Zoledronic acid promotes osteoclasts ferroptosis by inhibiting FBXO9-mediated p53 ubiquitination and degradation

Abstract

Bisphosphonates (BPs)-related osteonecrosis of jaw (BRONJ) is a severe complication of the long-term administration of BPs. The development of BRONJ is associated with the cell death of osteoclasts, but the underlying mechanism remains unclear. In the current study, the role of Zoledronic acid (ZA), a kind of bisphosphonates, in suppressing the growth of osteoclasts was investigated and its underlying mechanism was explored. The role of ZA in regulating osteoclasts function was evaluated in the RANKL-induced cell model. Cell viability was assessed by cell counting kit-8 (CCK-8) assay and fluorescein diacetate (FDA)-staining. We confirmed that ZA treatment suppressed cell viability of osteoclasts. Furthermore, ZA treatment led to osteoclasts death by facilitating osteoclasts ferroptosis, as evidenced by increased Fe2+, ROS, and malonyldialdehyde (MDA) level, and decreased glutathione peroxidase 4 (GPX4) and glutathione (GSH) level. Next, the gene expression profiles of alendronate- and risedronate-treated osteoclasts were obtained from Gene Expression Omnibus (GEO) dataset, and 18 differentially expressed genes were identified using venn diagram analysis. Among these 18 genes, the expression of F-box protein 9 (FBXO9) was inhibited by ZA treatment. Knockdown of FBXO9 resulted in osteoclasts ferroptosis. More important, FBXO9 overexpression repressed the effect of ZA on regulating osteoclasts ferroptosis. Mechanistically, FBXO9 interacted with p53 and decreased the protein stability of p53. Collectively, our study showed that ZA induced osteoclast cells ferroptosis by triggering FBXO9-mediated p53 ubiquitination and degradation.

FBXO9 Mediates the Cancer-Promoting Effects of ZNF143 by Degrading FBXW7 and Facilitates Drug Resistance in Hepatocellular Carcinoma

“…In recent years, F-box proteins have been identified as essential factors for malignancies via target degradation ( 12 , 28 ). FBXO9 is a mysterious member of the F-box protein family ( 16 ), and a few available studies have shown that FBXO9 is involved in the development of the nervous system or adipose tissue ( 17 , 18 ), in pathological changes in the muscle in diabetes ( 17 ) and epithelial damage in respiratory distress syndrome ( 18 ), and in the adverse effects of zoledronic acid ( 19 ) or as a target of sepsis ( 18 ), and participates in disease progression in hematological tumors by regulating the activity of the mTOR pathway ( 20 ) and the proteasome ( 8 ). However, the biological functions and mechanisms underlying the role of FBXO9 in HCC have not been reported.…”

“…However, less attention has been paid to FBXO9 ( 16 ). To date, several substrates of FBXO9 have been identified, including Neurog2 ( 17 ), PRMT4 ( 18 ), p53 ( 19 ), PPARγ, BK channel β1 subunit, TEL2, and TTI1 ( 17 , 20 ). FBXO9 participates in several cellular processes by degrading these substrates.…”

The Regulatory Role of Ferroptosis in Bone Homeostasis

“…Iron ions is capable to induce OC differentiation and bone resorption through the production of ROS [ 48 , 49 ]. Zoledronic acid (ZA), a bisphosphonate, has been reported to inhibit OC growth via induction of ferroptosis of the OC [ 49 ].…”

“…Iron ions is capable to induce OC differentiation and bone resorption through the production of ROS [ 48 , 49 ]. Zoledronic acid (ZA), a bisphosphonate, has been reported to inhibit OC growth via induction of ferroptosis of the OC [ 49 ]. The role of ZA in regulating osteoclast function was evaluated using a RANKL-induced cell model, which indicated that ZA treatment suppressed the cell viability of osteoclasts and facilitated osteoclast ferroptosis with an increase in iron ions and ROS and decrease in the GPX4 and GSH level [ 49 ].…”

“…Zoledronic acid (ZA), a bisphosphonate, has been reported to inhibit OC growth via induction of ferroptosis of the OC [ 49 ]. The role of ZA in regulating osteoclast function was evaluated using a RANKL-induced cell model, which indicated that ZA treatment suppressed the cell viability of osteoclasts and facilitated osteoclast ferroptosis with an increase in iron ions and ROS and decrease in the GPX4 and GSH level [ 49 ]. Similarly, ferroptosis was reported involved in OC function during RANKL-induced differentiation and is induced by iron starvation response and ferritin phagocytosis [ 50 ].…”