Cell Context Is the Third Axis of Synergy for the Combination of ATR Inhibition and Cisplatin in Ewing Sarcoma

Clin Cancer Res. 2024 Aug 15;30(16):3533-3548. doi: 10.1158/1078-0432.CCR-23-3063.

Abstract

Purpose: The importance of cellular context to the synergy of DNA damage response (DDR)-targeted agents is important for tumors with mutations in DDR pathways, but less well-established for tumors driven by oncogenic transcription factors. In this study, we exploit the widespread transcriptional dysregulation of the EWS-FLI1 transcription factor to identify an effective DDR-targeted combination therapy for Ewing sarcoma.

Experimental design: We used matrix drug screening to evaluate synergy between a DNA-PK inhibitor (M9831) or an ATR inhibitor (berzosertib) and chemotherapy. The combination of berzosertib and cisplatin was selected for broad synergy, mechanistically evaluated for Ewing sarcoma selectivity, and optimized for in vivo schedule.

Results: Berzosertib combined with cisplatin demonstrates profound synergy in multiple Ewing sarcoma cell lines at clinically achievable concentrations. The synergy is due to loss of expression of the ATR downstream target CHEK1, loss of cell-cycle check-points, and mitotic catastrophe. Consistent with the goals of the project, EWS-FLI1 drives the expression of CHEK1 and five other ATR pathway members. The loss of CHEK1 expression is not due to transcriptional repression and instead caused by degradation coupled with suppression of protein translation. The profound synergy is realized in vivo with a novel optimized schedule of this combination in subsets of Ewing sarcoma models, leading to durable complete responses in 50% of animals bearing two different Ewing sarcoma xenografts.

Conclusions: These data exploit EWS-FLI1 driven alterations in cell context to broaden the therapeutic window of berzosertib and cisplatin to establish a promising combination therapy and a novel in vivo schedule. See related commentary by Ohmura and Grünewald, p. 3358.

MeSH terms

  • Animals
  • Antineoplastic Combined Chemotherapy Protocols / pharmacology
  • Antineoplastic Combined Chemotherapy Protocols / therapeutic use
  • Ataxia Telangiectasia Mutated Proteins* / antagonists & inhibitors
  • Cell Line, Tumor
  • Checkpoint Kinase 1 / antagonists & inhibitors
  • Cisplatin* / pharmacology
  • DNA Damage / drug effects
  • Drug Synergism*
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Isoxazoles
  • Mice
  • Oncogene Proteins, Fusion / antagonists & inhibitors
  • Oncogene Proteins, Fusion / genetics
  • Proto-Oncogene Protein c-fli-1* / antagonists & inhibitors
  • Proto-Oncogene Protein c-fli-1* / genetics
  • Pyrazines
  • RNA-Binding Protein EWS* / antagonists & inhibitors
  • RNA-Binding Protein EWS* / genetics
  • Sarcoma, Ewing* / drug therapy
  • Sarcoma, Ewing* / genetics
  • Sarcoma, Ewing* / pathology
  • Xenograft Model Antitumor Assays*

Substances

  • Cisplatin
  • Ataxia Telangiectasia Mutated Proteins
  • RNA-Binding Protein EWS
  • berzosertib
  • ATR protein, human
  • EWS-FLI fusion protein
  • Proto-Oncogene Protein c-fli-1
  • Oncogene Proteins, Fusion
  • Checkpoint Kinase 1
  • Isoxazoles
  • Pyrazines