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Effect of Roflumilast, a Selective PDE4 Inhibitor, on Bone Phenotypes in ADO2 Mice

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Abstract

Autosomal Dominant Osteopetrosis type II (ADO2) is a rare bone disease of impaired osteoclastic bone resorption that usually results from heterozygous missense mutations in the chloride channel 7 (CLCN7) gene. We previously created mouse models of ADO2 (p.G213R) with one of the most common mutations (G215R) as found in humans and demonstrated that this mutation in mice phenocopies the human disease of ADO2. Previous studies have shown that roflumilast (RF), a selective phosphodiesterase 4 (PDE4) inhibitor that regulates the cAMP pathway, can increase osteoclast activity. We also observed that RF increased bone resorption in both wild-type and ADO2 heterozygous osteoclasts in vitro, suggesting it might rescue bone phenotypes in ADO2 mice. To test this hypothesis, we administered RF-treated diets (0, 20 and 100 mg/kg) to 8-week-old ADO2 mice for 6 months. We evaluated bone mineral density and bone micro-architecture using longitudinal in-vivo DXA and micro-CT at baseline, and 6-, 12-, 18-, and 24-week post-baseline time points. Additionally, we analyzed serum bone biomarkers (CTX, TRAP, and P1NP) at baseline, 12-, and 24-week post-baseline. Our findings revealed that RF treatment did not improve aBMD (whole body, femur, and spine) and trabecular BV/TV (distal femur) in ADO2 mice compared to the control group treated with a normal diet. Furthermore, we did not observe any significant changes in serum levels of bone biomarkers due to RF treatment in these mice. Overall, our results indicate that RF does not rescue the osteopetrotic bone phenotypes in ADO2 heterozygous mice.

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Funding

This work was supported by the US National Institutes of Health grant AG069583.

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Authors and Affiliations

  1. Division of Endocrinology, Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, 46202, USA

    Imranul Alam, Sara L. Hardman, Rita L. Gerard-O’Riley, Dena Acton, Reginald S. Parker & Michael J. Econs

  2. Medical and Molecular Genetics, Indiana University School of Medicine, 1120 West Michigan St, CL459, Indianapolis, IN, 46202, USA

    Michael J. Econs

  3. Biomedical Sciences and Comprehensive Care, Indiana University School of Dentistry, Indianapolis, IN, 46202, USA

    Jung Min Hong & Angela Bruzzaniti

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  1. Imranul Alam
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Contributions

Study design: IA, AB, and MJE. Study conduct: IA, SLH, RLG, DA, RSP, AB, and MJE. Data analysis: IA, SLH, RLG, AB, and MJE. Data interpretation: IA, SLH, RLG, DA, RSP, JMH, AB, and MJE. Drafting manuscript: IA, RLG, AB, and MJE. Revising manuscript content: IA, RLG, AB, and MJE. Approval of final version of manuscript: IA, SLH, RLG, DA, RSP, JMH, AB, and MJE.

Corresponding author

Correspondence to Imranul Alam.

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Conflict of interests

Imranul Alam, Sara L. Hardman, Rita L. Gerard‑O’Riley, Dena Acton, Reginald S. Parker, Jung Min Hong, Angela Bruzzaniti, and Michael J. Econs declare that they have no conflict of interest.

Human and Animal Rights and Informed Consent

All procedures involving animals performed in the study followed the guidelines of the Indiana University Animal Care and Use Committee (IACUC Protocol No. 11378). This study does not involve research in humans.

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Alam, I., Hardman, S.L., Gerard-O’Riley, R.L. et al. Effect of Roflumilast, a Selective PDE4 Inhibitor, on Bone Phenotypes in ADO2 Mice. Calcif Tissue Int 114, 419–429 (2024). https://doi.org/10.1007/s00223-023-01180-2

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