MiR-1224-5p modulates osteogenesis by coordinating osteoblast/osteoclast differentiation via the Rap1 signaling target ADCY2

Abstract

MicroRNAs (miRNAs) broadly regulate normal biological functions of bone and the progression of fracture healing and osteoporosis. Recently, it has been reported that miR-1224-5p in fracture plasma is a potential therapy for osteogenesis. To investigate the roles of miR-1224-5p and the Rap1 signaling pathway in fracture healing and osteoporosis development and progression, we used BMMs, BMSCs, and skull osteoblast precursor cells for in vitro osteogenesis and osteoclastogenesis studies. Osteoblastogenesis and osteoclastogenesis were detected by ALP, ARS, and TRAP staining and bone slice resorption pit assays. The miR-1224-5p target gene was assessed by siRNA-mediated target gene knockdown and luciferase reporter assays. To explore the Rap1 pathway, we performed high-throughput sequencing, western blotting, RT-PCR, chromatin immunoprecipitation assays and immunohistochemical staining. In vivo, bone healing was judged by the cortical femoral defect, cranial bone defect and femoral fracture models. Progression of osteoporosis was evaluated by an ovariectomy model and an aged osteoporosis model. We discovered that the expression of miR-1224-5p was positively correlated with fracture healing progression. Moreover, in vitro, overexpression of miR-1224-5p slowed Rankl-induced osteoclast differentiation and promoted osteoblast differentiation via the Rap1-signaling pathway by targeting ADCY2. In addition, in vivo overexpression of miR-1224-5p significantly promoted fracture healing and ameliorated the progression of osteoporosis caused by estrogen deficiency or aging. Furthermore, knockdown of miRNA-1224-5p inhibited bone regeneration in mice and accelerated the progression of osteoporosis in elderly mice. Taken together, these results identify miR-1224-5p as a key bone osteogenic regulator, which may be a potential therapeutic target for osteoporosis and fracture nonunion.

circSLC25A13 acts as a ceRNA to regulate AML progression via miR‐616‐3p/ADCY2 axis

“…ADCY2 is a versatile regulator capable of playing diverse roles in disease. It has been reported that ADCY2 gene polymorphisms are involved in the genetic susceptibility to osteoporosis and fracture nonunion 21 . Estradiol promotes the expression of cAMP via miR‐148a/152‐ADCY2 to regulate the growth and function of mesenchymal stem cells 22 .…”

“…It has been reported that ADCY2 gene polymorphisms are involved in the genetic susceptibility to osteoporosis and fracture nonunion. 21 Estradiol promotes the expression of cAMP via miR-148a/152-ADCY2 to regulate the growth and function of mesenchymal stem cells. 22 Moreover, ADCY2 is associated with severe chronic obstructive pulmonary disease and lung function levels, 23 and ADCY2 interaction with air pollution is vital for asthma development.…”

Screening of Prognostic Molecular Markers and Establishment of Prognostic Model for G-protein Coupled Receptor-Related Genes in Epithelial Ovarian Serous Cancer Based on Machine Learning Method

“…The G protein-coupled receptor pathway is nowadays a hot spot in cancer immune research, and some members of GPCRs are actually demonstrated as having a role as prognostic factors in a variety of cancers [50][51][52].In KEGG enrichment analysis, the main enrichments were in the neuroligand-receptor interaction, phospholipase D signaling pathway, oxytocin signaling pathway, CGMP-PKG signaling pathway, and Rap1 signaling pathway. Among them, the RAP1 signaling pathway, which is regulated by other interacting genes in osteoporosis and colorectal cancer, mediates the value-added of tumor cells or osteoblasts [53,54].…”

Epigenetic regulation in metabolic diseases: mechanisms and advances in clinical study

“…254 miR-1224-5p is a vital bone osteogenic regulator and alleviates osteoporosis by targeting ADCY2 to promote osteoblast differentiation through the Rap1 signalling pathway and inhibiting RANKL-induced osteoclast differentiation. 255 miR-26a-5p is enriched in urine-derived stem cell extracellular vesicles (USCs-EVs). 256 By inhibiting HDAC4, miR-26a-5p derived from USCs-EVs can activate the HIF-1α and vascular endothelial growth factor A (VEGFA) pathway to facilitate the differentiation of osteogenic precursor cells, thus alleviating diabetic osteoporosis (DOP).…”