Aurora A Kinase Inhibition Destabilizes PAX3-FOXO1 and MYCN and Synergizes with Navitoclax to Induce Rhabdomyosarcoma Cell Death

Cancer Res. 2020 Feb 15;80(4):832-842. doi: 10.1158/0008-5472.CAN-19-1479. Epub 2019 Dec 30.


The clinically aggressive alveolar rhabdomyosarcoma (RMS) subtype is characterized by expression of the oncogenic fusion protein PAX3-FOXO1, which is critical for tumorigenesis and cell survival. Here, we studied the mechanism of cell death induced by loss of PAX3-FOXO1 expression and identified a novel pharmacologic combination therapy that interferes with PAX3-FOXO1 biology at different levels. Depletion of PAX3-FOXO1 in fusion-positive (FP)-RMS cells induced intrinsic apoptosis in a NOXA-dependent manner. This was pharmacologically mimicked by the BH3 mimetic navitoclax, identified as top compound in a screen from 208 targeted compounds. In a parallel approach, and to identify drugs that alter the stability of PAX3-FOXO1 protein, the same drug library was screened and fusion protein levels were directly measured as a read-out. This revealed that inhibition of Aurora kinase A most efficiently negatively affected PAX3-FOXO1 protein levels. Interestingly, this occurred through a novel specific phosphorylation event in and binding to the fusion protein. Aurora kinase A inhibition also destabilized MYCN, which is both a functionally important oncogene and transcriptional target of PAX3-FOXO1. Combined treatment with an Aurora kinase A inhibitor and navitoclax in FP-RMS cell lines and patient-derived xenografts synergistically induced cell death and significantly slowed tumor growth. These studies identify a novel functional interaction of Aurora kinase A with both PAX3-FOXO1 and its effector MYCN, and reveal new opportunities for targeted combination treatment of FP-RMS. SIGNIFICANCE: These findings show that Aurora kinase A and Bcl-2 family proteins are potential targets for FP-RMS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aniline Compounds / pharmacology
  • Aniline Compounds / therapeutic use
  • Animals
  • Antineoplastic Combined Chemotherapy Protocols / pharmacology*
  • Antineoplastic Combined Chemotherapy Protocols / therapeutic use
  • Apoptosis / drug effects
  • Aurora Kinase A / antagonists & inhibitors
  • Aurora Kinase A / metabolism*
  • Drug Synergism
  • Gene Knockdown Techniques
  • HEK293 Cells
  • Humans
  • Mice
  • Mutagenesis, Site-Directed
  • N-Myc Proto-Oncogene Protein / metabolism*
  • Oncogene Proteins, Fusion / genetics
  • Oncogene Proteins, Fusion / metabolism*
  • Paired Box Transcription Factors / genetics
  • Paired Box Transcription Factors / metabolism*
  • Phosphorylation / drug effects
  • Primary Cell Culture
  • Protein Stability / drug effects
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • RNA, Small Interfering / metabolism
  • Rhabdomyosarcoma, Alveolar / drug therapy*
  • Rhabdomyosarcoma, Alveolar / genetics
  • Rhabdomyosarcoma, Alveolar / pathology
  • Sulfonamides / pharmacology
  • Sulfonamides / therapeutic use
  • Tumor Cells, Cultured
  • Xenograft Model Antitumor Assays


  • Aniline Compounds
  • MYCN protein, human
  • N-Myc Proto-Oncogene Protein
  • Oncogene Proteins, Fusion
  • PAX3-FOXO1A fusion protein, human
  • PMAIP1 protein, human
  • Paired Box Transcription Factors
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Small Interfering
  • Sulfonamides
  • AURKA protein, human
  • Aurora Kinase A
  • navitoclax