[2015 WCI Conference] Predicting Drug-induced Skin Rash Using BioMAP Human Primary Cell-based Systems

[2015 WCI Conference] Predicting Drug-induced Skin Rash Using BioMAP Human Primary Cell-based Systems
Version:
081715

File Name/Number:
20486

Year:
2015

The relationship between drug efficacy and adverse events (AE) is dependent on (i) target selectivity, (ii) primary and secondary effects of target modulation and (iii) the concentration at which these interactions occur.  BioMAP technology uses a systems biology approach to capture the complexity of physiologically relevant tissue microenvironments and provide information about how compounds behave in these settings with respect to efficacy and safety. BioMAP systems consist of cultures of human primary cells stimulated to model disease biology and protein-based biomarker endpoints to detect phenotypic changes resulting from compound exposure. Here, we use BioMAP technology to gain a phenotypic and mechanistic understanding of the differentiating properties between classes of MAPK inhibitors. We show these inhibitors have expected anti-inflammatory and anti-proliferative activities but differ in the induction of a pro-inflammatory response associated with skin rash, an AE reported clinically for the MEK and p38 inhibitor classes.

We performed a phenotypic screen of three target-selective MAPK inhibitors: PD-184352 (MEKi), BIRB-796 (p38i), and JNK IX (JNKi). In BioMAP systems modeling inflammation, these compounds across a range of concentrations decreased several inflammatory biomarkers (CD69, IL-1a, TNFa). Interestingly, at the same exposure levels, BIRB-796 and PD-184352 showed pro-inflammatory activities (increased VCAM-1, IP-10, MIG) in a BioMAP system modeling wound healing. Multiple highly selective p38 inhibitors had similar activities, indicating that this effect is target-class, not compound specific. Conversely, JNK IX had the opposite profile in the wound healing model with anti-inflammatory activities over several concentrations.

To further elucidate the differences between the MAPK inhibitors in the wound healing system, we used phosphorylation profiling of key signaling proteins to compare underlying network interactions at early and late time points. Specifically, PD-184352, BIRB-796 and JNK IX decreased the levels of p-ERK, p-p38, and p-c-Jun, respectively, at early time points. A broader, network impact is manifested at later times suggesting crosstalk between pathways; BIRB-796 and PD-184352 both decrease p-p38 at 24 hours. Whereas PD-184352 and JNK IX decrease p-c-Jun, BIRB-796 unexpectedly increases p-c-Jun. Phosphorylation of HSP27, a cytoprotective protein implicated to have a role in skin injury, is decreased by p38i and JNKi but increased by MEKi. Together, this systems biology approach reveals that MAPK inhibitors do not act in a target-isolated manner but rather impact a network of inflammatory, proliferative and stress response pathways. In conclusion, phenotypic screening and phosphorylation profiling enables the identification of sentinel biomarkers that can be associated with efficacy and the potential for drug-related skin rash AE.