3. Overall, our data suggest that gene expression profiles can be effectively used to identify putative mode(s) of action and hazards of NP exposure, in the absence of phenotypic

data. In addition to identification of hazard, it has been suggested that gene expression profiles may be useful for quantitative assessment (e.g., establishment of reference doses) of responses related to both cancer and non-cancer endpoints (Thomas et al., 2007). Benchmark doses are generally considered more informative than the no observable adverse effect level (NOAEL) in deriving reference doses as they are based on the entire dose–response relationship (Crump et al., 1995). Because alterations

in gene expression can be initiated in the absence of biological effects (e.g., adaptive or stress response pathways effective in mitigating toxic effects), it is expected that reference doses for genomics Cobimetinib INCB024360 nmr endpoints may be too sensitive for use in HHRA. However, previous analyses of 5 chemicals (i.e., 1,4-dichlorobenzene, propylene glycol mono-t-butyl ether, 1,2,3-trichloropropane, methylene chloride and naphthalene) showed that median BMD and BMDLs for the most sensitive pathways and GO categories were highly correlated with BMD and BMDLs of cancer and non-cancer endpoints (Thomas et al., 2011 and Thomas et al., 2012). In the current study, rather than choosing the most sensitive (i.e., lowest) BMDs, we focussed on the analysis of pathways that were specific to biological outcomes observed in the mice (i.e., phenotypically anchored),

and calculated BMDs for these relevant genes and pathways. The pathway-based BMDs and BMDLs calculated here for relevant pathways were actually less sensitive (i.e., higher BMDs) than those of the observed apical Farnesyltransferase endpoints. However, the mean of the minimum BMDs and BMDLs across all the pathways that we assigned as relevant to the apical endpoints (i.e., corresponding to the most sensitive genes within the relevant pathways) were similar to those of relevant apical endpoints. Median BMDs and BMDLs for the most sensitive pathways also correlate more closely with apical endpoints even though the pathways were not necessarily relevant to these endpoints. This finding supports previous examples demonstrating a 1:1 correlation between BMDs for gene expression and apical endpoints (Thomas et al., 2011 and Thomas et al., 2012). These data indicate the potential utility of using gene expression profiles in determining acceptable exposure limits for NPs. In order to determine the specific utility of pathway derived BMDs in HHRA, it will be necessary to establish a comprehensive catalogue of pathways that are actually perturbed in the event of specific adverse effects.