANKL, and transiently transfected GFP-RANKL with the pre-osteoblast membrane without decreasing total RANKL mRNA or protein levels. Diminution of membrane-associated RANKL was accompanied with marked suppression of osteoclast differentiation from co-cultured pre-osteoclasts, even though DHT increased, not decreased, RANKL concentrations in pre-osteoblast conditioned media. A marked decrease in membrane-associated RANKL was observed after 30 min of either E2 or DHT treatment, and near-complete inhibition was observed by 1 hr, suggesting that the diminution of RANKL membrane association was mediated through non-genomic mechanisms. Further indicating dispensability of nuclear action of estrogen receptor, E2-mediated inhibition of RANKL membrane association was mimicked by an estrogen dendrimer conjugate (EDC) that cannot enter the cell nucleus. Finally, the inhibitory effect of E2 and DHT on RANKL membrane association was counteracted by the MMP inhibitor NNGH, and the effect of E2 (and not DHT) was antagonized by the Src inhibitor SU6656. Taken together, these results suggest that estrogens and androgens inhibit osteoblast-driven osteoclastogenesis through non-genomic mechanism(s) that entail, MMP-mediated RANKL dissociation from the cell membrane.

Original languageEnglish (US)
Pages (from-to)3798-3807
Number of pages10
JournalJournal of Cellular Physiology
Volume232
Issue number12
DOIs
StatePublished - Dec 2017

Fingerprint

Osteoblasts
Dihydrotestosterone
Androgens
Estrogens
Association reactions
Membranes
Osteoclasts
Cell membranes
Cell Membrane
Dendrimers
Matrix Metalloproteinase Inhibitors
Conditioned Culture Medium
Cell Nucleus
Matrix Metalloproteinases
Cell culture
Osteogenesis
Estrogen Receptors
Estradiol
Cells
Messenger RNA

Keywords

  • MMP
  • RANKL presentation
  • Src
  • membrane-initiated estrogen signaling
  • sex steroids

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Estrogens and androgens inhibit association of RANKL with the pre-osteoblast membrane through post-translational mechanisms. / Martin, Anthony; Yu, Jiali; Xiong, Jian; Khalid, Aysha B.; Katzenellenbogen, Benita; Kim, Sung Hoon; Katzenellenbogen, John A.; Malaivijitnond, Suchinda; Gabet, Yankel; Krum, Susan A.; Frenkel, Baruch.

In: Journal of Cellular Physiology, Vol. 232, No. 12, 12.2017, p. 3798-3807.

Research output: Contribution to journalArticle

Martin, A, Yu, J, Xiong, J, Khalid, AB, Katzenellenbogen, B, Kim, SH, Katzenellenbogen, JA, Malaivijitnond, S, Gabet, Y, Krum, SA & Frenkel, B 2017, 'Estrogens and androgens inhibit association of RANKL with the pre-osteoblast membrane through post-translational mechanisms', Journal of Cellular Physiology, vol. 232, no. 12, pp. 3798-3807. https://doi.org/10.1002/jcp.25862
Martin, Anthony ; Yu, Jiali ; Xiong, Jian ; Khalid, Aysha B. ; Katzenellenbogen, Benita ; Kim, Sung Hoon ; Katzenellenbogen, John A. ; Malaivijitnond, Suchinda ; Gabet, Yankel ; Krum, Susan A. ; Frenkel, Baruch. / Estrogens and androgens inhibit association of RANKL with the pre-osteoblast membrane through post-translational mechanisms. In: Journal of Cellular Physiology. 2017 ; Vol. 232, No. 12. pp. 3798-3807.
@article{776d45bfd13f4ed3ae2de596b91b1638,
title = "Estrogens and androgens inhibit association of RANKL with the pre-osteoblast membrane through post-translational mechanisms",
abstract = "We have recently demonstrated that RUNX2 promoted, and 17β-Estradiol (E2) diminished, association of RANKL with the cell membrane in pre-osteoblast cultures. Here we show that, similar to E2, dihydrotestosterone (DHT) diminishes association of RANKL, and transiently transfected GFP-RANKL with the pre-osteoblast membrane without decreasing total RANKL mRNA or protein levels. Diminution of membrane-associated RANKL was accompanied with marked suppression of osteoclast differentiation from co-cultured pre-osteoclasts, even though DHT increased, not decreased, RANKL concentrations in pre-osteoblast conditioned media. A marked decrease in membrane-associated RANKL was observed after 30 min of either E2 or DHT treatment, and near-complete inhibition was observed by 1 hr, suggesting that the diminution of RANKL membrane association was mediated through non-genomic mechanisms. Further indicating dispensability of nuclear action of estrogen receptor, E2-mediated inhibition of RANKL membrane association was mimicked by an estrogen dendrimer conjugate (EDC) that cannot enter the cell nucleus. Finally, the inhibitory effect of E2 and DHT on RANKL membrane association was counteracted by the MMP inhibitor NNGH, and the effect of E2 (and not DHT) was antagonized by the Src inhibitor SU6656. Taken together, these results suggest that estrogens and androgens inhibit osteoblast-driven osteoclastogenesis through non-genomic mechanism(s) that entail, MMP-mediated RANKL dissociation from the cell membrane.",
keywords = "MMP, RANKL presentation, Src, membrane-initiated estrogen signaling, sex steroids",
author = "Anthony Martin and Jiali Yu and Jian Xiong and Khalid, {Aysha B.} and Benita Katzenellenbogen and Kim, {Sung Hoon} and Katzenellenbogen, {John A.} and Suchinda Malaivijitnond and Yankel Gabet and Krum, {Susan A.} and Baruch Frenkel",
year = "2017",
month = "12",
doi = "10.1002/jcp.25862",
language = "English (US)",
volume = "232",
pages = "3798--3807",
journal = "Journal of Cellular Physiology",
issn = "0021-9541",
publisher = "Wiley-Liss Inc.",
number = "12",

}

TY - JOUR

T1 - Estrogens and androgens inhibit association of RANKL with the pre-osteoblast membrane through post-translational mechanisms

AU - Martin, Anthony

AU - Yu, Jiali

AU - Xiong, Jian

AU - Khalid, Aysha B.

AU - Katzenellenbogen, Benita

AU - Kim, Sung Hoon

AU - Katzenellenbogen, John A.

AU - Malaivijitnond, Suchinda

AU - Gabet, Yankel

AU - Krum, Susan A.

AU - Frenkel, Baruch

PY - 2017/12

Y1 - 2017/12

N2 - We have recently demonstrated that RUNX2 promoted, and 17β-Estradiol (E2) diminished, association of RANKL with the cell membrane in pre-osteoblast cultures. Here we show that, similar to E2, dihydrotestosterone (DHT) diminishes association of RANKL, and transiently transfected GFP-RANKL with the pre-osteoblast membrane without decreasing total RANKL mRNA or protein levels. Diminution of membrane-associated RANKL was accompanied with marked suppression of osteoclast differentiation from co-cultured pre-osteoclasts, even though DHT increased, not decreased, RANKL concentrations in pre-osteoblast conditioned media. A marked decrease in membrane-associated RANKL was observed after 30 min of either E2 or DHT treatment, and near-complete inhibition was observed by 1 hr, suggesting that the diminution of RANKL membrane association was mediated through non-genomic mechanisms. Further indicating dispensability of nuclear action of estrogen receptor, E2-mediated inhibition of RANKL membrane association was mimicked by an estrogen dendrimer conjugate (EDC) that cannot enter the cell nucleus. Finally, the inhibitory effect of E2 and DHT on RANKL membrane association was counteracted by the MMP inhibitor NNGH, and the effect of E2 (and not DHT) was antagonized by the Src inhibitor SU6656. Taken together, these results suggest that estrogens and androgens inhibit osteoblast-driven osteoclastogenesis through non-genomic mechanism(s) that entail, MMP-mediated RANKL dissociation from the cell membrane.

AB - We have recently demonstrated that RUNX2 promoted, and 17β-Estradiol (E2) diminished, association of RANKL with the cell membrane in pre-osteoblast cultures. Here we show that, similar to E2, dihydrotestosterone (DHT) diminishes association of RANKL, and transiently transfected GFP-RANKL with the pre-osteoblast membrane without decreasing total RANKL mRNA or protein levels. Diminution of membrane-associated RANKL was accompanied with marked suppression of osteoclast differentiation from co-cultured pre-osteoclasts, even though DHT increased, not decreased, RANKL concentrations in pre-osteoblast conditioned media. A marked decrease in membrane-associated RANKL was observed after 30 min of either E2 or DHT treatment, and near-complete inhibition was observed by 1 hr, suggesting that the diminution of RANKL membrane association was mediated through non-genomic mechanisms. Further indicating dispensability of nuclear action of estrogen receptor, E2-mediated inhibition of RANKL membrane association was mimicked by an estrogen dendrimer conjugate (EDC) that cannot enter the cell nucleus. Finally, the inhibitory effect of E2 and DHT on RANKL membrane association was counteracted by the MMP inhibitor NNGH, and the effect of E2 (and not DHT) was antagonized by the Src inhibitor SU6656. Taken together, these results suggest that estrogens and androgens inhibit osteoblast-driven osteoclastogenesis through non-genomic mechanism(s) that entail, MMP-mediated RANKL dissociation from the cell membrane.

KW - MMP

KW - RANKL presentation

KW - Src

KW - membrane-initiated estrogen signaling

KW - sex steroids

UR - http://www.scopus.com/inward/record.url?scp=85019125488&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85019125488&partnerID=8YFLogxK

U2 - 10.1002/jcp.25862

DO - 10.1002/jcp.25862

M3 - Article

C2 - 28213978

AN - SCOPUS:85019125488

VL - 232

SP - 3798

EP - 3807

JO - Journal of Cellular Physiology

JF - Journal of Cellular Physiology

SN - 0021-9541

IS - 12

ER -