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Pilot study grant

Development of human ex vivo bone-tumour niche for the study of cancer bone disease

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At a glance

Completed
Award date
November 2014 - September 2015
Grant amount
£74,131
Principal investigator
Dr James Edwards
Institute
University of Oxford

R

  • Replacement
Read the abstract
View the grant profile on GtR

Contents

Application abstract

This proposal will standardise and validate the use of a human ex vivo assay for the study of cancer-bone disease, thereby replacing mice as the first stage model for the study of cancer cell interactions within the skeleton, and providing a new system to assess novel drugs aimed at reducing cancer metastasis to and proliferation within bone, in addition to blocking tumour-induced osteolysis.
 
Uniform slices (0.5cm thick) of human bone from cores (0.5cm diameter, approx. 5 cm length) isolated from femoral heads removed during surgery will be cultured ex vivo. Human tumour cells (breast cancer, myeloma) will be seeded within each bone slice at a known density using either A) direct injection over 30 mins, followed by an overnight colonisation and transfer to experimental well plate; B) implantation as above followed by 1hr gentle agitation prior to overnight colonisation; C) immersion of bone slice in matrigel and implantation of tumour cells within bone/gel environment, followed by culture as in A. All cultures will be maintained for either 3, 7 or 10 days in aMEM (+1%BSA) as used previously and replenished every 3 days. Anti-cancer therapies previously tested within our group (bortezomib, zoledronate) will also be added to culture media in this assay and results compared to previous in vivo studies.

As with murine skeletal samples from tumour-bone disease models, bone from this assay will be assessed by microCT scanning and histomorphometric analysis to quantify bone volume and architecture, bone mineral density, tumour and bone cell distribution. In addition, media will be collected to assess the release of bone- (alkaline- and acid phosphatase) and tumour-derived factors (PTHrP, TGF-B, myeloma-specific IgG2bk) and correlated with measurements obtained using stored serum from mice from past experiments. Representative STANDARD slices from throughout the bone core will generate baseline data with SAMPLE slices cultured under experimental conditions.

Publications

  1. Rao SR et al. (2020). Modeling the Human Bone–Tumor Niche: Reducing and Replacing the Need for Animal Data. JBMRPlus 4(4):e10356. doi: 10.1002/jbm4.10356
  2. Gooding S et al. (2019). Transcriptomic profiling of the myeloma bone-lining niche reveals BMP signalling inhibition to improve bone disease. Nature Communications 10:4533. doi: 10.1038/s41467-019-12296-1
  3. Olechnowicz SWZ et al. (2019). Multiple myeloma increases nerve growth factor and other pain-related markers through interactions with the bone microenvironment. Scientific Reports 9:14189. doi: 10.1038/s41598-019-50591-5
  4. Rao S et al. (2019). Small Animal Video Tracking for Activity and Path Analysis Using a Novel Open-Source Multi-Platform Application (AnimApp). Scientific Reports (9):12343. doi: 10.1038/s41598-019-48841-7
  5. Han HS et al. (2018). Transgenic zebrafish model for quantification and visualization of tissue toxicity caused by alloying elements in newly developed biodegradable metal. Scientific Reports 8(1):13818. doi: 10.1038/s41598-018-32313-5
  6. Jun I et al. (2018). Effect of spatial arrangement and structure of hierarchically patterned fibrous scaffolds generated by a femtosecond laser on cardiomyoblast behavior. Journal of biomedical materials research. Part A 106(6):1732-1742. doi: 10.1002/jbm.a.36374
  7. Jun I et al. (2018). Electrospun Fibrous Scaffolds for Tissue Engineering: Viewpoints on Architecture and Fabrication. International Journal of Molecular Sciences 19(3). doi: 10.3390/ijms19030745
  8. Sacitharan PK et al. (2018). Spermidine restores dysregulated autophagy and polyamine synthesis in aged and osteoarthritic chondrocytes via EP300. Experimental & Molecular Medicine 50(9):123. doi: 10.1038/s12276-018-0149-3
  9. Zhao M et al. (2018). The polyphenol resveratrol promotes skeletal growth in mice through a sirtuin 1-bone morphogenic protein 2 longevity axis. British Journal of Pharmacology 175(21):4183-4192. doi: 10.1111/bph.14477
  10. Hulley P et al. (2017). Hypoxia‐inducible factor 1‐alpha does not regulate osteoclastogenesis but enhances bone resorption activity via prolyl‐4‐hydroxylase 2. The Journal of Pathology 242(3):322-333. doi: 10.1002/path.4906