Bone resorption and bone formation appear to be tightly coupled. To explore osteoclast influences on osteoblast recruitment and differentiation, we examined osteoclast conditioned media (CM) for influences on osteoblasts. Osteoclast CM stimulated human mesenchymal stem (hMS) cell mineralized nodule formation. We identified candidate osteoclast-derived coupling factors using Affymetrix microarray. We observed induction of sphingosine kinase 1 (SPHK1), WntlOb, and BMP6 in mature multinucleated osteoclasts as compared to pre-osteoclasts. Stimulation of hMS cell nodule formation by the osteoclast CM was attenuated by the Wnt antagonist, Dkk1, a BMP-6 neutralizing antibody, and by a S1P antagonist. Sclerostin expression was elevated in osteoclast precursors and rapidly down-regulated during differentiation. CM from osteoclasts generated in vitro from 18-month old mice was unable to support hMS cell mineralization. TGF-beta treatment elevated SPHK1 and WntlOb expression. Our central hypothesis is that osteoclast production of SPHK1, WntlOb, and BMP6, combined with decreased Sclerostin expression, promotes osteoblast precursor recruitment, proliferation, differentiation, and survival and that each factor plays an integral role in maintaining coupling between osteoclast-mediated bone resorption and osteoblast-mediated bone formation.
In Aim 1, we will use functional cell assays and gene expression to ascertain the mechanisms by which osteoclast-derived coupling factors promote osteoblast gene expression, recruitment, proliferation, differentiation, and survival in vitro.
In Aim 2 we will use molecular approaches to determine the mechanisms by which Sclerostin, SPHK1, WntlOb, and BMP6 are modulated during osteoclast differentiation in vitro.
In Aim 3 we will use an in vivo model to examine the role of TGF-beta regulation of coupling factor production on osteoclast-mediated coupling of bone resorption to bone formation.
In Aim 4 we will examine gene expression and cellular assays to resolve the contribution of age in osteoclast support of osteoblast maturation and mineralization. Together, these studies will provide important novel information on how osteoclasts control osteoblast-mediated bone formation.

Public Health Relevance

A crucial question in bone biology is how osteoblasts are recruited to a resorption site and how the amount of bone laid down is controlled. Studies have implicated osteoclasts as important modulators of bone formation. Understanding the mechanisms by which osteoclasts influence osteoblast recruitment, differentiation, and survival will likely lead to more effective theapies to selectively promote bone formation.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Program Projects (P01)
Project #
5P01AG004875-30
Application #
8494474
Study Section
Special Emphasis Panel (ZAG1-ZIJ-8)
Project Start
Project End
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
30
Fiscal Year
2013
Total Cost
$266,554
Indirect Cost
$90,145
Name
Mayo Clinic, Rochester
Department
Type
DUNS #
006471700
City
Rochester
State
MN
Country
United States
Zip Code
55905
Drake, Matthew T; Khosla, Sundeep (2016) Hormonal and systemic regulation of sclerostin. Bone :
Rocca, Walter A; Gazzuola-Rocca, Liliana; Smith, Carin Y et al. (2016) Accelerated Accumulation of Multimorbidity After Bilateral Oophorectomy: A Population-Based Cohort Study. Mayo Clin Proc 91:1577-1589
McGee-Lawrence, Meghan E; Carpio, Lomeli R; Schulze, Ryan J et al. (2016) Hdac3 Deficiency Increases Marrow Adiposity and Induces Lipid Storage and Glucocorticoid Metabolism in Osteochondroprogenitor Cells. J Bone Miner Res 31:116-28
Farr, Joshua N; Khosla, Sundeep (2016) Determinants of bone strength and quality in diabetes mellitus in humans. Bone 82:28-34
Sellmeyer, Deborah E; Civitelli, Roberto; Hofbauer, Lorenz C et al. (2016) Skeletal Metabolism, Fracture Risk, and Fracture Outcomes in Type 1 and Type 2 Diabetes. Diabetes 65:1757-66
Nicks, Kristy M; Fujita, Koji; Fraser, Daniel et al. (2016) Deletion of Estrogen Receptor Beta in Osteoprogenitor Cells Increases Trabecular but Not Cortical Bone Mass in Female Mice. J Bone Miner Res 31:606-14
Schafer, Marissa J; Atkinson, Elizabeth J; Vanderboom, Patrick M et al. (2016) Quantification of GDF11 and Myostatin in Human Aging and Cardiovascular Disease. Cell Metab 23:1207-15
Weivoda, Megan M; Ruan, Ming; Hachfeld, Christine M et al. (2016) Wnt Signaling Inhibits Osteoclast Differentiation by Activating Canonical and Noncanonical cAMP/PKA Pathways. J Bone Miner Res 31:65-75
Weivoda, Megan M; Ruan, Ming; Pederson, Larry et al. (2016) Osteoclast TGF-β Receptor Signaling Induces Wnt1 Secretion and Couples Bone Resorption to Bone Formation. J Bone Miner Res 31:76-85
Roforth, Matthew M; Farr, Joshua N; Fujita, Koji et al. (2015) Global transcriptional profiling using RNA sequencing and DNA methylation patterns in highly enriched mesenchymal cells from young versus elderly women. Bone 76:49-57

Showing the most recent 10 out of 371 publications