Molecular and Phenotypic Characterisation of Paediatric Glioma Cell Lines as Models for Preclinical Drug Development Dorine A. Bax1., Suzanne E. Little1., Nathalie Gaspar1,2,3, Lara Perryman1, Lynley Marshall1,2,4, Marta Viana-Pereira1,5, Tania A. Jones6, Richard D. Williams 1 , Anita Grigoriadis7, Gilles Vassal3, Paul Workman 3 , Denise Sheer6, Rui M. Reis5, Andrew D. J. Pearson1,4, Darren Hargrave4, Chris Jones1* 1 Paediatric Oncology, The Institute of Cancer Research, Sutton, United Kingdom, 2 Cancer Research UK Centre for Cancer Therapeutics, The Institute of Cancer Research, Sutton, United Kingdom, 3 Pharmacology and New Treatments of Cancer, Institut de Cance´rologie Gustave Roussy, Villejuif, France, 4 Paediatric Oncology, The Royal Marsden NHS Foundation Trust, Sutton, United Kingdom, 5 Life and Health Science Research Institute (ICVS), Universidade do Minho, Braga, Portugal, 6 Neuroscience Centre, Institute of Cell and Molecular Science, Bart’s and The London School of Medicine & Dentistry, London, United Kingdom, 7 Breakthrough Breast Cancer Unit, Guy’s Hospital, London, United Kingdom Abstract Background: Although paediatric high grade gliomas resemble their adult counterparts in many ways, there appear to be distinct clinical and biological differences. One important factor hampering the development of new targeted therapies is the relative lack of cell lines derived from childhood glioma patients, as it is unclear whether the well-established adult lines commonly used are representative of the underlying molecular genetics of childhood tumours. We have carried out a detailed molecular and phenotypic characterisation of a series of paediatric high grade glioma cell lines in comparison to routinely used adult lines. Principal Findings: All lines proliferate as adherent monolayers and express glial markers. Copy number profiling revealed complex genomes including amplification and deletions of genes known to be pivotal in core glioblastoma signalling pathways. Expression profiling identified 93 differentially expressed genes which were able to distinguish between the adult and paediatric high grade cell lines, including a number of kinases and co-ordinated sets of genes associated with DNA integrity and the immune response. Significance: These data demonstrate that glioma cell lines derived from paediatric patients show key molecular differences to those from adults, some of which are well known, whilst others may provide novel targets for evaluation in primary tumours. We thus provide the rationale and demonstrate the practicability of using paediatric glioma cell lines for preclinical and mechanistic studies. Citation: Bax DA, Little SE, Gaspar N, Perryman L, Marshall L, et al. (2009) Molecular and Phenotypic Characterisation of Paediatric Glioma Cell Lines as Models for Preclinical Drug Development. PLoS ONE 4(4): e5209. doi:10.1371/journal.pone.0005209 Editor: Mikhail V. Blagosklonny, Ordway Research Institute, United States of America Received January 13, 2009; Accepted March 19, 2009; Published April 14, 2009 Copyright: 2009 Bax et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This research is supported by Cancer Research UK (C1178/A10294, C309/A2187, C309/A8274), the Oak Foundation (LM) and La Fondation de France (NG). We acknowledge NHS funding to the NIHR Biomedical Research Centre. Paul Workman is a Cancer Research UK Life Fellow. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: chris.jones@icr.ac.uk . These authors contributed equally to this work. Introduction Paediatric high grade glioma is a clinically devastating and biologically under-studied cancer of the central nervous system. The poor clinical outcome of these children is largely due to the intrinsic drug resistance of these tumours coupled with a lack of understanding of the basic molecular pathology of the disease [1]. Unlike adults, little is known about the mechanisms of tumorigenesis of glioblastoma in children. TP53 mutation and PDGFRa overexpression, alterations associated with secondary glioblastoma in adults, have been reported at high frequencies in paediatric cases, [2], however these tumours rarely originate from pre-existing low grade lesions [3]. Furthermore, although there are clear morphological similarities between high grade gliomas of all ages, the WHO classification system is based upon adult cases, and may not be truly representative of the histopathological diversity of the childhood disease [4]. Firm conclusions are hampered by the limited number of studies performed on small sample sizes. Drug treatment efficacy also differs. In adult glioblastoma, introduction of combined chemoradiotherapy of concomitant and adjuvant temozolomide (TMZ) and radiotherapy has provided a modest survival benefit [5], particularly in patients with an epigenetically silenced O6-methylguanine-DNA-methyltransferase (MGMT) gene [6]. In children, hypermethylated MGMT promoter predicts for response to alkylating agents [7], however the survival of children treated with adjuvant temozolomide does not appear to PLoS ONE | www.plosone.org 1 April 2009 | Volume 4 | Issue 4 | e5209