MicroRNA levels in bone and blood change during bisphosphonate and teriparatide therapy in an animal model of postmenopausal osteoporosis

Kocijan, Roland; Weigl, Moritz; Skalicky, Susanna; Geiger, Elisabeth; Ferguson, James; Leinfellner, Gabriele; Heimel, Patrick; Pietschmann, Peter; Grillari, Johannes; Redl, Heinz; Hackl, Matthias

Affiliations

¹Ludwig Boltzmann Institute for Experimental and Clinical Traumatology in AUVA Research Center, Donaueschingenstraße 13, 1200 Vienna, Austria

²Hanusch Hospital, 1st Medical Department, Heinrich Collin-Str. 30, 1140 Vienna, Austria

³TAmiRNA GmbH, Leberstrasse 20, 1110 Vienna, Austria

⁴Austrian Cluster for Tissue Regeneration, Austria

⁵Karl Donath Laboratory for Hard Tissue and Biomaterial Research, Department of Oral Surgery, University Clinic of Dentistry, Austria

⁶Department of Pathophysiology and Allergy Research, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria

⁷Christian Doppler Laboratory on Biotechnology of Skin Aging, Department of Biotechnology, BOKU - University of Natural Resources and Life Sciences Vienna, Vienna, Austria

Abstract and keywords

MicroRNAs control the activity of a variety of genes that are pivotal to bone metabolism. Therefore, the clinical utility of miRNAs as biomarkers and drug targets for bone diseases certainly merits further investigation. This study describes the use of an animal model of postmenopausal osteoporosis to generate a comprehensive dataset on miRNA regulation in bone tissue and peripheral blood during bone loss and specifically anti-resorptive and osteo-anabolic treatment.

Forty-two Sprague-Dawley rats were randomized to SHAM surgery (n = 10) or ovariectomy (OVX, n = 32). Eight weeks after surgery, OVX animals were further randomized to anti-resorptive treatment with zoledronate (n = 11), osteo-anabolic treatment with teriparatide (n = 11), or vehicle treatment (n = 10). After 12 weeks of treatment, bone and serum samples were used for microRNA analysis using next-generation sequencing (NGS), mRNA levels using RT-qPCR, and bone microarchitecture analysis using nanoCT.

Ovariectomy resulted in loss of trabecular bone, which was fully rescued using osteo-anabolic treatment, and partially rescued using anti-resorptive treatment. NGS revealed that both, anti-resorptive and anabolic treatment had a significant impact on miRNA levels in bone tissue and serum: out of 426 detected miRNAs, 46 miRNAs were regulated by teriparatide treatment an d 10 by zoledronate treatment (p-adj. < 0.1). Interestingly, teriparatide and zoledronate treatment were able to revert miRNA changes in tissue and serum of untreated OVX animals, such as the up-regulation of miR-203a-3p, a known osteo-inhibitory miRNA. We confirmed previously established mechanisms of miR-203a by analyzing its direct target Dlx5 in femoral head.

Our data reveal a significant effect of ovariectomy-induced bone loss, as well as the two major types of anti-osteoporotic treatment on miRNA transcription in femoral head tissue. These changes are associated with altered activity of target genes relevant to bone formation, such as Dlx5. The observed effects of bone loss and treatment response on miRNA levels in bone are also reflected in the peripheral blood, suggesting the possibility of minimally-invasive monitoring of bone-derived miRNAs using liquid biopsies.

Keywords: Bone microstructure; Circulating microRNA; Osteoporosis; Bisphosphonate; Parathyroid hormone (PTH); μCT; RUNX2

Links

DOI: 10.1016/j.bone.2019.115104

PubMed: 31683019

WoS: 000507697800003

History

Received: 2019-03-27

Revised: 2019-08-27

Accepted: 2019-10-07

Online: 2019-11-01

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Cited By (6)

Year	Entry
2021	Grillari J, Mäkitie RE, Kocijan R, Haschka J, Vázquez DC, Semmelrock E, Hackl M. Circulating miRNAs in bone health and disease. Bone. April 2021;145:115787.
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.
2021	Kerschan-Schindl K, Hackl M, Boschitsch E, Föger-Samwald U, Nägele O, Skalicky S, Weigl M, Grillari J, Pietschmann P. Diagnostic performance of a panel of miRNAs (OsteomiR) for osteoporosis in a cohort of postmenopausal women. Calcif Tiss Int. June 2021;108(6):725-737.
2020	Mäkitie RE, Hackl M, Weigl M, Frischer A, Kämpe A, Costantini A, Grillari J, Mäkitie O. Unique, gender‐dependent serum microRNA profile in PLS3 gene‐related osteoporosis. J Bone Miner Res. October 2020;35(10):1962-1973.
2021	Nevola KT, Kiel DP, Zullo AR, Weiss S, Homuth G, Foessl I, Obermayer‐Pietsch B, Motyl KJ, Lary CW. miRNA mechanisms underlying the association of beta blocker use and bone mineral density. J Bone Miner Res. January 2021;36(1):110-122.
2021	Weigl M, Kocijan R, Ferguson J, Leinfellner G, Heimel P, Feichtinger X, Pietschmann P, Grillari J, Zwerina J, Redl H, Hackl M. Longitudinal changes of circulating miRNAs during bisphosphonate and teriparatide treatment in an animal model of postmenopausal osteoporosis. J Bone Miner Res. June 2021;36(6):1131-1144.