Screening for DAX1/EWS-FLI1 functional inhibitors identified dihydroorotate dehydrogenase as a therapeutic target for Ewing's sarcoma

Cancer Med. 2023 Apr;12(8):9802-9814. doi: 10.1002/cam4.5741. Epub 2023 Feb 24.

Abstract

Objective: EWS-FLI1 is the most common oncogenic fusion protein in Ewing's sarcoma family tumors (ESFTs). DAX1, an orphan member of the nuclear receptor superfamily, is up-regulated by EWS-FLI1 and plays a key role in the transformed phenotype of ESFTs.

Methods: To discover a functional inhibitor of DAX1 and EWS-FLI1, we screened small-molecular inhibitors using a DAX1 reporter assay system.

Results: K-234 and its derivatives, which were dihydroorotate dehydrogenase (DHODH) inhibitors, showed inhibitory effects in the reporter assay. K-234 inhibited the growth of Ewing's sarcoma with various fusion types, and K-234 derivatives altered the expression of EWS-FLI1-regulated genes. The DAX1 expression had no effect on the growth inhibitory effect of the K-234 derivatives, while DHODH overexpression or uridine treatment attenuated their inhibitory effects, suggesting that inhibition by K-234 derivatives occurs through DHODH inhibition. An in vivo study showed that a K-234 derivative clearly inhibited tumor growth in an Ewing's sarcoma xenograft mouse model.

Conclusion: Taken together, the present results suggest that DHODH inhibitors can inhibit the function of DAX1/EWS-FLI1 in ESFTs and might be a therapeutic agent with potent anti-tumor activity for Ewing's sarcoma patients.

Keywords: DAX1; EWS-FLI1; Ewing's sarcoma; dihydroorotate dehydrogenase; nucleotide pool.

MeSH terms

  • Animals
  • Dihydroorotate Dehydrogenase
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Mice
  • Oncogene Proteins, Fusion / genetics
  • Oncogene Proteins, Fusion / metabolism
  • Proto-Oncogene Protein c-fli-1 / genetics
  • RNA-Binding Protein EWS / genetics
  • Sarcoma, Ewing* / drug therapy
  • Sarcoma, Ewing* / genetics
  • Sarcoma, Ewing* / metabolism

Substances

  • EWS-FLI fusion protein
  • Dihydroorotate Dehydrogenase
  • Proto-Oncogene Protein c-fli-1
  • RNA-Binding Protein EWS
  • Oncogene Proteins, Fusion