MicroSCALE Screening Reveals Genetic Modifiers of Therapeutic Response in Melanoma
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Sabatini_MicroSCALE screening.pdf
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Author(s)
Wood, Kris C.
Konieczkowski, David J.
Johannessen, Cory M.
Boehm, Jesse S.
Tamayo, Pablo
Botvinnik, Olga B.
Mesirov, Jill P.
Hahn, William C.
Root, David E.
Garraway, Levi A.
Date Issued
May 2012
Journal
Science Signaling
Publisher
American Association for the Advancement of Science (AAAS)
Citation
Wood, K. C., D. J. Konieczkowski, C. M. Johannessen, J. S. Boehm, P. Tamayo, O. B. Botvinnik, J. P. Mesirov, et al. “MicroSCALE Screening Reveals Genetic Modifiers of Therapeutic Response in Melanoma.” Science Signaling 5, no. 224 (May 15, 2012): rs4-rs4.
Version
Author's final manuscript
Abstract
Cell microarrays are a promising tool for performing large-scale functional genomic screening in mammalian cells at reasonable cost, but owing to technical limitations they have been restricted for use with a narrow range of cell lines and short-term assays. Here, we describe MicroSCALE (Microarrays of Spatially Confined Adhesive Lentiviral Features), a cell microarray–based platform that enables application of this technology to a wide range of cell types and longer-term assays. We used MicroSCALE to uncover kinases that when overexpressed partially desensitized B-RAF[superscript V600E]–mutant melanoma cells to inhibitors of the mitogen-activated protein kinase kinase kinase (MAPKKK) RAF, the MAPKKs MEK1 and 2 (MEK1/2, mitogen-activated protein kinase kinase 1 and 2), mTOR (mammalian target of rapamycin), or PI3K (phosphatidylinositol 3-kinase). These screens indicated that cells treated with inhibitors acting through common mechanisms were affected by a similar profile of overexpressed proteins. In contrast, screens involving inhibitors acting through distinct mechanisms yielded unique profiles, a finding that has potential relevance for small-molecule target identification and combination drugging studies. Further, by integrating large-scale functional screening results with cancer cell line gene expression and pharmacological sensitivity data, we validated the nuclear factor κB pathway as a potential mediator of resistance to MAPK pathway inhibitors. The MicroSCALE platform described here may enable new classes of large-scale, resource-efficient screens that were not previously feasible, including those involving combinations of cell lines, perturbations, and assay outputs or those involving limited numbers of cells and limited or expensive reagents.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Whitehead Institute for Biomedical Research
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DOI of Published Version
https://doi.org/10.1126/scisignal.2002612
