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miR-433-3p suppresses bone formation and mRNAs critical for osteoblast function in mice

Garcia, John, Smith, Spenser S., Karki, Sangita, Drissi, Hicham, Hrdlicka, Henry H., Youngstrom, Daniel W., Delany, Anne M.·2020·Journal of Bone and Mineral Research
DOI 10.1002/jbmr.4339
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Abstract

MicroRNAs (miRNAs) are key posttranscriptional regulators of osteoblastic commitment and differentiation. miR-433-3p was previously shown to target Runt-related transcription factor 2 (Runx2) and to be repressed by bone morphogenetic protein (BMP) signaling. Here, we show that miR-433-3p is progressively decreased during osteoblastic differentiation of primary mouse bone marrow stromal cells in vitro, and we confirm its negative regulation of this process. Although repressors of osteoblastic differentiation often promote adipogenesis, inhibition of miR-433-3p did not affect adipocyte differentiation in vitro. Multiple pathways regulate osteogenesis. Using luciferase-3′ untranslated region (UTR) reporter assays, five novel miR-433-3p targets involved in parathyroid hormone (PTH), mitogen-activated protein kinase (MAPK), Wnt, and glucocorticoid signaling pathways were validated. We show that Creb1 is a miR-433-3p target, and this transcription factor mediates key signaling downstream of PTH receptor activation. We also show that miR-433-3p targets hydroxysteroid 11-β dehydrogenase 1 (Hsd11b1), the enzyme that locally converts inactive glucocorticoids to their active form. miR-433-3p dampens glucocorticoid signaling, and targeting of Hsd11b1 could contribute to this phenomenon. Moreover, miR-433-3p targets R-spondin 3 (Rspo3), a leucine-rich repeat-containing G-protein coupled receptor (LGR) ligand that enhances Wnt signaling. Notably, Wnt canonical signaling is also blunted by miR-433-3p activity. In vivo, expression of a miR-433-3p inhibitor or tough decoy in the osteoblastic lineage increased trabecular bone volume. Mice expressing the miR-433-3p tough decoy displayed increased bone formation without alterations in osteoblast or osteoclast numbers or surface, indicating that miR-433-3p decreases osteoblast activity. Overall, we showed that miR-433-3p is a negative regulator of bone formation in vivo, targeting key bone-anabolic pathways including those involved in PTH signaling, Wnt, and endogenous glucocorticoids. Local delivery of miR-433-3p inhibitor could present a strategy for the management of bone loss disorders and bone defect repair. © 2021 American Society for Bone and Mineral Research (ASBMR).

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Metadata

Authors
Garcia, J., Smith, S. S., Karki, S., Drissi, H., Hrdlicka, H. H., Youngstrom, D. W., & Delany, A. M.
Affiliations
  • Center for Molecular Oncology UConn Health Farmington Connecticut USA
  • Department of Orthopedic Surgery University of California San Francisco San Francisco California USA
  • Department of Orthopaedics Emory University and Atlanta VA Medical Center Decatur Georgia USA
  • Gaylord Specialty Healthcare Wallingford Connecticut USA
  • Department of Orthopedic Surgery UConn Health Farmington Connecticut USA
ORCID
Venue
Journal of Bone and Mineral Research
Publisher
Oxford University Press (OUP)
Published
2020-12-01
Volume
36
Issue
9
Pages
1808-1822
ISSN
  • 0884-0431
  • 1523-4681
DOI
10.1002/jbmr.4339
Cited by
16
References
5
Language
en
Licence
Funders
  • American Society for Bone and Mineral Research Grant-in-Aid Program
  • Center for Molecular Oncology at UConn Health
  • Center for Scientific Review (AR044877, AR077962)
  • Connecticut Convergence Institute for Translation in Regenerative Engineering
  • UConn Health Center Research Advisory Council

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