Crizotinib-induced antitumour activity in human alveolar rhabdomyosarcoma cells is not solely dependent on ALK and MET inhibition

Megiorni, Francesca; McDowell, Heather P.; Camero, Simona; Mannarino, Olga; Ceccarelli, Simona; Paiano, Milena; Losty, Paul D.; Pizer, Barry; Shukla, Rajeev; Pizzuti, Antonio; Clerico, Aldo; Dominici, Carlo

doi:10.1186/s13046-015-0228-4

Cited by 42 publications

(49 citation statements)

References 58 publications

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“…Further, the cell cycle analysis showed a G1 arrest after erlotinib (Giovannetti et al, 2008;Huether, Höpfner, Sutter, Schuppan, & Scherübl, 2005) and a G2/M arrest after crizotinib (Megiorni et al, 2015), which is in accordance with the literature. However, after the combination, the effect of crizotinib was dominant but weaker than after the monotherapy.…”

Section: Discussionsupporting

confidence: 91%

Crizotinib sensitizes the erlotinib resistant HCC827GR5 cell line by influencing lysosomal function

Steen

Keller

Dekker

et al. 2020

Journal Cellular Physiology

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In non‐small cell lung cancer, sensitizing mutations in epidermal growth factor receptor (EGFR) or cMET amplification serve as good biomarkers for targeted therapies against EGFR or cMET, respectively. Here we aimed to determine how this different genetic background would affect the interaction between the EGFR‐inhibitor erlotinib and the cMET‐inhibitor crizotinib. To unravel the mechanism of synergy we investigated the effect of the drugs on various parameters, including cell cycle arrest, migration, protein phosphorylation, kinase activity, the expression of drug efflux pumps, intracellular drug concentrations, and live‐cell microscopy. We observed additive effects in EBC‐1, H1975, and HCC827, and a strong synergism in the HCC827GR5 cell line. This cell line is a clone of the HCC827 cells that harbor an EGFR exon 19 deletion and has been made resistant to the EGFR‐inhibitor gefitinib, resulting in cMET amplification. Remarkably, the intracellular concentration of crizotinib was significantly higher in HCC827GR5 compared to the parental HCC827 cell line. Furthermore, live‐cell microscopy with a pH‐sensitive probe showed a differential reaction of the pH in the cytoplasm and the lysosomes after drug treatment in the HCC827GR5 in comparison with the HCC827 cells. This change in pH could influence the process of lysosomal sequestration of drugs. These results led us to the conclusion that lysosomal sequestration is involved in the strong synergistic reaction of the HCC827GR5 cell line to crizotinib–erlotinib combination. This finding warrants future clinical studies to evaluate whether genetic background and lysosomal sequestration could guide tailored therapeutic interventions.

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Section: Discussionsupporting

confidence: 91%

Crizotinib sensitizes the erlotinib resistant HCC827GR5 cell line by influencing lysosomal function

Steen

Keller

Dekker

et al. 2020

Journal Cellular Physiology

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“…A previous study reported that crizotinib increases intracellular ROS levels in human alveolar rhabdomyosarcoma and embryonal rhabdomyosarcoma cells [ 13 ]. Therefore, we evaluated the generation of intracellular ROS by measuring oxidized DCFH-DA (2′,7′-dichlorodihydrofluorescein-DA), and tested whether crizotinib induces apoptosis in Ba/F3 cells expressing NPM-ALK through the generation of ROS.…”

Section: Resultsmentioning

confidence: 99%

“…The mechanisms by which crizotinib exhibits its anti-tumor activity against various tumors harboring the mutation in ALK genes currently remain unclear. A recent study reported that a treatment with crizotinib resulted in the accumulation of reactive oxygen species (ROS) in human alveolar rhabdomyosarcoma and embryonal rhabdomyosarcoma cells [ 13 ], suggesting that crizotinib exhibits anti-tumor activity through the generation of ROS.…”

Section: Introductionmentioning

confidence: 99%

Alpha-tocopherol attenuates the anti-tumor activity of crizotinib against cells transformed by NPM-ALK

et al. 2017

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Anaplastic large cell lymphomas (ALCL) are mainly characterized by harboring the fusion protein nucleophosmin-anaplastic lymphoma kinase (NPM-ALK). The ALK inhibitor, crizotinib specifically induced apoptosis in Ba/F3 cells expressing NPM-ALK by inhibiting the activation of NPM-ALK and its downstream molecule, signal transducer and activator of transcription factor 3 (STAT3). We found that α-tocopherol, a major component of vitamin E, attenuated the effects of crizotinib independently of its anti-oxidant properties. Although α-tocopherol suppressed the inhibitory effects of crizotinib on the signaling axis including NPM-ALK and STAT3, it had no influence on the intake of crizotinib into cells. Crizotinib also directly inhibited the kinase activity of NPM-ALK; however, this inhibitory effect was not altered by the co-treatment with α-tocopherol. Whereas the nuclear localization of NPM-ALK was disappeared by the treatment with crizotinib, the co-treatment with α-tocopherol swept the effect of crizotinib and caused the localization of NPM-ALK in nucleus. The administration of α-tocopherol attenuated the anti-tumor activity of crizotinib against NPM-ALK-provoked tumorigenesis in vivo. Furthermore, the α-tocopherol-induced inhibition of crizotinib-caused apoptosis was also observed in NPM-ALK-positive cells derived from ALCL patients, namely, SUDHL-1 and Ki-JK. Collectively, these results not only revealed the novel mechanism underlying crizotinib-induced apoptosis in NPM-ALK-positive cells, but also suggest that the anti-tumor effects of crizotinib are attenuated when it is taken in combination with vitamin E.

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“…Due to the structural similarity of the tyrosine kinase domains across RTK families, most tyrosine kinase inhibitors lack specificity compared to monoclonal antibodies. While therefore being less suitable when studying specific RON functions, a broader target spectrum comprising RON among other, more-established RTK targets (including its interacting co-targets MET and IGF1R [6,47]) may promote the clinical development of tyrosine kinase inhibitors, such as crizotinib (NCT02612194) or ASLAN002 (also known as BMS-777607; NCT01721148) [48].…”

Section: Targeting Ron With the Imc-ron8 Antibody Strategymentioning

confidence: 99%

The Receptor Tyrosine Kinase RON and Its Isoforms as Therapeutic Targets in Ewing Sarcoma

Berning

Hennemann

Tulotta

et al. 2020

Cancers

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The receptor tyrosine kinase (RTK) RON is linked to an aggressive metastatic phenotype of carcinomas. While gaining interest as a therapeutic target, RON remains unstudied in sarcomas. In Ewing sarcoma, we identified RON among RTKs conferring resistance to insulin-like growth factor-1 receptor (IGF1R) targeting. Therefore, we explored RON in pediatric sarcoma cell lines and an embryonic Tg(kdrl:mCherry) zebrafish model, using an shRNA-based approach. To examine RON-IGF1R crosstalk, we employed the clinical-grade monoclonal antibody IMC-RON8, alone and together with the IGF1R-antibody IMC-A12. RON silencing demonstrated functions in vitro and in vivo, particularly within micrometastatic cellular capacities. Signaling studies revealed a unidirectional IGF1-mediated cross-activation of RON. Yet, IMC-A12 failed to sensitize cells to IMC-RON8, suggesting additional mechanisms of RON activation. Here, RT-PCR revealed that childhood sarcomas express short-form RON, an isoform resistant to antibody-mediated targeting. Interestingly, in contrast to carcinomas, treatment with DNA methyltransferase inhibitor did not diminish but increased short-form RON expression. Thus, this first report supports a role for RON in the metastatic progression of Ewing sarcoma. While principal molecular functions appear transferrable between carcinomas, Ewing sarcoma and possibly more common sarcoma subtypes, RON highlights that specific regulations of cellular networks and isoforms require better understanding to successfully transfer targeting strategies.

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Crizotinib-induced antitumour activity in human alveolar rhabdomyosarcoma cells is not solely dependent on ALK and MET inhibition

Cited by 42 publications

References 58 publications

Crizotinib sensitizes the erlotinib resistant HCC827GR5 cell line by influencing lysosomal function

Crizotinib sensitizes the erlotinib resistant HCC827GR5 cell line by influencing lysosomal function

Alpha-tocopherol attenuates the anti-tumor activity of crizotinib against cells transformed by NPM-ALK

The Receptor Tyrosine Kinase RON and Its Isoforms as Therapeutic Targets in Ewing Sarcoma

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