74P - Elucidating drug resistance mechanisms using 2D and 3D culture systems

Date 15 April 2016
Event European Lung Cancer Conference 2016 (ELCC) 2016
Session Poster lunch
Topics Cancer biology
Thoracic malignancies
Translational Research
Basic Scientific Principles
Basic Principles in the Management and Treatment (of cancer)
Presenter Kenneth O'Byrne
Citation Journal of Thoracic Oncology (2016) 11 (supplement 4): S57-S166. S1556-0864(16)X0004-4
Authors K. O'Byrne1, S. Ryan2, A. Baird2, A. Urquhart2, D. Richard3, A. Davies4
  • 1Translational Research Institute, Princess Alexandra Hospital and Queensland University of Technology, QLD 4102 - Brisbane/AU
  • 2Cancer And Ageing Research Program And Translational Cell Imaging Queensland, Queensland University of Technology, Brisbane/AU
  • 3Institute Of Health And Biomedical Innovation, Cancer And Ageing Research Program, Queensland University of Technology, Brisbane/AU
  • 4Translational Cell Imaging Queensland, Queensland University of Technology, QLD 4102 - Brisbane/AU

Abstract

Background

Chemoresistance is a major obstacle to lung cancer therapy; however the key mechanisms that promote it have yet to be fully elucidated. In order to address this issue many are now resorting to three dimensional (3D) based cellular assay systems. These systems facilitate the formation of multicellular structures (MCS) whose internal microenvironment mimics closely that of those in vivo. This study compared drug resistant models of non-small cell lung cancer (NSCLC) cultured in 3D with those cultured in standard two dimensional (2D) monolayers. The behaviour of cells cultured in these distinct geometric configurations was compared by measuring viability and assessing relevant morphological changes over time.

Methods

An isogenic NSCLC cell line model of cisplatin resistance was utilised for this study. This model comprised sensitive parent (PT) and matched cisplatin resistant (CisR) lines. 3D MCS were cultured in Happy Cell ASM® for a period of several weeks. At various time points 2D and 3D cultures were treated with a range of cisplatin concentrations for 72 h. Subsequent viability assays compared the response of PT and CisR cells in 2D and 3D culture systems. Morphological analysis of the 3D models was also performed at each time point via high content analysis.

Results

Preliminary data indicates that at equivalent cisplatin concentrations, H460 3D MCS exhibit increased resistance compared with monolayers in both PT and CisR lines. Characterisation and further analysis is on-going up to a time-point of 56 days MCS culture. High Content imaging has identified a central necrotic core within these structures, which is a feature of the asymmetric growth patterns observed in vivo; that being a decrease in viable cells as you move inwards from the periphery of the MCS.

Conclusions

When subjected to cisplatin, MCS exhibited enhanced chemoresistance compared to their 2D counterparts. It has been suggested that MCS and their microenvironment are more representative of the in vivo scenario. Therefore, MCS may provide a more relevant in vitro model to determine mechanisms of chemoresistance and identify novel therapeutic targets. Thus, potentially providing a means to re-sensitise patients to therapy and ultimately identify predictive response markers to stratify patients for therapy.

Clinical trial identification

Legal entity responsible for the study

Queensland University of Technology

Funding

Queensland Health

Disclosure

All authors have declared no conflicts of interest.