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Dkk1 KO Mice Treated with Sclerostin Antibody Have Additional Increases in Bone Volume

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Abstract

Dickkopf-1 (DKK1) and sclerostin are antagonists of the Wnt/β-catenin pathway and decreased expression of either results in increased bone formation and mass. As both affect the same signaling pathway, we aimed to elucidate the redundancy and/or compensation of sclerostin and DKK1. Weekly sclerostin antibody (Scl-Ab) was used to treat 9-week-old female Dkk1 KO (Dkk1−/−:Wnt3+/−) mice and compared to Scl-Ab-treated wild-type mice as well as vehicle-treated Dkk1 KO and wild-type animals. While Wnt3 heterozygote (Wnt3+/−) mice show no bone phenotype, Scl-Ab and vehicle-treated control groups of this genotype were included. Specimens were harvested after 3 weeks for microCT, bone histomorphometry, anti-sclerostin immunohistochemistry, and biomechanical testing. Scl-Ab enhanced bone anabolism in all treatment groups, but with synergistic enhancement seen in the cancellous compartment of Dkk1 KO mice (bone volume + 55% Dkk1 KO p < 0.01; + 22% wild type p < 0.05). Scl-Ab treatment produced less marked increases in cortical bone of the tibiae, with anabolic effects similar across genotypes. Mechanical testing confirmed that Scl-Ab improved strength across all genotypes; however, no enhancement was seen within Dkk1 KO mice. Dynamic bone labeling showed that Scl-Ab treatment was associated with increased bone formation, regardless of genotype. Immunohistochemical staining for sclerostin protein indicated no differences in the Dkk1 KO mice, indicating that the increased Wnt signaling associated with DKK1 deficiency was not compensated by upregulation of sclerostin protein. These data suggest complex interactions between Wnt signaling factors in bone, but critically illustrate synergy between DKK1 deficiency and Scl-Ab treatment. These data support the application of dual-targeted therapeutics in the modulation of bone anabolism.

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Authors and Affiliations

  1. Orthopaedic Research & Biotechnology Unit, The Children’s Hospital at Westmead, Locked Bag 4001, Westmead, NSW, 2145, Australia

    Alyson Morse, Tegan L. Cheng, Aaron Schindeler & David G. Little

  2. Discipline of Paediatrics and Child Health, Faculty of Medicine, University of Sydney, Sydney, Australia

    Alyson Morse, Tegan L. Cheng, Aaron Schindeler & David G. Little

  3. Bone Biology Program, The Garvan Institute of Medical Research, Sydney, Australia

    Michelle M. McDonald & Sindhu T. Mohanty

  4. Novartis Pharma AG, Basel, Switzerland

    Michaela Kneissel & Ina Kramer

Authors
  1. Alyson Morse
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  2. Tegan L. Cheng
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  3. Aaron Schindeler
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  4. Michelle M. McDonald
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  5. Sindhu T. Mohanty
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  6. Michaela Kneissel
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  7. Ina Kramer
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  8. David G. Little
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Contributions

Study design: AM, DGL, MMM, MK, IK. Study conduct: AM, TLC, SM. Data collection: AM, TLC. Data interpretation: AM, TLC, DGL. Drafting manuscript: AM. Revising manuscript content and approving final version of manuscript: all authors. AM takes responsibility for the integrity of the data analysis. We thank Lauren Peacock and Kathy Mikulec for their assistance in animal dosing. We thank Dr. Ciara Murphy for her assistance in developing methods for confocal imaging.

Corresponding author

Correspondence to David G. Little.

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Conflict of interest

The authors received materials support (Scl-Ab) for this study from Novartis Pharma AG (Basel, Switzerland) and Mereo BioPharma (London, UK). Prof Little has received additional funding and materials support from Novartis Pharma AG and Mereo BioPharma for pre-clinical research separate to this submission and is a consultant for Orthopediatrics. Prof Little and A/Prof Schindeler have received funding support from Amgen Inc., UCB Pharma, N8 Medical, and Celgene for pre-clinical research separate to this submission. Dr McDonald received salary support from the IBMS Greg Mundy Fellowship and receives a Practitioner Fellowship (National Health & Medical Research Council). Authors Dr Kneissel and Dr Kramer are employees of Novartis Pharma AG.

Human and Animal Rights and Informed Consent

Animal experiments were approved and performed under the Western Sydney LHD Animal Ethics Committee protocol 4174 and CMRI/CHW animal ethics protocol K338.

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Morse, A., Cheng, T.L., Schindeler, A. et al. Dkk1 KO Mice Treated with Sclerostin Antibody Have Additional Increases in Bone Volume. Calcif Tissue Int 103, 298–310 (2018). https://doi.org/10.1007/s00223-018-0420-6

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  • DOI: https://doi.org/10.1007/s00223-018-0420-6

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