shRNA kinome screen identifies TBK1 as a therapeutic target for HER2+ breast cancer.
HER2(+) breast cancer is currently treated with chemotherapy plus anti-HER2 inhibitors. Many patients do not respond or relapse with aggressive metastatic disease. Therefore, there is an urgent need for new therapeutics that can target HER2(+) breast cancer and potentiate the effect of anti-HER2 inhibitors, in particular those that can target tumor-initiating cells (TIC). Here, we show that MMTV-Her2/Neu mammary tumor cells cultured as nonadherent spheres or as adherent monolayer cells select for stabilizing mutations in p53 that "immortalize" the cultures and that, after serial passages, sphere conditions maintain TICs, whereas monolayer cells gradually lose these tumorigenic cells. Using tumorsphere formation as surrogate for TICs, we screened p53-mutant Her2/Neu(+) tumorsphere versus monolayer cells with a lentivirus short hairpin RNA kinome library. We identified kinases such as the mitogen-activated protein kinase and the TGFβR protein family, previously implicated in HER2(+) breast cancer, as well as autophagy factor ATG1/ULK1 and the noncanonical IκB kinase (IKK), TANK-binding kinase 1 (TBK1), which have not been previously linked to HER2(+) breast cancer. Knockdown of TBK1 or pharmacologic inhibition of TBK1 and the related protein, IKKε, suppressed growth of both mouse and human HER2(+) breast cancer cells. TBK1/IKKε inhibition promoted cellular senescence by suppressing p65-NF-κB and inducing p16(Ink4a). In addition, TBK1/IKKε inhibition cooperated with lapatinib, a HER2/EGFR1-targeted drug, to accelerate apoptosis and kill HER2(+) breast cancer cells both in culture and in xenografts. Our results suggest that patients with HER2(+) breast cancer may benefit from anti-TBK1/IKKε plus anti-HER2 combination therapies and establish conditions that can be used to screen for additional TIC-specific inhibitors of HER2(+) breast cancer.
Authors' Affiliations: Division of Advanced Diagnostics, Toronto General Research Institute-University Health Network; Medicinal Chemistry Platform, Ontario Institute for Cancer Research; The Donnelly Centre, University of Toronto; Program in Developmental and Stem Cell Biology, Department of Molecular Genetics, The Hospital for Sick Children; Ontario Cancer Institute, University of Toronto, and Drug Discovery Program, Department of Pharmacology and Toxicology; Toronto, Ontario, Canada.
Tao
T
Deng
Jeff C
JC
Liu
Philip E D
PE
Chung
David
D
Uehling
Ahmed
A
Aman
Babu
B
Joseph
Troy
T
Ketela
Zhe
Z
Jiang
Nathan F
NF
Schachter
Robert
R
Rottapel
Sean E
SE
Egan
Rima
R
Al-Awar
Jason
J
Moffat
Eldad
E
Zacksenhaus
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001343:Autophagy; D000071189:Autophagy-Related Protein-1 Homolog; D001943:Breast Neoplasms / Q000737:chemistry / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D005260:Female; D016158:Genes, p53; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors; D000077341:Lapatinib; D051379:Mice; D016513:Mice, SCID; D009154:Mutation; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D011799:Quinazolines / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis*; D051996:Transcription Factor RelA / Q000037:antagonists & inhibitors
D000071189:Autophagy-Related Protein-1 Homolog; D047908:Intracellular Signaling Peptides and Proteins; D000077341:Lapatinib; D017346:Protein Serine-Threonine Kinases; D011799:Quinazolines; D034741:RNA, Small Interfering; D018719:Erb-b2 Receptor Tyrosine Kinases; D051996:Transcription Factor RelA; C508053:ERBB2 protein, human; C403191:TBK1 protein, human; C484754:ULK1 protein, human