Replacement of murine transplantation assays by 3D in vitro substitutes for the assessment of therapeutic responses of cancer stem cells.

Aims

This project aims to establish validated in vitro systems for testing the response of cancer stem cells to potential new therapeutics.

Background

Sub-populations of cancer stem cells (CSCs) have important roles in the growth and metastasis of cancers. CSCs can be resistant to chemotherapeutic agents and this is likely to have an impact on tumour recurrence. Developing effective agents is complicated by the discovery that CSC switch between an immobile epithelial phenotype and a motile epithelial-mesenchymal transformation (EMT) phenotype, with each state differing in its drug resistance pattern. Phenotypic switching is influenced by intrinsic cellular factors and the tumour microenvironment.

Tumour transplantation studies in mice are considered the 'gold standard' for characterising CSCs. Around 50 mice are used in a typical xenotransplantation study. Studies are based on the assumption that stem cell survival is too complex to be adequately modelled in vitro. However, basic CSC properties including EMT, are retained even in 2D cultures.

Previous work funded by the NC3Rs has established criteria for the development of 3D co-culture models of fibroblasts with human tumour cells including CSCs, which closely mimic cellular changes observed in vitro. This award will build on this work.

Research details and methods

To replace mouse transplantation assays for studying CSCs, a heterotypic 3D culture system, containing both tumour and stromal cells will be developed, in which in vivo cellular responses (such as invasiveness and altered therapeutic resistance) that are associated with EMT seen are mimicked. The model will be validated by comparison with the cellular heterogeneity observed in tumours and using a reporter system driven by EMT-associated gene expression changes. This will allow rapid analysis of the cellular responses to known inducers and blockers of EMT and provide the basis for a high throughput screen for assessing the efficacy of compounds to control transformation and kill CSCs in their various phenotypic manifestations.

Rodrigues MFSD et al. (2018). Effects of Cetuximab and Erlotinib on the behaviour of cancer stem cells in head and neck squamous cell carcinoma. Oncotarget 9:17(13488-13500). doi: 10.18632/oncotarget.24416

Biddle A et al. (2016). Phenotypic Plasticity Determines Cancer Stem Cell Therapeutic Resistance in Oral Squamous Cell Carcinoma. EBioMedicine 4(138-45). doi: 10.1016/j.ebiom.2016.01.007

Gammon L et al. (2016). Roles of hypoxia, stem cells and epithelial-mesenchymal transition in the spread and treatment resistance of head and neck cancer. J Oral Path Med 45(2):77-82. doi: 10.1111/jop.12327

Gemenetzidis E et al. (2015). Invasive oral cancer stem cells display resistance to ionising radiation. Oncotarget 6(41):43964077. doi: 10.18632/oncotarget.6268

Shigeishi H et al. (2015). Elevation in 5-FU-induced apoptosis in head and neck cancer stem cells by a combination of CDHP and GSK3β inhibitors. Journal of Oral Pathology & Medicine 44(3):201-7. doi: 10.1111/jop.12230

Vig N et al. (2015). Phenotypic plasticity and epithelial-to-mesenchymal transition in the behaviour and therapeutic response of oral squamous cell carcinoma. Journal of Oral Pathology & Medicine 44(9):649-55. doi: 10.1111/jop.12306

Cichoń MA et al. (2014). The receptor tyrosine kinase Axl regulates cell-cell adhesion and stemness in cutaneous squamous cell carcinoma. Oncogene 33(32):4185-92. doi: 10.1038/onc.2013.388

 

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Project grant

Status

Closed

Principal investigator

Professor Ian MacKenzie

Institution

Queen Mary University of London

Grant reference number

NC/L00061X/1

Award date

Jan 2014 - Jan 2017

Grant amount

£344,331