Targeting Anaplastic Lymphoma Kinase (ALK) in Rhabdomyosarcoma (RMS) with the Second-Generation ALK Inhibitor Ceritinib

Abstract

BackgroundThe receptor tyrosine kinase (RTK) anaplastic lymphoma kinase (ALK) has been implicated in the tumorigenesis of rhabdomyosarcoma (RMS). However, the exact role of ALK in RMS is debatable and remains to be elucidated.ObjectiveTo determine the in vitro and in vivo effects and mechanism of action of the second-generation ALK inhibitor ceritinib on RMS cell growth.MethodsEffects of ceritinib on cell proliferation, wound healing, cell cycle, and RTK signaling were determined in alveolar and embryonal rhabdomyosarcoma (ARMS, ERMS). In addition, possible synergistic effects of combined treatment with ceritinib and the Abl/Src family kinase inhibitor dasatinib were determined.ResultsCeritinib treatment led to decreased cell proliferation, cell cycle arrest, apoptosis, and decreased in vivo tumor growth for the ARMS subtype. ERMS cell lines were less affected and showed no cell cycle arrest or apoptosis. Both subtypes lacked intrinsic ALK phosphorylation, and ceritinib was shown to affect the IGF1R signaling pathway. High levels of phosphorylated Src (Tyr416) were present following ceritinib treatment, making combined treatment with a Src inhibitor a potential treatment option. Combined treatment of ceritinib and dasatinib showed synergistic effects in both ERMS and ARMS cell lines.ConclusionThis study shows that monotherapy with an ALK inhibitor, such as ceritinib, in RMS, has no effect on ALK signaling. However, the synergistic effects of ceritinib and dasatinib are promising, most probably due to targeting of IGF1R and Src. Electronic supplementary materialThe online version of this article (10.1007/s11523-017-0528-z) contains supplementary material, which is available to authorized users.

Olaparib and temozolomide in desmoplastic small round cell tumors: a promising combination in vitro and in vivo

“…IC 50 values were calculated using GraphPad Prism Version 5.03 software. Effects of treatment on cell migration were assessed by wound healing assays as previously described (van Erp et al 2017). Cell migration is depicted in relative gap size: gap size at t N /gap size at t 0 (t N = hours of treatment, t 0 = start of treatment).…”

“…Drug synergy of combined olaparib and TMZ treatment was assessed as previously described (van Erp et al 2017). All drug concentrations were simultaneously combined in a non-constant ratio, and the combination index (CI) and dose reduction index (DRI) were calculated using CompuSyn software.…”

“…Western Blot analysis was performed as previously described (van Erp et al 2017). Monoclonal rabbit anti-PARP1 (PARP1, 1:2000; cat.…”

Safety and Activity of the Combination of Ceritinib and Dasatinib in Osteosarcoma

“…The currently available IGF1R-targeted therapies have shown marginal efficacy in advanced clinical trials for OS and other tumor entities, presumably due to the activation of bypass signaling as a resistance mechanism. Previous reports have suggested that proto-oncogene tyrosine-protein kinase Src (Src) can mediate resistance to IGF1R inhibition in rabdomyosarcoma [12] and targeting Src with dasatinib sensitizes NSCLC cells to IGF1R tyrosine kinase inhibitors (TKIs) [13]. Src is phosphorylated in OS cells [14] and inhibition of Src activity by dasatinib induced apoptosis in OS cell lines and inhibited cell motility and invasiveness in vitro [15].…”

“…Src is phosphorylated in OS cells [14] and inhibition of Src activity by dasatinib induced apoptosis in OS cell lines and inhibited cell motility and invasiveness in vitro [15]. A synergistic effect of dasatinib (as Src inhibitor) and ceritinib (as IGF1R inhibitor) has been described in rabdomyosarcoma cell lines [12] and one phase I/II trial is recruiting rabdomyosarcoma patients to investigate the combined effects of the IGF1R antibody ganitumab and dasatinib (NCT03041701). However, the combination of ceritinib and dasatinib has not been tested in clinical studies so far.…”