State-based discovery: a multidimensional screen for small-molecule modulators of EGF signaling.
As an alternative to conventional, target-oriented drug discovery, we report a strategy that identifies compounds on the basis of the state that they induce in a signaling network. Immortalized human cells are grown in microtiter plates and treated with compounds from a small-molecule library. The target network is then activated and lysates derived from each sample are arrayed onto glass-supported nitrocellulose pads. By probing these microarrays with antibodies that report on the abundance or phosphorylation state of selected proteins, a global picture of the target network is obtained. As proof of concept, we screened 84 kinase and phosphatase inhibitors for their ability to induce different states in the ErbB signaling network. We observed functional connections between proteins that match our understanding of ErbB signaling, indicating that state-based screens can be used to define the topology of signaling networks. Additionally, compounds sort according to the multidimensional phenotypes they induce, suggesting that state-based screens may inform efforts to identify the targets of biologically active small molecules.
Sevecka
Mark
M
Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA.
MacBeath
Gavin
G
D000906:Antibodies / Q000276:immunology; D000937:Antigen-Antibody Reactions; D002460:Cell Line; D016000:Cluster Analysis; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000276:immunology*; D004815:Epidermal Growth Factor / Q000737:chemistry / Q000187:drug effects / Q000276:immunology*; D006801:Humans; D019151:Peptide Library; D040081:Protein Array Analysis; D012680:Sensitivity and Specificity; D015398:Signal Transduction / Q000276:immunology*; D013329:Structure-Activity Relationship
D000906:Antibodies; D004791:Enzyme Inhibitors; D019151:Peptide Library; D004815:Epidermal Growth Factor