RESUMO
Targeting the human epidermal growth factor receptor 2 (HER2) became a landmark in the treatment of HER2-driven breast cancer. Nonetheless, the clinical efficacy of anti-HER2 therapies can be short-lived and a significant proportion of patients ultimately develop metastatic disease and die. One striking consequence of oncogenic activation of HER2 in breast cancer cells is the constitutive activation of the extracellular-regulated protein kinase 5 (ERK5) through its hyperphosphorylation. In this study, we sought to decipher the significance of this unique molecular signature in promoting therapeutic resistance to anti-HER2 agents. We found that a small-molecule inhibitor of ERK5 suppressed the phosphorylation of the retinoblastoma protein (RB) in HER2 positive breast cancer cells. As a result, ERK5 inhibition enhanced the anti-proliferative activity of single-agent anti-HER2 therapy in resistant breast cancer cell lines by causing a G1 cell cycle arrest. Moreover, ERK5 knockdown restored the anti-tumor activity of the anti-HER2 agent lapatinib in human breast cancer xenografts. Taken together, these findings support the therapeutic potential of ERK5 inhibitors to improve the clinical benefit that patients receive from targeted HER2 therapies.
Assuntos
Antineoplásicos , Neoplasias da Mama , Humanos , Feminino , Neoplasias da Mama/tratamento farmacológico , Antineoplásicos/farmacologia , Proteínas Quinases/uso terapêutico , Quinazolinas/farmacologia , Ciclo CelularRESUMO
There is overwhelming clinical evidence that the extracellular-regulated protein kinase 5 (ERK5) is significantly dysregulated in human breast cancer. However, there is no definite understanding of the requirement of ERK5 in tumor growth and metastasis due to very limited characterization of the pathway in disease models. In this study, we report that a high level of ERK5 is a predictive marker of metastatic breast cancer. Mechanistically, our in vitro data revealed that ERK5 was critical for maintaining the invasive capability of triple-negative breast cancer (TNBC) cells through focal adhesion protein kinase (FAK) activation. Specifically, we found that phosphorylation of FAK at Tyr397 was controlled by a kinase-independent function of ERK5. Accordingly, silencing ERK5 in mammary tumor grafts impaired FAK phosphorylation at Tyr397 and suppressed TNBC cell metastasis to the lung without preventing tumor growth. Collectively, these results establish a functional relationship between ERK5 and FAK signaling in promoting malignancy. Thus, targeting the oncogenic ERK5-FAK axis represents a promising therapeutic strategy for breast cancer exhibiting aggressive clinical behavior.
Assuntos
Quinase 1 de Adesão Focal/metabolismo , Proteína Quinase 7 Ativada por Mitógeno/metabolismo , Neoplasias de Mama Triplo Negativas/enzimologia , Animais , Antígenos CD/biossíntese , Antígenos CD/genética , Antígenos CD/metabolismo , Caderinas/biossíntese , Caderinas/genética , Caderinas/metabolismo , Adesão Celular/fisiologia , Linhagem Celular Tumoral , Progressão da Doença , Feminino , Xenoenxertos , Humanos , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/secundário , Camundongos , Camundongos Nus , Proteína Quinase 7 Ativada por Mitógeno/biossíntese , Proteína Quinase 7 Ativada por Mitógeno/genética , Invasividade Neoplásica , Fosforilação , Prognóstico , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/patologiaRESUMO
The extracellular signal-regulated protein kinase 5 (ERK5) is a non-redundant mitogen-activated protein kinase (MAPK) that exhibits a unique C-terminal extension which comprises distinct structural and functional properties. Here, we sought to elucidate the significance of phosphoacceptor sites in the C-terminal transactivation domain of ERK5. We have found that Thr732 acted as a functional gatekeeper residue controlling C-terminal-mediated nuclear translocation and transcriptional enhancement. Consistently, using a non-bias quantitative mass spectrometry approach, we demonstrated that phosphorylation at Thr732 conferred selectivity for binding interactions of ERK5 with proteins related to chromatin and RNA biology, whereas a number of metabolic regulators were associated with full-length wild type ERK5. Additionally, our proteomic analysis revealed that phosphorylation of the Ser730-Glu-Thr732-Pro motif could occur independently of dual phosphorylation at Thr218-Glu-Tyr220 in the activation loop. Collectively, our results firmly establish the significance of C-terminal phosphorylation in regulating ERK5 function. The post-translational modification of ERK5 on its C-terminal tail might be of particular relevance in cancer cells where ERK5 has be found to be hyperphosphoryated.
Assuntos
Proteína Quinase 7 Ativada por Mitógeno/química , Proteína Quinase 7 Ativada por Mitógeno/metabolismo , Proteômica/métodos , Treonina/metabolismo , Sítios de Ligação , Núcleo Celular/metabolismo , Células HeLa , Humanos , Espectrometria de Massas , Proteína Quinase 7 Ativada por Mitógeno/genética , Fosforilação , Ligação Proteica , Domínios Proteicos , Mapas de Interação de Proteínas , Processamento de Proteína Pós-Traducional , Transporte Proteico , Transdução de Sinais , Transcrição GênicaRESUMO
Workers are often required to perform manual materials handling tasks immediately following periods of prolonged sitting either as a secondary job component of as different tasks in a job rotation strategy. The goal of this investigation was to determine if changes to low-back kinetics and/or kinematics occurred during repetitive lifting/lowering exertions following extended seated exposures. Upper body kinematics, lumbar spine flexion angle, pelvic orientation and bilateral muscle activity from the external abdominal obliques and lumbar erector spinae were recorded for 8 males and 8 females while they alternated between sessions of repetitive lifting/lowering and prolonged sitting. Upper body kinematics were used as inputs to a linked segment model to compute low-back flexion/extension moments, compression, and shear. Peak lumbar flexion was reduced by 1.8 degrees during the lifting/lowering exertions following the first hour of sitting which consequently led to a reduction of approximately 50N in the reaction anteroposterior shear forces. Sitting postures were consistent with previously reported data. The reduced shear loads during repetitive lift/lower exertions following prolonged sitting may be a consequence of alterations in passive tissue properties which could alter the risk of low-back injury, although future research is required to examine the biomechanical significance of this finding. Changes to both kinematics and kinetics were minimal suggesting that using prolonged sitting as a component of a task series in job rotation does not alter the risk present when combined with repetitive lifting tasks.