Aurora kinase B inhibition reduces the proliferation of metastatic melanoma cells and enhances the response to chemotherapy

Abstract

BackgroundThe poor response to chemotherapy and the brief response to vemurafenib in metastatic melanoma patients, make the identification of new therapeutic approaches an urgent need. Interestingly the increased expression and activity of the Aurora kinase B during melanoma progression suggests it as a promising therapeutic target.MethodsThe efficacy of the Aurora B kinase inhibitor barasertib-HQPA was evaluated in BRAF mutated cells, sensitive and made resistant to vemurafenib after chronic exposure to the drug, and in BRAF wild type cells. The drug effectiveness has been evaluated as cell growth inhibition, cell cycle progression and cell migration. In addition, cellular effectors of drug resistance and response were investigated.ResultsThe characterization of the effectors responsible for the resistance to vemurafenib evidenced the increased expression of MITF or the activation of Erk1/2 and p-38 kinases in the newly established cell lines with a phenotype resistant to vemurafenib. The sensitivity of cells to barasertib-HQPA was irrespective of BRAF mutational status. Barasertib-HQPA induced the mitotic catastrophe, ultimately causing apoptosis and necrosis of cells, inhibited cell migration and strongly affected the glycolytic metabolism of cells inducing the release of lactate. In association i) with vemurafenib the gain in effectiveness was found only in BRAF(V600K) cells while ii) with nab-paclitaxel, the combination was more effective than each drug alone in all cells.ConclusionsThese findings suggest barasertib as a new therapeutic agent and as enhancer of chemotherapy in metastatic melanoma treatment.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-015-0385-4) contains supplementary material, which is available to authorized users.

High accuracy label-free classification of kinetic cell states from holographic cytometry

“…This state was associated with targeted BRAF V600E inhibitors, apoptosis-inducing compounds, and MLN8347, consistent with the recent discoveries that AUORA kinase inhibitors are effective agents against BRAF V600E -driven melanoma cell lines (Fig. 5c) 42,43 . The morphological state associated with CDK4/6 inhibition was cyclical in normal dividing cells, but sustained in growth-arrested cells.…”

Detrimental effects of melanocortin‐1 receptor (MC1R) variants on the clinical outcomes of BRAF V600 metastatic melanoma patients treated with BRAF inhibitors

“…Considering the role of p38 in determining the apoptosis of cells, as well as its interaction with ERK (Sosa et al., ), we hypothesized that MC1R variants could act in this way by determining a low sensitivity to BRAFi. These data were also confirmed by the greater increase in vemurafenib IC50 after 21 days of exposure in Mba72 compared to Hmel1 (10 times versus 5 times), as reported in our previous work (Porcelli et al., ) and by our recent evidences in which we reported a similar behavior when cells had the same MC1R status (M3 carrying MC1R R variant and Hmel9 wildtype for MC1R). Overall, these findings could explain the shorter PFS observed in the cohort of patients with MC1R variants.…”

“…Rapidly evolving advances in the understanding of genetic and environmental influences on melanoma have been rewriting therapy, leading to major improvements in the response rate and overall survival of metastatic patients. These achievements have been obtained thanks to the discovery of drugs which act as selective inhibitors of the frequent activating mutations of BRAF kinase, as well as to the development of immunotherapies that target the checkpoint blockade CTLA4 or PD1 (Larkin et al, 2014;Long et al, 2015;Postow et al, 2015;Ribas et al, 2015). However, in this broadened and changing landscape of therapeutic options, emerging challenges are to establish a useful algorithm of sequential treatment for BRAF V600 MM patients, as well as to recognize the cellular mechanisms underlying drug response and resistance to BRAFi.…”

“…Hmel1 and Mba72 were treated as described in Porcelli et al. (Porcelli et al., ) in order to acquire resistance to vemurafenib.…”

“…The cell culture medium used was high-glucose Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS), 1% (v/v) L-glutamine, and 1% (v/v) penicillin/streptomycin, at 37°C in a humidified atmosphere at 5% CO 2 . Hmel1 and Mba72 were treated as described in Porcelli et al (Porcelli et al, 2015) in order to acquire resistance to vemurafenib. For real-time PCR, purification of total RNA from melanoma cell lines was carried out using the RNeasy Mini Kit (Qiagen), according to the manufacturer's protocol.…”