, 2007 and Chen et al., 2009) and thus potential targets for anti-cancer drugs are suggested (Schoeberl et al., 2009). Because of the poor

identifiability of model parameters the reliability of the conclusions drawn from LSA remains a serious drawback. Therefore there is a need to develop theoretical approaches capable of addressing individual variability of signalling networks, and drawing valid predictions from the models with uncertain parameters. One suitable framework, offering appropriate mathematical apparatus, is global sensitivity analysis (GSA). In contrast to LSA, which estimates the effect of small variations of individual parameters on the model output in a HKI-272 manufacturer proximity to a single Pazopanib concentration Libraries solution, GSA allows exploration of the sensitivity of model outputs to the simultaneous perturbation of multiple parameters within a parameter space (Marino et al., 2008, Saltelli, 2004, Saltelli et al., 2008 and Zi et al., 2008). Recently there has been a growing recognition of the potential benefits of using GSA techniques for network model analysis (Balsa-Canto et al., 2010, Marino et al., 2008 and Rodriguez-Fernandez and Banga, 2010). Although examples of the application of GSA to biochemical network models are still rare, they have already shown promise for understanding

the effects of multi-parametric perturbations on biologically meaningful model outputs (Jia e al., 2007, Kim et al., 2010, Marino et al., 2008, Yoon and Deisboeck, 2009 and Zheng and Rundell, 2006). We propose a novel version of GSA, designed to explore the sensitivity of integrated model readouts to the perturbation of multiple model parameters within a parameter space, before and after a targeted anti-cancer drug is introduced into a network system. In our GSA implementation we place special emphasis on identifying a set of critical parameters, controlling the level of key output signals from the network, thereby providing a basis

for generating hypotheses on potential anti-cancer drug targets, biomarkers of drug resistance, and combinatorial therapies. The predictions drawn from our method are based on the analysis and comparison of global sensitivity profiles of key model readouts in the absence and presence of the drugs. We demonstrate the capabilities of our approach by mafosfamide applying it to our previously developed ErbB2/3 network model (Faratian et al., 2009b), exploring the sensitivity of its key model readout, pAkt, to simultaneous perturbation of all the model parameters in the absence and presence of the ErbB2 inhibitor pertuzumab. The GSA results, in addition to confirming our previous findings on the role of PTEN as one of the key biomarkers of resistance to anti-ErbB2 drugs, identified and allowed us to hypothesise that several additional network components (e.g. PDK1, PI3K, PP2A) significantly contribute to the control of network input–output behaviour. These components can be drug targets (e.g.