Identifying therapeutic biomarkers of zoledronic acid by metabolomics

Xiang Li; Zi-Yuan Wang; Na Ren; Zhan-Ying Wei; Wei-Wei Hu; Jie-Mei Gu; Zhen-Lin Zhang; Xiang-Tian Yu; Chun Wang

doi:10.3389/fphar.2023.1084453

Identifying therapeutic biomarkers of zoledronic acid by metabolomics

Front Pharmacol. 2023 Apr 25:14:1084453. doi: 10.3389/fphar.2023.1084453. eCollection 2023.

Authors

Xiang Li¹, Zi-Yuan Wang¹, Na Ren¹, Zhan-Ying Wei¹, Wei-Wei Hu¹, Jie-Mei Gu¹, Zhen-Lin Zhang¹, Xiang-Tian Yu², Chun Wang¹

Affiliations

¹ Department of Osteoporosis and Bone Diseases, Shanghai Clinical Research Center of Bone Disease, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.
² Clinical Research Center, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.

Abstract

Zoledronic acid (ZOL) is a potent antiresorptive agent that increases bone mineral density (BMD) and reduces fracture risk in postmenopausal osteoporosis (PMOP). The anti-osteoporotic effect of ZOL is determined by annual BMD measurement. In most cases, bone turnover markers function as early indicators of therapeutic response, but they fail to reflect long-term effects. We used untargeted metabolomics to characterize time-dependent metabolic shifts in response to ZOL and to screen potential therapeutic markers. In addition, bone marrow RNA-seq was performed to support plasma metabolic profiling. Sixty rats were assigned to sham-operated group (SHAM, n = 21) and ovariectomy group (OVX, n = 39) and received sham operation or bilateral ovariectomy, respectively. After modeling and verification, rats in the OVX group were further divided into normal saline group (NS, n = 15) and ZOL group (ZA, n = 18). Three doses of 100 μg/kg ZOL were administrated to the ZA group every 2 weeks to simulate 3-year ZOL therapy in PMOP. An equal volume of saline was administered to the SHAM and NS groups. Plasma samples were collected at five time points for metabolic profiling. At the end of the study, selected rats were euthanatized for bone marrow RNA-seq. A total number of 163 compound were identified as differential metabolites between the ZA and NS groups, including mevalonate, a critical molecule in target pathway of ZOL. In addition, prolyl hydroxyproline (PHP), leucyl hydroxyproline (LHP), 4-vinylphenol sulfate (4-VPS) were identified as differential metabolites throughout the study. Moreover, 4-VPS negatively correlated with increased vertebral BMD after ZOL administration as time-series analysis revealed. Bone marrow RNA-seq showed that the PI3K-AKT signaling pathway was significantly associated with ZOL-mediated changes in expression (adjusted-p = 0.018). In conclusion, mevalonate, PHP, LHP, and 4-VPS are candidate therapeutic markers of ZOL. The pharmacological effect of ZOL likely occurs through inhibition of the PI3K-AKT signaling pathway.

Keywords: RNA-Seq; biomarkers; metabolomics; osteoporosis; ovariectomy; rats; zoledronic acid.

Grants and funding

This work was supported by the National Key Research and Development Program of China (2021YFC2501705) and the National Natural Science Foundation of China (82070903, 82170895, and 82270933).