Activity of tyrosine kinase inhibitor Dasatinib in neuroblastoma cells in vitro and in orthotopic mouse model

Abstract

Stage 4 neuroblastoma (NB) is a devastating childhood cancer whose poor outcome has remained essentially unchanged in the last 20 years. Receptor tyrosine kinases have important roles in the control of proliferation, differentiation and apoptosis of NB cells. Thus, we tested the activity of second-generation tyrosine kinase inhibitor Dasatinib in human NB cell lines in vitro and in an orthotopic mouse model. Dasatinib inhibited cell viability with an IC 50 in the submicromolar range in 7 of 10 tested cell lines. In sensitive cells, Dasatinib reduced anchorage-independent growth and, in some instances, induced senescence and apoptosis. In HTLA-230 cells, Dasatinib treatment caused down-regulation of c-Kit and c-Src phosphorylation in conjunction with strong inhibition of Erk1/2 and Akt activity. To test the efficacy of Dasatinib in vivo, HTLA-230 and SY5Y cells were orthotopically injected in the adrenal gland of nude mice and drug treatments carried out until day 40. In mice injected with HTLA-230 cells, tumour growth was significantly inhibited at the dose of 30 mg/(kg day) when treatment was started 7 days after injection. In animals injected with SY5Y cells that were exquisitely sensitive in vitro (IC 50 5 92 nM), the antitumour effect of Dasatinib was observed at the dose of 60 mg/(kg day) but only when treatment was started 1 day after injection. However, the anti-tumour effect of Dasatinib in vivo was partial in both orthotopic models, emphasizing the importance of testing candidate new drugs in animal environments closely mimicking the human tumour. ' UICCKey words: neuroblastoma; Dasatinib; orthotopic model Neuroblastoma (NB) is the most common childhood extracranial tumour. 1 NB derives from precursor cells of the sympatoadrenal lineage and it can develop anywhere in the sympathetic system. 1 About 40% of NBs at diagnosis are localized tumours which, in general, respond to chemotherapy and have an outcome that spans from favourable (Stages 1, 2 and 3 with no MYCN amplification) to intermediate/poor (Stages 2 and 3 with MYCN amplification). 2 However, the greatest clinical challenge is represented by Stage 4 in children older than 18 months, which accounts for 50% of NB cases at diagnosis. Stage 4 NB is characterized by metastases to distant sites such as cortical bone, bone marrow, and lymph nodes non-contiguous to the primary tumour. 2 Outcome for these patients is generally very poor, a 5-year survival not exceeding 30%. 1 A striking clinical phenotype of NB is stage 4S (S5 special), which occurs in about 8% of cases. 2 These infants have a small primary tumour with widespread involvement of liver, skin, and/or bone marrow, which spontaneously regress in a substantial number of cases. Outcome of 4S tumours is similar to Stages 1 and 2 unless other unfavourable prognostic markers are present. 3 Beside tumour stage, many clinical and genetic features are utilized in NB to modulate appropriate treatment and accurately define prognosis. Age >18 months 4 and histology 5 are potent indicators of poorer ou...

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“…Chronic myeloid leukemia [124,125] Dasatinib Ovarian, Neuroblastoma [126][127][128][129] N-phenyl-4-(thiazol-5-yl)pyrimidin-2-amines Aurora kinases [130] ( …”