An shRNA barcode screen provides insight into cancer cell vulnerability to MDM2 inhibitors

Thijn R Brummelkamp; Armida W M Fabius; Jasper Mullenders; Mandy Madiredjo; Arno Velds; Ron M Kerkhoven; René Bernards; Roderick L Beijersbergen

doi:10.1038/nchembio774

An shRNA barcode screen provides insight into cancer cell vulnerability to MDM2 inhibitors

Nat Chem Biol. 2006 Apr;2(4):202-6. doi: 10.1038/nchembio774. Epub 2006 Feb 13.

Authors

Thijn R Brummelkamp¹, Armida W M Fabius, Jasper Mullenders, Mandy Madiredjo, Arno Velds, Ron M Kerkhoven, René Bernards, Roderick L Beijersbergen

Affiliation

¹ Division of Molecular Carcinogenesis and Center for Biomedical Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.

PMID: 16474381
DOI: 10.1038/nchembio774

Abstract

The identification of the cellular targets of small molecules with anticancer activity is crucial to their further development as drug candidates. Here, we present the application of a large-scale RNA interference-based short hairpin RNA (shRNA) barcode screen to gain insight in the mechanism of action of nutlin-3 (1). Nutlin-3 is a small-molecule inhibitor of MDM2, which can activate the p53 pathway. Nutlin-3 shows strong antitumor effects in mice, with surprisingly few side effects on normal tissues. Aside from p53, we here identify 53BP1 as a critical mediator of nutlin-3-induced cytotoxicity. 53BP1 is part of a signaling network induced by DNA damage that is frequently activated in cancer but not in healthy tissues. Our results suggest that nutlin-3's tumor specificity may result from its ability to turn a cancer cell-specific property (activated DNA damage signaling) into a weakness that can be exploited therapeutically.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antineoplastic Agents / pharmacology
Blotting, Western
Cell Line, Tumor
Cells, Cultured
DNA Damage
Electronic Data Processing
Fibroblasts / metabolism
Gene Expression Regulation, Neoplastic*
Genes, p53
Genetic Techniques*
Humans
Imidazoles / chemistry
Intracellular Signaling Peptides and Proteins / chemistry
Mice
Microscopy, Fluorescence
Models, Chemical
Neoplasms / drug therapy*
Nuclear Proteins
Nucleic Acid Hybridization
Oligonucleotide Array Sequence Analysis
Phosphoproteins / chemistry
Piperazines / chemistry
Plasmids / metabolism
Proto-Oncogene Proteins c-mdm2 / antagonists & inhibitors*
Proto-Oncogene Proteins c-mdm2 / metabolism*
RNA Interference
RNA, Small Interfering / chemistry*
Signal Transduction
Tumor Suppressor Protein p53 / metabolism
Tumor Suppressor p53-Binding Protein 1

Substances

Antineoplastic Agents
Imidazoles
Intracellular Signaling Peptides and Proteins
Nuclear Proteins
Phosphoproteins
Piperazines
RNA, Small Interfering
TP53BP1 protein, human
Tumor Suppressor Protein p53
Tumor Suppressor p53-Binding Protein 1
nutlin 3
Proto-Oncogene Proteins c-mdm2

Associated data

PubChem-Substance/8139831
PubChem-Substance/8139832