microRNA-324 mediates bone homeostasis and the regulation of osteoblast and osteoclast differentiation and activity

Bone. 2025 Jan:190:117273. doi: 10.1016/j.bone.2024.117273. Epub 2024 Oct 9.

Abstract

MicroRNAs (miRNAs) modulate the expression of other RNA molecules. One miRNA can target many transcripts, allowing each miRNA to play key roles in many biological pathways. Defects in bone homeostasis result in common age-related diseases including osteoporosis. Serum levels of miR-324-3p positively correlate with several features of bone maintenance. In contrast here, using in vivo micro-computed tomography and histology, global miR-324-null mice demonstrated increased bone mineral density and both trabecular and cortical thickness, with effect magnitudes increasing with age. The bone marrow of miR-324-null mice had reduced lipid content while TRAP staining revealed a decrease in osteoclasts, with histomorphometry demonstrating an increased rate of bone formation. Ex vivo assays showed that the high bone mass phenotype of miR-324-null mice resulted from both increased osteoblast activity and decreased osteoclastogenesis. RNA-seq analysis of osteoblasts, osteoclasts and bone marrow macrophages and target validation assays identified that the osteoclast fusion regulator Pin1 and the master osteogenic regulator Runx2 were targets of miR-324-5p in osteoclast lineage cells and osteoblasts, respectively. Indeed, in vitro Runx2 overexpression recapitulated the increased osteogenesis and decreased adipogenesis phenotype observed in vivo by the loss of miR-324. Overall, these data demonstrate the importance of miR-324 in bone homeostasis by regulating aspects of both bone formation and remodelling. Elucidation of pathways regulated by miR-324 offer promise for the treatment of bone diseases such as osteoporosis.

Keywords: Bone; Osteoblast; Osteoclast; microRNA.

MeSH terms

  • Animals
  • Bone Density
  • Bone and Bones* / metabolism
  • Cell Differentiation*
  • Core Binding Factor Alpha 1 Subunit / metabolism
  • Homeostasis*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Osteoblasts* / metabolism
  • Osteoclasts* / metabolism
  • Osteogenesis* / genetics
  • Osteogenesis* / physiology
  • X-Ray Microtomography

Substances

  • MicroRNAs
  • Core Binding Factor Alpha 1 Subunit