IGF-1R and mTOR Blockade: Novel Resistance Mechanisms and Synergistic Drug Combinations for Ewing Sarcoma

J Natl Cancer Inst. 2016 Aug 30;108(12):djw182. doi: 10.1093/jnci/djw182. Print 2016 Dec.

Abstract

Background: Therapies cotargeting insulin-like growth factor receptor 1 (IGF-1R) and mammalian target of rapamycin (mTOR) have demonstrated remarkable, albeit short-lived, clinical responses in a subset of Ewing sarcoma (ES) patients. However, the mechanisms of resistance and applicable strategies for overcoming drug resistance to the IGF-1R/mTOR blockade are still undefined.

Methods: To elucidate predominant mechanism(s) of acquired drug resistance while identifying synergistic drug combinations that improve clinical efficacy, we generated more than 18 ES cell lines resistant to IGF-1R- or mTOR-targeted therapy. Two small-molecule inhibitors of IGF-1R were chosen, NVP-ADW-742 (IGF-1R-selective) and OSI-906 (a dual IGF-1R/insulin receptor alpha [IR-α] inhibitor). Reverse-phase protein lysate arrays (RPPAs) revealed proteomic changes linked to IGF-1R/mTOR resistance, and selected proteins were validated in cell-based assays, xenografts, and within human clinical samples. All statistical tests were two-sided.

Results: Novel mechanisms of resistance (MOR) emerged after dalotuzumab-, NVP-ADW-742-, and OSI-906-based targeting of IGF-1R. MOR to dalotuzumab included upregulation of IRS1, PI3K, and STAT3, as well as p38 MAPK, which was also induced by OSI-906. pEIF4E(Ser209), a key regulator of Cap-dependent translation, was induced in ridaforolimus-resistant ES cell lines. Unique drug combinations targeting IGF-1R and PI3K-alpha or Mnk and mTOR were synergistic in vivo and vitro (P < .001) as assessed respectively by Mantel-Cox and isobologram testing.

Conclusions: We discovered new druggable targets expressed by chemoresistant ES cells, xenografts, and relapsed human tumors. Joint suppression of these newfound targets, in concert with IGF-1R or mTOR blockade, should improve clinical outcomes.

MeSH terms

  • Adenosine Triphosphatases / antagonists & inhibitors
  • Adenosine Triphosphatases / metabolism
  • Animals
  • Antibodies, Monoclonal / administration & dosage
  • Antibodies, Monoclonal, Humanized
  • Antineoplastic Combined Chemotherapy Protocols / pharmacology
  • Antineoplastic Combined Chemotherapy Protocols / therapeutic use*
  • Bone Neoplasms / drug therapy*
  • Bone Neoplasms / metabolism*
  • Cation Transport Proteins / antagonists & inhibitors
  • Cation Transport Proteins / metabolism
  • Cell Line, Tumor
  • Copper-Transporting ATPases
  • Drug Resistance, Neoplasm
  • Drug Synergism
  • Humans
  • Imidazoles / administration & dosage
  • Insulin Receptor Substrate Proteins / metabolism
  • Male
  • Mice
  • Mice, SCID
  • Neoplasm Transplantation
  • Nucleocytoplasmic Transport Proteins / metabolism
  • Phosphatidylinositol 3-Kinase / metabolism
  • Phosphoinositide-3 Kinase Inhibitors
  • Protein Array Analysis
  • Pyrazines / administration & dosage
  • Pyrimidines / administration & dosage
  • Pyrroles / administration & dosage
  • Receptor, IGF Type 1
  • Receptors, Somatomedin / antagonists & inhibitors*
  • STAT3 Transcription Factor / metabolism
  • Sarcoma, Ewing / drug therapy*
  • Sarcoma, Ewing / metabolism*
  • Sirolimus / administration & dosage
  • Sirolimus / analogs & derivatives
  • TOR Serine-Threonine Kinases / antagonists & inhibitors*
  • Up-Regulation
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • 3-(8-amino-1-(2-phenylquinolin-7-yl)imidazo(1,5-a)pyrazin-3-yl)-1-methylcyclobutanol
  • Antibodies, Monoclonal
  • Antibodies, Monoclonal, Humanized
  • Cation Transport Proteins
  • EIF4ENIF1 protein, human
  • IGF1R protein, human
  • IRS1 protein, human
  • Imidazoles
  • Insulin Receptor Substrate Proteins
  • Nucleocytoplasmic Transport Proteins
  • Phosphoinositide-3 Kinase Inhibitors
  • Pyrazines
  • Pyrimidines
  • Pyrroles
  • Receptors, Somatomedin
  • STAT3 Transcription Factor
  • ridaforolimus
  • dalotuzumab
  • MTOR protein, human
  • Phosphatidylinositol 3-Kinase
  • Receptor, IGF Type 1
  • TOR Serine-Threonine Kinases
  • p38 Mitogen-Activated Protein Kinases
  • Adenosine Triphosphatases
  • ATP7A protein, human
  • Copper-Transporting ATPases
  • NVP ADW742
  • Sirolimus