RNA helicase DDX3: A novel therapeutic target in Ewing sarcoma

B. A. Wilky, C. Kim, G. McCarty, E. A. Montgomery, K. Kammers, L. R. Devine, R. N. Cole, V. Raman, David M. Loeb

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

RNA helicase DDX3 has oncogenic activity in breast and lung cancers and is required for translation of complex mRNA transcripts, including those encoding key cell-cycle regulatory proteins. We sought to determine the expression and function of DDX3 in sarcoma cells, and to investigate the antitumor activity of a novel small molecule DDX3 inhibitor, RK-33. Utilizing various sarcoma cell lines, xenografts and human tissue microarrays, we measured DDX3 expression at the mRNA and protein levels, and evaluated cytotoxicity of RK-33 in sarcoma cell lines. To study the role of DDX3 in Ewing sarcoma, we generated stable DDX3-knockdown Ewing sarcoma cell lines using DDX3-specific small hairpin RNA (shRNA), and assessed oncogenic activity. DDX3-knockdown and RK-33-treated Ewing sarcoma cells were compared with wild-type cells using an isobaric mass-tag quantitative proteomics approach to identify target proteins impacted by DDX3 inhibition. Overall, we found high expression of DDX3 in numerous human sarcoma subtypes compared with non-malignant mesenchymal cells, and knockdown of DDX3 by RNA interference inhibited oncogenic activity in Ewing sarcoma cells. Treatment with RK-33 was preferentially cytotoxic to sarcoma cells, including chemotherapy-resistant Ewing sarcoma stem cells, while sparing non-malignant cells. Sensitivity to RK-33 correlated with DDX3 protein expression. Growth of human Ewing sarcoma xenografts expressing high DDX3 was inhibited by RK-33 treatment in mice, without overt toxicity. DDX3 inhibition altered the Ewing sarcoma cellular proteome, especially proteins involved in DNA replication, mRNA translation and proteasome function. These data support further investigation of the role of DDX3 in sarcomas, advancement of RK-33 to Ewing sarcoma clinical trials and development of RNA helicase inhibition as a novel anti-neoplastic strategy.

Original languageEnglish (US)
Pages (from-to)2574-2583
Number of pages10
JournalOncogene
Volume35
Issue number20
DOIs
StatePublished - May 19 2016
Externally publishedYes

Fingerprint

RNA Helicases
Ewing's Sarcoma
Sarcoma
Protein Biosynthesis
Therapeutics
Heterografts
Cell Line
Proteins
Cell Cycle Proteins
Proteasome Endopeptidase Complex
Proteome
RNA Interference
DNA Replication
Proteomics
Small Interfering RNA
Lung Neoplasms
Stem Cells
Clinical Trials
Breast Neoplasms
Drug Therapy

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

Cite this

Wilky, B. A., Kim, C., McCarty, G., Montgomery, E. A., Kammers, K., Devine, L. R., ... Loeb, D. M. (2016). RNA helicase DDX3: A novel therapeutic target in Ewing sarcoma. Oncogene, 35(20), 2574-2583. https://doi.org/10.1038/onc.2015.336

RNA helicase DDX3 : A novel therapeutic target in Ewing sarcoma. / Wilky, B. A.; Kim, C.; McCarty, G.; Montgomery, E. A.; Kammers, K.; Devine, L. R.; Cole, R. N.; Raman, V.; Loeb, David M.

In: Oncogene, Vol. 35, No. 20, 19.05.2016, p. 2574-2583.

Research output: Contribution to journalArticle

Wilky, BA, Kim, C, McCarty, G, Montgomery, EA, Kammers, K, Devine, LR, Cole, RN, Raman, V & Loeb, DM 2016, 'RNA helicase DDX3: A novel therapeutic target in Ewing sarcoma', Oncogene, vol. 35, no. 20, pp. 2574-2583. https://doi.org/10.1038/onc.2015.336
Wilky BA, Kim C, McCarty G, Montgomery EA, Kammers K, Devine LR et al. RNA helicase DDX3: A novel therapeutic target in Ewing sarcoma. Oncogene. 2016 May 19;35(20):2574-2583. https://doi.org/10.1038/onc.2015.336
Wilky, B. A. ; Kim, C. ; McCarty, G. ; Montgomery, E. A. ; Kammers, K. ; Devine, L. R. ; Cole, R. N. ; Raman, V. ; Loeb, David M. / RNA helicase DDX3 : A novel therapeutic target in Ewing sarcoma. In: Oncogene. 2016 ; Vol. 35, No. 20. pp. 2574-2583.
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