Evaluation of the role of gene polymorphisms in anticancer drug efficacy using in vitro models

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1 Group members and Contact details Jacques Robert graduated in medicine from the University of Strasbourg in 1974 and received hi Research projects: The objectives of our group are: (1) to identify preclinical and clinical determinants of the activity and toxicity of anticancer drugs, at the level of gene structure (polymorphisms, mutations) and expression (DNA microarrays); (2) to identify and validate new potential targets for anticancer drug action using in particular the data generated in 1; (3) to evaluate new potential drugs and drug formulations. Our approaches are not disease-oriented but concern breast, colon, lung and head-and-neck cancers. Evaluation of the role of gene polymorphisms in anticancer drug efficacy using in vitro models We have performed a series of studies aimed at identifying significant associations between the in vitro chemosensitivity of the collection of human tumour cell lines of the National Cancer Institute (NCI) and the presence of gene polymorphisms in selected genes known to be involved in DNA repair, drug transport and metabolism, drug targeting and oncogenesis. For instance, we recently showed that a polymorphism present in the promoter of the MDM2 gene was associated with increased sensitivity of the cell lines to alkylating agents, specifically in p53 wild-type cell lines. We recently introduced in our studies a second collection of human tumour cell lines, originating from the 1 / 5

2 Japanese Foundation for Cancer Research, as a validation set to the studies performed on the NCI collection. In addition, with the Greater South-West Canceropole (Cancéropôle du Grand Sud-Ouest), we recently generated a customised SNP Chip allowing the genotyping of 384 SNP simultaneously on a single DNA, in order to extend our analyses to a larger subset of candidate SNPs. This approach has opened up new research areas for the identification of original associations between clinical features and germline polymorphisms in cancer patients, generating potential new approaches for personalised cancer therapy. Evaluation of the role of gene polymorphisms in anticancer drug efficacy in cancer patients As an extension of our in vitro approaches, we have put together collections of germline DNA samples obtained during clinical studies in order to evaluate possible associations between the presence of a polymorphism and relevant clinical features (response, survival, toxicity). For instance, we recently showed that a SNP in a DNA repair gene was significantly associated with the efficacy of oxaliplatin for colorectal cancer patients. The role of this SNP in oxaliplatin efficiency has now been confirmed in a meta-analysis. Two studies are currently ongoing as ancillary studies of large clinical trials: (1) a study on breast cancer patients treated in the neo-adjuvant setting with a drug combination containing either docetaxel or not (EORTC trial); and (2) a study of metastatic colorectal cancer patients treated with protocols containing either oxaliplatin or irinotecan (CGSO Biocolon study). Other studies are planned on breast cancer patients undergoing adjuvant hormone therapy (PHRC 2009) and on lung and head-and-neck cancer patients. The SNP chip that we have designed (see above) will be used for these studies in order to explore a large array of SNPs. Identification and validation of new targets for cancer treatment In a study supported by European grants (Eurotransbio) we identified a series of genes that are overexpressed in colon cancer tissue as compared to normal mucosa and other cancers. In collaboration with Dr. Ichas' team, we have focused our efforts on ten genes which have been experimentally validated by transfection and sirna-mediated silencing in two colorectal cancer cell lines (HT29 and HCT116). One gene product is particularly interesting, since it belongs to the MAP3K family and appears therefore as potentially "druggable". The expression of this gene will be evaluated in colorectal cancers in the Bergonié tumour bank to identify correlations between its expression and the response of colorectal cancer to EGFR antibodies. Using the NCI databases, we will identify drugs whose activity is correlated with the expression of this kinase. These drugs are potential lead compounds for the development of new anticancer 2 / 5

3 agents will be tested using in vitro and in vivo models of colorectal cancer. Evaluation of new potential drugs and drug formulations We have set up several collaborative programmes for the evaluation of new potential anticancer drugs. One of them, being carried out in collaboration with the group of Prof. Jarry at the Faculty of Pharmacy of Bordeaux, concerns the evaluation of aconitine derivatives originating from a medicinal plant growing in Kyrgyzstan. A second project concerns the evaluation of oxysterol derivatives and is undertaken in collaboration with Dr Bakalara (CNRS, Montpellier). The third concerns the evaluation of gold nanoparticles encapsulating docetaxel and targeting the folate receptor and is being developed in collaboration with Prof. Astruc (University of Bordeaux). These collaborations rely upon on our long-standing expertise in cell biology and in vitro evaluat ion of drug activity. Publications: Faur N, Araud L, Laroche-Clary A, Kanno J, Toutain J, Yamori T, Robert J, Le Morvan V. Impact of the T309G polymorphism in the MDM2 gene on the cytotoxicity of antitumor agents in cellular models with different p53 status. British Journal of Cancer 2009; 101: Charasson V, Hillaire-Buys D, Solassol I, Laurand A, Pinguet F, Le Morvan V, Robert J. Involvement of gene polymorphisms of the folate pathway enzymes in gene expression and anticancer drug cytotoxicity using the NCI-60 panel as a model. European Journal of Cancer 2009; 45: Puyo S, Le Morvan V, Robert J. Impact of EGFR gene polymorphisms on anticancer drug cytotoxicity in vitro. Molecular Diagnosis and Therapy 2008; 12: / 5

4 Bellott R, Le Morvan V, Charasson V, Laurand A, Zanger U, Klein K, Colotte M, Brouste V, Bonnet J, Robert J. Functional study of the 830C>G polymorphism of the human carboxylesterase 2 gene. Cancer Chemotherapy and Pharmacology 2008; 61: Le Morvan V, Smith D, Laurand A, Brouste V, Mathoulin-Pélissier S, Soubeyran I, Bellott R, Robert J. Determination of XPD Lys751Gln and GSTP1 Ile105Val polymorphisms in colorectal cancer patients: relationships with treatment outcome. Pharmacogenomics 2007; 8: Nief N, Le Morvan V, Robert J. Involvement of gene polymorphisms of thymidylate synthase in gene expression, protein activity and anticancer drug cytotoxicity using the NCI-60 panel. European Journal of Cancer 2007; 43: Le Morvan V, Bellott R, Moisan F, Mathoulin-Pélissier S, Bonnet J, Robert J. Relationships between genetic polymorphisms and anticancer drug cytotoxicity vis-à-vis the NCI60 panel. Pharmacogenomics 2006; 7: Moisan F, Longy M, Robert J, Le Morvan V. Identification of gene polymorphisms of human DNA topoisomerase I in the National Cancer Institute panel of human tumour cell lines. British Journal of Cancer 2006; 95: Vekris A, Meynard D, Haaz MC, Bayssas M, Bonnet J, Robert J. Determinants of the clinical activity of platinum compounds: the contribution of in silico research. 4 / 5

5 Cancer Research 2004; 64: / 5

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