title: A genome-wide RNAi screen identifies novel targets of neratinib sensitivity leading to neratinib and paclitaxel combination drug treatments.
abstract: ErbB2 is frequently activated in tumors, and influences a wide array of cellular functions, including proliferation, apoptosis, cell motility and adhesion. HKI-272 (neratinib) is a small molecule pan-kinase inhibitor of the ErbB family of receptor tyrosine kinases, and shows strong antiproliferative activity in ErbB2-overexpressing breast cancer cells. We undertook a genome-wide pooled lentiviral RNAi screen to identify synthetic lethal or enhancer (synthetic modulator screen) genes that interact with neratinib in a human breast cancer cell line (SKBR-3). These genes upon knockdown would modulate cell viability in the presence of subeffective concentrations of neratinib. We discovered a diverse set of genes whose depletion selectively impaired or enhanced the viability of SKBR-3 cells in the presence of neratinib. We observed diverse pathways including EGFR, hypoxia, cAMP, and protein ubiquitination that, when co-treated with RNAi and neratinib, resulted in arrest of cell proliferation. Examining the changes of these genes and their protein products also led to a rationale for clinically relevant drug combination treatments. Treatment of cells with either paclitaxel or cytarabine in combination with neratinib resulted in a strong antiproliferative effect. The identification of novel mediators of cellular response to neratinib and the development of potential drug combination treatments have expanded our understanding of neratinib's mode-of-action for the development of more effective therapeutic regimens. Notably, our findings support a paclitaxel and neratinib phase III clinical trial in breast cancer patients.
authors.affiliation: Systems Biology, Global Biotherapeutics, Pfizer Inc., 87 Cambridgepark Drive, Cambridge, MA 02140, USA. attila.seyhan@pfizer.com
authors.forename: Attila A
authors.initials: AA
authors.lastname: Seyhan
authors.forename: Usha
authors.initials: U
authors.lastname: Varadarajan
authors.forename: Sung
authors.initials: S
authors.lastname: Choe
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: John
authors.initials: J
authors.lastname: McGraw
authors.forename: Matthew
authors.initials: M
authors.lastname: Woods
authors.forename: Stuart
authors.initials: S
authors.lastname: Murray
authors.forename: Amy
authors.initials: A
authors.lastname: Eckert
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Terence E
authors.initials: TE
authors.lastname: Ryan
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D001943:Breast Neoplasms; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D004354:Drug Screening Assays, Antitumor; D004357:Drug Synergism; D005260:Female; D052138:Genes, Neoplasm; D055106:Genome-Wide Association Study; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D017239:Paclitaxel / Q000494:pharmacology*; D011804:Quinolines / Q000494:pharmacology*; D034622:RNA Interference; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D015398:Signal Transduction
chemical_list: D000970:Antineoplastic Agents; D017239:Paclitaxel; D011804:Quinolines; D018719:Erb-b2 Receptor Tyrosine Kinases; D011994:Recombinant Proteins; C508053:ERBB2 protein, human; C487932:neratinib