Crizotinib Synergizes with Chemotherapy in Preclinical Models of Neuroblastoma

Clin Cancer Res. 2016 Feb 15;22(4):948-60. doi: 10.1158/1078-0432.CCR-15-0379. Epub 2015 Oct 5.


Purpose: The presence of an ALK aberration correlates with inferior survival for patients with high-risk neuroblastoma. The emergence of ALK inhibitors such as crizotinib has provided novel treatment opportunities. However, certain ALK mutations result in de novo crizotinib resistance, and a phase I trial of crizotinib showed a lack of response in patients harboring those ALK mutations. Thus, understanding mechanisms of resistance and defining circumvention strategies for the clinic is critical.

Experimental design: The sensitivity of human neuroblastoma-derived cell lines, cell line-derived, and patient-derived xenograft (PDX) models with varying ALK statuses to crizotinib combined with topotecan and cyclophosphamide (topo/cyclo) was examined. Cultured cells and xenografts were evaluated for effects of these drugs on proliferation, signaling, and cell death, and assessment of synergy.

Results: In neuroblastoma murine xenografts harboring the most common ALK mutations, including those mutations associated with resistance to crizotinib (but not in those with wild-type ALK), crizotinib combined with topo/cyclo enhanced tumor responses and mouse event-free survival. Crizotinib + topo/cyclo showed synergistic cytotoxicity and higher caspase-dependent apoptosis than crizotinib or topo/cyclo alone in neuroblastoma cell lines with ALK aberrations (mutation or amplification).

Conclusions: Combining crizotinib with chemotherapeutic agents commonly used in treating newly diagnosed patients with high-risk neuroblastoma restores sensitivity in preclinical models harboring both sensitive ALK aberrations and de novo-resistant ALK mutations. These data support clinical testing of crizotinib and conventional chemotherapy with the goal of integrating ALK inhibition into multiagent therapy for ALK-aberrant neuroblastoma patients.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Anaplastic Lymphoma Kinase
  • Animals
  • Antineoplastic Combined Chemotherapy Protocols / pharmacology
  • Antineoplastic Combined Chemotherapy Protocols / therapeutic use*
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Crizotinib
  • Cyclophosphamide / administration & dosage
  • Dose-Response Relationship, Drug
  • Drug Synergism
  • Female
  • Humans
  • Inhibitory Concentration 50
  • Mice, SCID
  • Mutation
  • Neuroblastoma / drug therapy*
  • Neuroblastoma / genetics
  • Pyrazoles / administration & dosage
  • Pyridines / administration & dosage
  • Receptor Protein-Tyrosine Kinases / genetics
  • Topotecan / administration & dosage
  • Tumor Suppressor Protein p53 / genetics
  • Xenograft Model Antitumor Assays


  • Pyrazoles
  • Pyridines
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • Crizotinib
  • Topotecan
  • Cyclophosphamide
  • ALK protein, human
  • Alk protein, mouse
  • Anaplastic Lymphoma Kinase
  • Receptor Protein-Tyrosine Kinases