Longitudinal changes of circulating miRNAs during bisphosphonate and teriparatide treatment in an animal model of postmenopausal osteoporosis

Weigl, Moritz<sup>1,2</sup>; Kocijan, Roland<sup>3,4,5</sup>; Ferguson, James<sup>2,4</sup>; Leinfellner, Gabriele<sup>2,4</sup>; Heimel, Patrick<sup>4,6</sup>; Feichtinger, Xaver<sup>4</sup>; Pietschmann, Peter<sup>7</sup>; Grillari, Johannes<sup>2,4,8</sup>; Zwerina, Jochen<sup>3</sup>; Redl, Heinz<sup>2,4</sup>; Hackl, Matthias<sup>1,2</sup>

Affiliations

¹TAmiRNA GmbH, Vienna, Austria

²Austrian Cluster for Tissue Regeneration, Vienna, Austria

³Ludwig Boltzmann Institute of Osteology at Hanusch Hospital of OEGK and AUVA Trauma Centre Meidling, Vienna, Austria

⁴Ludwig Boltzmann Institute for Experimental and Clinical Traumatology in AUVA Research Center, Vienna, Austria

⁵Medical Faculty of Bone Diseases, Sigmund Freud University, Vienna, Austria

⁶Karl Donath Laboratory for Hard Tissue and Biomaterial Research, University Clinic of Dentistry, Medical University of Vienna, Vienna, Austria

⁷Institute of Pathophysiology and Allergy Research, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria

⁸Institute of Molecular Biotechnology, Department of Biotechnology, BOKU - University of Natural Resources and Life Sciences Vienna, Vienna, Austria

Abstract and keywords

MicroRNAs regulate bone homeostasis, and circulating microRNAs have been proposed as novel bone biomarkers. The effect of anti-osteoporotic treatment on circulating microRNAs has not been described in detail. Therefore, we performed a comprehensive analysis of microRNA serum levels in ovariectomized (OVX) and sham-operated (SHAM) rats over 12 weeks of antiresorptive or osteoanabolic treatment. Forty-two Sprague Dawley rats underwent SHAM surgery (n = 10) or ovariectomy (n = 32). After 8 weeks, OVX rats were randomized to antiresorptive treatment with zoledronate (n = 11), osteoanabolic treatment with teriparatide (n = 11), or vehicle treatment (n = 10). Serum samples were collected at weeks 8, 12, 16, and 20 after surgery. A total of 91 microRNAs were analyzed by RT-qPCR in serum samples collected at week 20. Based on the results, 29 microRNAs were selected for longitudinal analysis at all four study time points. Changes in bone mineral density and microstructure were followed up by in vivo micro-CT and ex vivo nano-CT. Ovariectomy resulted in the loss of trabecular bone, which was reversed by osteoanabolic and antiresorptive treatment. Differential expression analysis identified 11 circulating miRNAs that were significantly regulated after treatment. For example, miR-107 and miR-31-5p increased in vehicle-treated OVX animals, whereas they decreased during teriparatide treatment. Additional miRNAs were identified that showed significant correlations to bone microstructure or bone miRNA expression, including miR-203a-3p, which exhibited a significant negative correlation to vertebral and tibial trabecular bone volume fraction (%). Longitudinal analysis confirmed eight microRNAs with significant changes in serum over time that were prevented by teriparatide and zoledronate treatment (miR-34a-5p, miR-31-5p, miR-30d-3p, miR-378a-5p) or teriparatide treatment only (miR-375-3p, miR-183-5p, miR-203a-3p, miR-203b-3p). Gene target network analysis identified WNT and Notch signaling as the main signaling pathways controlled by these miRNAs. Thus, ovariectomy results in time-dependent deregulation of circulating miRNAs compared with SHAM animals. Anti-osteoporotic treatments can rescue this effect, showing that bone-related miRNAs might act as novel biomarkers for treatment monitoring.

Keywords: BONE MICROSTRUCTURE; miRNAs; OSTEOPOROSIS; POSTMENOPAUSAL OSTEOPOROSIS; TERIPARATIDE; ZOLEDRONIC ACID

Links

DOI: 10.1002/jbmr.4276

PubMed: 33598975

WoS: 000627040500001

History

Received: 2020-12-15

Revised: 2020-02-05

Accepted: 2021-02-14

Online: 2021-02-17

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Cited By (1)

Year	Entry
2022	Heilmeier U, Hackl M, Schroeder F, Torabi S, Kapoor P, Vierlinger K, Eiriksdottir G, Gudmundsson EF, Harris TB, Gudnason V, Link TM, Grillari J, Schwartz AV. Circulating serum microRNAs including senescent miR-31-5p are associated with incident fragility fractures in older postmenopausal women with type 2 diabetes mellitus. Bone. May 2022;158:116308.