Myeloid lineage ablation of phlpp1 regulates m-csf signaling and tempers bone resorption in female mice

Ismael Y. Karkache, Jeyaram R. Damodaran, David H.H. Molstad, Kim C. Mansky, Elizabeth W. Bradley

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Prior work demonstrated that Phlpp1 deficiency alters trabecular bone mass and enhances M-CSF responsiveness, but the cell types and requirement of Phlpp1 for this effect were unclear. To understand the function of Phlpp1 within myeloid lineage cells, we crossed Phlpp1 floxed mice with mice harboring LysM-Cre. Micro-computed tomography of the distal femur of 12-week-old mice revealed a 30% increase in bone volume per total volume of Phlpp1 female conditional knockouts, but we did not observe significant changes within male Phlpp1 cKOLysM mice. Bone histomorphmetry of the proximal tibia further revealed that Phlpp1 cKOLysM females exhibited elevated osteoclast numbers, but conversely had reduced levels of serum markers of bone resorption as compared to littermate controls. Osteoblast number and serum markers of bone formation were unchanged. In vitro assays confirmed that Phlpp1 ablation enhanced osteoclast number and area, but limited bone resorption. Additionally, reconstitution with exogenous Phlpp1 suppressed osteoclast numbers. Dose response assays demonstrated that Phlpp1−/− cells are more responsive to M-CSF, but reconstitution with Phlpp1 abrogated this effect. Furthermore, small molecule-mediated Phlpp inhibition enhanced osteoclast numbers and size. Enhanced phosphorylation of Phlpp substrates—including Akt, ERK1/2, and PKCζ—accompanied these observations. In contrast, actin cytoskeleton disruption occurred within Phlpp inhibitor treated osteoclasts. Moreover, Phlpp inhibition reduced resorption of cells cultured on bovine bone slices in vitro. Our results demonstrate that Phlpp1 deficiency within myeloid lineage cells enhances bone mass by limiting bone resorption while leaving osteoclast numbers intact; moreover, we show that Phlpp1 represses osteoclastogenesis and controls responses to M-CSF.

Original languageEnglish (US)
Article number9702
JournalInternational journal of molecular sciences
Volume22
Issue number18
DOIs
StatePublished - Sep 2021

Bibliographical note

Funding Information:
Acknowledgments: This work was made possible by research grants from the National Institutes of Health (AR072634), and the University of Minnesota Board of Regents. These contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH.

Funding Information:
Funding: This research was funded by the National Institute of Health, National Institute for Arthritis, Musculoskeletal and Skin Diseases, grant number AR072634 and the University of Minnesota.

Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Keywords

  • Akt
  • Bone mass
  • ERK
  • MEK
  • Osteoclast
  • Osteoporosis
  • PDZ domain
  • PH domain
  • Protein Kinase C (PKC)
  • Ras-association domain
  • Sexual dimorphism

PubMed: MeSH publication types

  • Journal Article

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