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Development and characterization of murine models of medulloblastoma extraneural growth in bone

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Abstract

Medulloblastoma is a malignant pediatric brain neoplasm with an unusual predilection for metastasis to the skeleton. The objective of this study was to generate and characterize murine models of medulloblastoma extraneural growth in bone as ‘discovery tools’ for the identification of unrecognized signal transduction pathways and factors driving metastatic bone disease. To this end, the human Daoy and D283 medulloblastoma cell lines were inoculated into the intratibial medullary space of athymic nude mice. Daoy injected mice developed a primarily osteolytic radiographic and histological phenotype. In contrast, both areas of osteolytic and osteosclerotic activity were evident in D283 inoculated bones. D283 and Daoy cell conditioned media increased in vitro osteoblast differentiation and is consistent with the enhanced bone turnover characteristic of bone metastases. Daoy cells also significantly increased bone marrow osteoclast formation, consistent with the robust in vivo osteolytic phenotype. A survey of secreted factors implicated in bone metastasis and expressed by D283 and Daoy was performed. High expression of the bone-homing factor, CXCR4, was observed in both Daoy and D283 tissues. Consistent with the skeletal phenotypes, Daoy cells, while secreting the osteoblastic factor ET-1, abundantly produced the osteolytic factors RANKL, PTHrP and TNFα. D283 cells produced high levels of both RANKL and ET-1. These newly described animal models of medulloblastoma bone metastasis are expected to serve as platforms to aid in the elucidation of novel bone metastasis signaling cascades and to test therapeutics that target both medulloblastoma metastasis and the primary tumor.

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Abbreviations

ATCC:

American Type Culture Collection

CXCR4:

C-X-C chemokine receptor 4

D283:

D283 Med

DAB:

Diaminobenzidine

DKK1:

Dickkopf homologue 1

ET-1:

Endothelin-1

HRP:

Horseradish peroxidase

PTHrP:

Parathyroid hormone-related protein

RANKL:

Receptor activator of nuclear factor kappa-β ligand

TACE:

TNF-a-converting enzyme

TGFβ:

Transforming growth factor beta

TNFα:

Tumor necrosis factor alpha

TRAP:

Tartrate-resistant acid phosphatase

WHO:

World Health Organization

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Acknowledgments

We would like to acknowledge the University of Alabama at Birmingham Center for Metabolic Bone Disease (CMBD) Histomorphometry and Animal Phenotyping Cores for their excellent services and for providing expertise on the described experiments. We would also like to extend our gratitude to Dr. Burt Nabors and Katrina Clines for their assistance on this project. This study was supported by NIH K08 CA118428 (G.C.) and the University of Alabama at Birmingham Center for Metabolic Bone Disease (CMBD) NIH T32AR047512 and P30AR046031.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Division of Endocrinology, Diabetes and Metabolism, Department of Medicine, University of Alabama at Birmingham, Boshell Diabetes Building RM 730B, 1808 7th Avenue South, Birmingham, AL, 35294-0012, USA

    Jessica M. Grunda & Gregory A. Clines

  2. Division of Orthopaedic Surgery, Department of Surgery, University of Alabama at Birmingham, Birmingham, AL, USA

    Dezhi Wang

  3. Veterans Affairs Medical Center, Birmingham, AL, USA

    Gregory A. Clines

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  1. Jessica M. Grunda
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  2. Dezhi Wang
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  3. Gregory A. Clines
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Correspondence to Gregory A. Clines.

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Grunda, J.M., Wang, D. & Clines, G.A. Development and characterization of murine models of medulloblastoma extraneural growth in bone. Clin Exp Metastasis 30, 769–779 (2013). https://doi.org/10.1007/s10585-013-9577-6

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