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Lab Invest. 2021 Dec 17; doi: 10.1038/s41374-021-00709-z. Epub 2021 Dec 17.

A murine mesenchymal stem cell model for initiating events in osteosarcomagenesis points to CDK4/CDK6 inhibition as a therapeutic target.

Laboratory investigation; a journal of technical methods and pathology

Natasja Franceschini, Raffaele Gaeta, Paul Krimpenfort, Inge Briaire-de Bruijn, Alwine B Kruisselbrink, Karoly Szuhai, Ieva Palubeckaitė, Anne-Marie Cleton-Jansen, Judith V M G Bovée

Affiliations

  1. Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands.
  2. Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Pisa, Italy.
  3. Division of Molecular Genetics, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  4. Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, The Netherlands.
  5. Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands. [email protected].

PMID: 34921235 DOI: 10.1038/s41374-021-00709-z

Abstract

Osteosarcoma is a high-grade bone-forming neoplasm, with a complex genome. Tumours frequently show chromothripsis, many deletions, translocations and copy number alterations. Alterations in the p53 or Rb pathway are the most common genetic alterations identified in osteosarcoma. Using spontaneously transformed murine mesenchymal stem cells (MSCs) which formed sarcoma after subcutaneous injection into mice, it was previously demonstrated that p53 is most often involved in the transformation towards sarcomas with complex genomics, including osteosarcoma. In the current study, not only loss of p53 but also loss of p16

© 2021. The Author(s).

References

  1. Behjati, S. et al. Recurrent mutation of IGF signalling genes and distinct patterns of genomic rearrangement in osteosarcoma. Nat. Commun. 8, 15936 (2017). - PubMed
  2. Stephens, P. J. et al. Massive genomic rearrangement acquired in a single catastrophic event during cancer development. Cell 144, 27–40 (2011). - PubMed
  3. Cortes-Ciriano, I. et al. Comprehensive analysis of chromothripsis in 2,658 human cancers using whole-genome sequencing. Nat. Genet. 52, 331–341 (2020). - PubMed
  4. Perry, J. A. et al. Complementary genomic approaches highlight the PI3K/mTOR pathway as a common vulnerability in osteosarcoma. Proc. Natl Acad. Sci. USA 111, E5564–E5573 (2014). - PubMed
  5. Kovac, M. et al. Exome sequencing of osteosarcoma reveals mutation signatures reminiscent of BRCA deficiency. Nat. Commun. 6, 8940 (2015). - PubMed
  6. Weinberg, R. The retinoblastoma protein and cell cycle control. Cell 81, 323–330 (1995). - PubMed
  7. Sayles, L. C. et al. Genome-informed targeted therapy for osteosarcoma. Cancer Discov. 9, 46–63 (2019). - PubMed
  8. Mejia-Guerrero, S., et al. Characterization of the 12q15MDM2and 12q13-14CDK4amplicons and clinical correlations in osteosarcoma. Genes Chromosom. Cancer https://doi.org/10.1002/gcc.20761 , NA-NA (2010). - PubMed
  9. Burkhart, D. L. & Sage, J. Cellular mechanisms of tumour suppression by the retinoblastoma gene. Nat. Rev. Cancer 8, 671–682 (2008). - PubMed
  10. Sherr, C. J. & Weber, J. D. The ARF/p53 pathway. Curr. Opin. Genet. Dev. 10, 94–99 (2000). - PubMed
  11. Miller, C. W. et al. Alterations of the p15, p16 and p18 Genes in Osteosarcoma. Cancer Genet. Cytogenet. 86, 136–142 (1996). - PubMed
  12. Tsuchiya, T. et al. Analysis of the p16INK4, p14ARF, p15, TP53, and MDM2 Genes and Their Prognostic Implications in Osteosarcoma and Ewing Sarcoma. Cancer Genet. Cytogenet. 120, 91–98 (2000). - PubMed
  13. Mohseny, A. B. et al. Osteosarcoma originates from mesenchymal stem cells in consequence of aneuploidization and genomic loss of Cdkn2. J. Pathol. 219, 294–305 (2009). - PubMed
  14. Righi, A. et al. p16 expression as a prognostic and predictive marker in high-grade localized osteosarcoma of the extremities: an analysis of 357 cases. Hum. Pathol. 58, 15–23 (2016). - PubMed
  15. Kosemehmetoglu, K., Ardic, F., Karslioglu, Y., Kandemir, O. & Ozcan, A. p16 expression predicts neoadjuvant tumor necrosis in osteosarcomas: reappraisal with a larger series using whole sections. Hum. Pathol. 50, 170–175 (2016). - PubMed
  16. Bu, J., et al. P16Ink4a overexpression and survival in osteosarcoma patients: a meta analysis. Int. J. Clin. Exp. Pathol. 7, 6091–6096 (2014). - PubMed
  17. Borys, D. et al. P16 expression predicts necrotic response among patients with osteosarcoma receiving neoadjuvant chemotherapy. Hum. Pathol. 43, 1948–1954 (2012). - PubMed
  18. Maitra, A., Roberts, H., Weinberg, A. & Geradts, J. Loss of p16ink4a expression correlates with decreased survival in pediatric osteosarcomas. Int. J. Cancer 95, 34–8 (2001). - PubMed
  19. Franceschini, N., et al. Transformed murine and canine mesenchymal stem cells as a model for sarcoma with complex genomics. Cancers 13, 1126 (2021). - PubMed
  20. Krimpenfort, P. et al. p15Ink4b is a critical tumour suppressor in the absence of p16Ink4a. Nature 448, 943–946 (2007). - PubMed
  21. Ottaviano, L. et al. Molecular characterization of commonly used cell lines for bone tumor research: a trans-European EuroBoNet effort. Genes Chromosom. Cancer 49, 40–51 (2010). - PubMed
  22. Zhang, W. et al. Optimization of the formation of embedded multicellular spheroids of MCF-7 cells: how to reliably produce a biomimetic 3D model. Anal. Biochem. 515, 47–54 (2016). - PubMed
  23. Hafner, M., Niepel, M., Chung, M. & Sorger, P. K. Growth rate inhibition metrics correct for confounders in measuring sensitivity to cancer drugs. Nat. Methods 13, 521–527 (2016). - PubMed
  24. WHO Classification of Tumours of Soft Tissue and Bone 5th edn, Vol. 3 (WHO Classification of Tumours Editorial Board, 2020). - PubMed
  25. Mohseny, A. B. et al. Small deletions but not methylation underlie CDKN2A/p16 loss of expression in conventional osteosarcoma. Genes Chromosom. Cancer 49, 1095–1103 (2010). - PubMed
  26. Buddingh, E. P. et al. Tumor-infiltrating macrophages are associated with metastasis suppression in high-grade osteosarcoma: a rationale for treatment with macrophage activating agents. Clin. Cancer Res. 17, 2110–2119 (2011). - PubMed
  27. Schrage, Y. M. et al. Central chondrosarcoma progression is associated with pRb pathway alterations: CDK4 down-regulation and p16 overexpression inhibit cell growth in vitro. J. Cell Mol. Med. 13, 2843–2852 (2009). - PubMed
  28. Gong, W. et al. Cyclin-dependent kinase 6 (CDK6) is a candidate diagnostic biomarker for early non-small cell lung cancer. Transl. Cancer Res. 9, 95–103 (2020). - PubMed
  29. Bankhead, P. et al. QuPath: open source software for digital pathology image analysis. Sci. Rep. 7, 16878 (2017). - PubMed
  30. Kresse, S. H. et al. Integrative analysis reveals relationships of genetic and epigenetic alterations in osteosarcoma. PLoS ONE 7, e48262 (2012). - PubMed
  31. Chow, T., et al. Creating in vitro three-dimensional tumor models: a guide for the biofabrication of a primary osteosarcoma model. Tissue Eng. Part B https://doi.org/10.1089/ten.teb.2020.0254 (2020). - PubMed
  32. De Luca, A. et al. Relevance of 3d culture systems to study osteosarcoma environment. J. Exp. Clin. Cancer Res. 37, 2 (2018). - PubMed
  33. Gao, S., Shen, J., Hornicek, F. & Duan, Z. Three-dimensional (3D) culture in sarcoma research and the clinical significance. Biofabrication 9, 032003 (2017). - PubMed
  34. Sherr, C. J., Beach, D. & Shapiro, G. I. Targeting CDK4 and CDK6: from discovery to therapy. Cancer Discov. 6, 353–367 (2016). - PubMed
  35. Guha, M. Cyclin-dependent kinase inhibitors move into Phase III. Nat. Rev. Drug Discov. 11, 892–894 (2012). - PubMed
  36. Kovac, M., et al. The early evolutionary landscape of osteosarcoma provides clues for targeted treatment strategies. J. Pathol. https://doi.org/10.1002/path.5699 (2021). - PubMed
  37. Mohseny, A. B., Hogendoorn, P. C. & Cleton-Jansen, A. M. Osteosarcoma models: from cell lines to zebrafish. Sarcoma 2012, 417271 (2012). - PubMed
  38. Tang, Y., et al. P16 protein expression as a useful predictive biomarker for neoadjuvant chemotherapy response in patients with high-grade osteosarcoma. Medicine 96, e6714 (2017). - PubMed
  39. Zhou, Y. et al. Expression and therapeutic implications of cyclin-dependent kinase 4 (CDK4) in osteosarcoma. Biochim. Biophys. Acta Mol. Basis Dis. 1864, 1573–1582 (2018). - PubMed
  40. Perez, M., Galván, S. M., García, M. P., Marín, J. J. & Carnero, A. Efficacy of CDK4 inhibition against sarcomas depends on their levels of CDK4 and p16ink4 mRNA. Oncotarget 6, 40557–74 (2015). - PubMed
  41. Gong, X. et al. Generation of multicellular tumor spheroids with microwell-based agarose scaffolds for drug testing. PLoS ONE 10, e0130348 (2015). - PubMed
  42. Cristofanilli, M. et al. Fulvestrant plus palbociclib versus fulvestrant plus placebo for treatment of hormone-receptor-positive, HER2-negative metastatic breast cancer that progressed on previous endocrine therapy (PALOMA-3): final analysis of the multicentre, double-blind, phase 3 randomised controlled trial. Lancet Oncol. 17, 425–439 (2016). - PubMed
  43. Delmore, J. E. et al. BET bromodomain inhibition as a therapeutic strategy to target c-Myc. Cell 146, 904–917 (2011). - PubMed
  44. Finn, R. S. et al. Palbociclib and letrozole in advanced breast cancer. N. Engl. J. Med. 375, 1925–1936 (2016). - PubMed
  45. Trial of Palbociclib in Second Line of Advanced Sarcomas With CDK4 Overexpression, https://clinicaltrials.gov/ct2/show/study/NCT03242382 Accessed 18-01-2021. - PubMed
  46. Abemaciclib for Bone and Soft Tissue Sarcoma With Cyclin-Dependent Kinase (CDK) Pathway Alteration, https://clinicaltrials.gov/ct2/show/NCT04040205 Accessed 18-01-2021. - PubMed
  47. Pack, L. R., Daigh, L. H., Chung, M. & Meyer, T. Clinical CDK4/6 inhibitors induce selective and immediate dissociation of p21 from cyclin D-CDK4 to inhibit CDK2. Nat. Commun. 12, 3356 (2021). - PubMed

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