Recapitulation of tumor heterogeneity and molecular signatures in a 3D brain cancer model with decreased sensitivity to histone deacetylase inhibition
INTRODUCTION Physiologically relevant pre-clinical ex vivo models recapitulating CNS tumor micro-environmental complexity will aid development of biologically-targeted agents. We present comprehensive characterization of tumor aggregates generated using the 3D Rotary Cell Culture System (RCCS)....
| Main Authors: | , , , , , , , , |
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| Format: | Article |
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Public Library of Science
2012
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| Online Access: | https://eprints.nottingham.ac.uk/37648/ |
| _version_ | 1848795504297639936 |
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| author | Smith, Stuart J. Wilson, Martin Ward, Jennifer H. Rahman, Cheryl V. Peet, Andrew C. Macarthur, Donald C. Rose, Felicity R.A.J. Grundy, Richard G. Rahman, Ruman |
| author_facet | Smith, Stuart J. Wilson, Martin Ward, Jennifer H. Rahman, Cheryl V. Peet, Andrew C. Macarthur, Donald C. Rose, Felicity R.A.J. Grundy, Richard G. Rahman, Ruman |
| author_sort | Smith, Stuart J. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | INTRODUCTION
Physiologically relevant pre-clinical ex vivo models recapitulating CNS tumor micro-environmental complexity will aid development of biologically-targeted agents. We present comprehensive characterization of tumor aggregates generated using the 3D Rotary Cell Culture System (RCCS).
METHODS
CNS cancer cell lines were grown in conventional 2D cultures and the RCCS and comparison with a cohort of 53 pediatric high grade gliomas conducted by genome wide gene expression and microRNA arrays, coupled with immunohistochemistry, ex vivo magnetic resonance spectroscopy and drug sensitivity evaluation using the histone deacetylase inhibitor, Vorinostat.
RESULTS
Macroscopic RCCS aggregates recapitulated the heterogeneous morphology of brain tumors with a distinct proliferating rim, necrotic core and oxygen tension gradient. Gene expression and microRNA analyses revealed significant differences with 3D expression intermediate to 2D cultures and primary brain tumors. Metabolic profiling revealed differential profiles, with an increase in tumor specific metabolites in 3D. To evaluate the potential of the RCCS as a drug testing tool, we determined the efficacy of Vorinostat against aggregates of U87 and KNS42 glioblastoma cells. Both lines demonstrated markedly reduced sensitivity when assaying in 3D culture conditions compared to classical 2D drug screen approaches.
CONCLUSIONS
Our comprehensive characterization demonstrates that 3D RCCS culture of high grade brain tumor cells has profound effects on the genetic, epigenetic and metabolic profiles of cultured cells, with these cells residing as an intermediate phenotype between that of 2D cultures and primary tumors. There is a discrepancy between 2D culture and tumor molecular profiles, and RCCS partially re-capitulates tissue specific features, allowing drug testing in a more relevant ex vivo system. |
| first_indexed | 2025-11-14T19:33:08Z |
| format | Article |
| id | nottingham-37648 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:33:08Z |
| publishDate | 2012 |
| publisher | Public Library of Science |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-376482020-05-04T16:34:59Z https://eprints.nottingham.ac.uk/37648/ Recapitulation of tumor heterogeneity and molecular signatures in a 3D brain cancer model with decreased sensitivity to histone deacetylase inhibition Smith, Stuart J. Wilson, Martin Ward, Jennifer H. Rahman, Cheryl V. Peet, Andrew C. Macarthur, Donald C. Rose, Felicity R.A.J. Grundy, Richard G. Rahman, Ruman INTRODUCTION Physiologically relevant pre-clinical ex vivo models recapitulating CNS tumor micro-environmental complexity will aid development of biologically-targeted agents. We present comprehensive characterization of tumor aggregates generated using the 3D Rotary Cell Culture System (RCCS). METHODS CNS cancer cell lines were grown in conventional 2D cultures and the RCCS and comparison with a cohort of 53 pediatric high grade gliomas conducted by genome wide gene expression and microRNA arrays, coupled with immunohistochemistry, ex vivo magnetic resonance spectroscopy and drug sensitivity evaluation using the histone deacetylase inhibitor, Vorinostat. RESULTS Macroscopic RCCS aggregates recapitulated the heterogeneous morphology of brain tumors with a distinct proliferating rim, necrotic core and oxygen tension gradient. Gene expression and microRNA analyses revealed significant differences with 3D expression intermediate to 2D cultures and primary brain tumors. Metabolic profiling revealed differential profiles, with an increase in tumor specific metabolites in 3D. To evaluate the potential of the RCCS as a drug testing tool, we determined the efficacy of Vorinostat against aggregates of U87 and KNS42 glioblastoma cells. Both lines demonstrated markedly reduced sensitivity when assaying in 3D culture conditions compared to classical 2D drug screen approaches. CONCLUSIONS Our comprehensive characterization demonstrates that 3D RCCS culture of high grade brain tumor cells has profound effects on the genetic, epigenetic and metabolic profiles of cultured cells, with these cells residing as an intermediate phenotype between that of 2D cultures and primary tumors. There is a discrepancy between 2D culture and tumor molecular profiles, and RCCS partially re-capitulates tissue specific features, allowing drug testing in a more relevant ex vivo system. Public Library of Science 2012-12-18 Article PeerReviewed Smith, Stuart J., Wilson, Martin, Ward, Jennifer H., Rahman, Cheryl V., Peet, Andrew C., Macarthur, Donald C., Rose, Felicity R.A.J., Grundy, Richard G. and Rahman, Ruman (2012) Recapitulation of tumor heterogeneity and molecular signatures in a 3D brain cancer model with decreased sensitivity to histone deacetylase inhibition. PloS one, 7 (12). e52335. ISSN 1932-6203 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0052335 doi:10.1371/journal.pone.0052335 doi:10.1371/journal.pone.0052335 |
| spellingShingle | Smith, Stuart J. Wilson, Martin Ward, Jennifer H. Rahman, Cheryl V. Peet, Andrew C. Macarthur, Donald C. Rose, Felicity R.A.J. Grundy, Richard G. Rahman, Ruman Recapitulation of tumor heterogeneity and molecular signatures in a 3D brain cancer model with decreased sensitivity to histone deacetylase inhibition |
| title | Recapitulation of tumor heterogeneity and molecular signatures in a 3D brain cancer model with decreased sensitivity to histone deacetylase inhibition |
| title_full | Recapitulation of tumor heterogeneity and molecular signatures in a 3D brain cancer model with decreased sensitivity to histone deacetylase inhibition |
| title_fullStr | Recapitulation of tumor heterogeneity and molecular signatures in a 3D brain cancer model with decreased sensitivity to histone deacetylase inhibition |
| title_full_unstemmed | Recapitulation of tumor heterogeneity and molecular signatures in a 3D brain cancer model with decreased sensitivity to histone deacetylase inhibition |
| title_short | Recapitulation of tumor heterogeneity and molecular signatures in a 3D brain cancer model with decreased sensitivity to histone deacetylase inhibition |
| title_sort | recapitulation of tumor heterogeneity and molecular signatures in a 3d brain cancer model with decreased sensitivity to histone deacetylase inhibition |
| url | https://eprints.nottingham.ac.uk/37648/ https://eprints.nottingham.ac.uk/37648/ https://eprints.nottingham.ac.uk/37648/ |