A kinase inhibitor screen identifies a dual cdc7/CDK9 inhibitor to sensitise triple-negative breast cancer to EGFR-targeted therapy

Autor: Yinghui Zhang, Marcel Smid, Jichao He, Ronan P. McLaughlin, Vera E. van der Noord, John W.M. Martens, Jevin Redel, Bob van de Water, John A. Foekens
Přispěvatelé: Medical Oncology
Rok vydání: 2018
Předmět:
CDK9/Cdc7 inhibition
Cell Survival
medicine.medical_treatment
EGFR-targeted therapy
Antineoplastic Agents
Apoptosis
Cell Cycle Proteins
Triple Negative Breast Neoplasms
Protein Serine-Threonine Kinases
Lapatinib
lcsh:RC254-282
Targeted therapy
03 medical and health sciences
0302 clinical medicine
Gefitinib
SDG 3 - Good Health and Well-being
Triple-negative breast cancer
Cell Line
Tumor
medicine
Humans
Epidermal growth factor receptor
Molecular Targeted Therapy
Protein Kinase Inhibitors
Cell Proliferation
biology
Cell growth
Gene Expression Profiling
Cell Cycle
lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens
Prognosis
Cyclin-Dependent Kinase 9
3. Good health
ErbB Receptors
Drug Resistance
Neoplasm
030220 oncology & carcinogenesis
Drug resistance
Cancer research
biology.protein
Cyclin-dependent kinase 9
Female
Erlotinib
medicine.drug
Signal Transduction
Research Article
Zdroj: Breast Cancer Research : BCR
Breast Cancer Research, 21, 77
Breast Cancer Research
Breast Cancer Research, Vol 21, Iss 1, Pp 1-15 (2019)
Breast Cancer Research, 21:77. BioMed Central Ltd.
ISSN: 1465-542X
1465-5411
Popis: Background The effective treatment of triple-negative breast cancer (TNBC) remains a profound clinical challenge. Despite frequent epidermal growth factor receptor (EGFR) overexpression and reliance on downstream signalling pathways in TNBC, resistance to EGFR-tyrosine kinase inhibitors (TKIs) remains endemic. Therefore, the identification of targeted agents, which synergise with current therapeutic options, is paramount. Methods Compound-based, high-throughput, proliferation screening was used to profile the response of TNBC cell lines to EGFR-TKIs, western blotting and siRNA transfection being used to examine the effect of inhibitors on EGFR-mediated signal transduction and cellular dependence on such pathways, respectively. A kinase inhibitor combination screen was used to identify compounds that synergised with EGFR-TKIs in TNBC, utilising sulphorhodamine B (SRB) assay as read-out for proliferation. The impact of drug combinations on cell cycle arrest, apoptosis and signal transduction was assessed using flow cytometry, automated live-cell imaging and western blotting, respectively. RNA sequencing was employed to unravel transcriptomic changes elicited by this synergistic combination and to permit identification of the signalling networks most sensitive to co-inhibition. Results We demonstrate that a dual cdc7/CDK9 inhibitor, PHA-767491, synergises with multiple EGFR-TKIs (lapatinib, erlotinib and gefitinib) to overcome resistance to EGFR-targeted therapy in various TNBC cell lines. Combined inhibition of EGFR and cdc7/CDK9 resulted in reduced cell proliferation, accompanied by induction of apoptosis, G2-M cell cycle arrest, inhibition of DNA replication and abrogation of CDK9-mediated transcriptional elongation, in contrast to mono-inhibition. Moreover, high expression of cdc7 and RNA polymerase II Subunit A (POLR2A), the direct target of CDK9, is significantly correlated with poor metastasis-free survival in a cohort of breast cancer patients. RNA sequencing revealed marked downregulation of pathways governing proliferation, transcription and cell survival in TNBC cells treated with the combination of an EGFR-TKI and a dual cdc7/CDK9 inhibitor. A number of genes enriched in these downregulated pathways are associated with poor metastasis-free survival in TNBC. Conclusions Our results highlight that dual inhibition of cdc7 and CDK9 by PHA-767491 is a potential strategy for targeting TNBC resistant to EGFR-TKIs. Electronic supplementary material The online version of this article (10.1186/s13058-019-1161-9) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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