Mechanism of action of trabectedin in desmoplastic small round cell tumor cells

S. Uboldi, I. Craparotta, G. Colella, E. Ronchetti, L. Beltrame, S. Vicario, S. Marchini, N. Panini, G. Dagrada, F. Bozzi, S. Pilotti, C. M. Galmarini, M. D'Incalci, R. Gatta

Research output: Contribution to journalArticle

Abstract

Background: Desmoplastic small round cell tumor (DSRCT) is a rare and highly aggressive disease, that can be described as a member of the family of small round blue cell tumors. The molecular diagnostic marker is the t(11;22)(p13;q12) translocation, which creates an aberrant transcription factor, EWS-WT1, that underlies the oncogenesis of DSRCT. Current treatments are not very effective so new active drugs are needed. Trabectedin, now used as a single agent for the treatment of soft tissue sarcoma, was reported to be active in some pre-treated DSRCT patients. Using JN-DSRCT-1, a cell line derived from DSRCT expressing the EWS-WT1 fusion protein, we investigated the ability of trabectedin to modify the function of the chimeric protein, as in other sarcomas expressing fusion proteins. After detailed characterization of the EWS-WT1 transcripts structure, we investigated the mode of action of trabectedin, looking at the expression and function of the oncogenic chimera. Methods: We characterized JN-DSRCT-1 cells using cellular approaches (FISH, Clonogenicity assay) and molecular approaches (Sanger sequencing, ChIP, GEP). Results: JN-DSRCT-1 cells were sensitive to trabectedin at nanomolar concentrations. The cell line expresses different variants of EWS-WT1, some already identified in patients. EWS-WT1 mRNA expression was affected by trabectedin and chimeric protein binding on its target gene promoters was reduced. Expression profiling indicated that trabectedin affects the expression of genes involved in cell proliferation and apoptosis. Conclusions: The JN-DSRCT-1 cell line, in vitro, is sensitive to trabectedin: after drug exposure, EWS-WT1 chimera expression decreases as well as binding on its target promoters. Probably the heterogeneity of chimera transcripts is an obstacle to precisely defining the molecular mode of action of drugs, calling for further cellular models of DSRCT, possibly growing in vivo too, to mimic the biological complexity of this disease.

Original languageEnglish
Article number107
JournalBMC Cancer
Volume17
Issue number1
DOIs
Publication statusPublished - Feb 6 2017

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trabectedin
Desmoplastic Small Round Cell Tumor
Sarcoma
Pharmaceutical Preparations
Cell Line
Proteins
Aptitude
Molecular Pathology

Keywords

  • DSRCT
  • JN-DSRCT-1
  • Trabectedin

ASJC Scopus subject areas

  • Oncology
  • Genetics
  • Cancer Research

Cite this

Mechanism of action of trabectedin in desmoplastic small round cell tumor cells. / Uboldi, S.; Craparotta, I.; Colella, G.; Ronchetti, E.; Beltrame, L.; Vicario, S.; Marchini, S.; Panini, N.; Dagrada, G.; Bozzi, F.; Pilotti, S.; Galmarini, C. M.; D'Incalci, M.; Gatta, R.

In: BMC Cancer, Vol. 17, No. 1, 107, 06.02.2017.

Research output: Contribution to journalArticle

Uboldi, S, Craparotta, I, Colella, G, Ronchetti, E, Beltrame, L, Vicario, S, Marchini, S, Panini, N, Dagrada, G, Bozzi, F, Pilotti, S, Galmarini, CM, D'Incalci, M & Gatta, R 2017, 'Mechanism of action of trabectedin in desmoplastic small round cell tumor cells', BMC Cancer, vol. 17, no. 1, 107. https://doi.org/10.1186/s12885-017-3091-1
Uboldi, S. ; Craparotta, I. ; Colella, G. ; Ronchetti, E. ; Beltrame, L. ; Vicario, S. ; Marchini, S. ; Panini, N. ; Dagrada, G. ; Bozzi, F. ; Pilotti, S. ; Galmarini, C. M. ; D'Incalci, M. ; Gatta, R. / Mechanism of action of trabectedin in desmoplastic small round cell tumor cells. In: BMC Cancer. 2017 ; Vol. 17, No. 1.
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abstract = "Background: Desmoplastic small round cell tumor (DSRCT) is a rare and highly aggressive disease, that can be described as a member of the family of small round blue cell tumors. The molecular diagnostic marker is the t(11;22)(p13;q12) translocation, which creates an aberrant transcription factor, EWS-WT1, that underlies the oncogenesis of DSRCT. Current treatments are not very effective so new active drugs are needed. Trabectedin, now used as a single agent for the treatment of soft tissue sarcoma, was reported to be active in some pre-treated DSRCT patients. Using JN-DSRCT-1, a cell line derived from DSRCT expressing the EWS-WT1 fusion protein, we investigated the ability of trabectedin to modify the function of the chimeric protein, as in other sarcomas expressing fusion proteins. After detailed characterization of the EWS-WT1 transcripts structure, we investigated the mode of action of trabectedin, looking at the expression and function of the oncogenic chimera. Methods: We characterized JN-DSRCT-1 cells using cellular approaches (FISH, Clonogenicity assay) and molecular approaches (Sanger sequencing, ChIP, GEP). Results: JN-DSRCT-1 cells were sensitive to trabectedin at nanomolar concentrations. The cell line expresses different variants of EWS-WT1, some already identified in patients. EWS-WT1 mRNA expression was affected by trabectedin and chimeric protein binding on its target gene promoters was reduced. Expression profiling indicated that trabectedin affects the expression of genes involved in cell proliferation and apoptosis. Conclusions: The JN-DSRCT-1 cell line, in vitro, is sensitive to trabectedin: after drug exposure, EWS-WT1 chimera expression decreases as well as binding on its target promoters. Probably the heterogeneity of chimera transcripts is an obstacle to precisely defining the molecular mode of action of drugs, calling for further cellular models of DSRCT, possibly growing in vivo too, to mimic the biological complexity of this disease.",
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AU - Craparotta, I.

AU - Colella, G.

AU - Ronchetti, E.

AU - Beltrame, L.

AU - Vicario, S.

AU - Marchini, S.

AU - Panini, N.

AU - Dagrada, G.

AU - Bozzi, F.

AU - Pilotti, S.

AU - Galmarini, C. M.

AU - D'Incalci, M.

AU - Gatta, R.

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N2 - Background: Desmoplastic small round cell tumor (DSRCT) is a rare and highly aggressive disease, that can be described as a member of the family of small round blue cell tumors. The molecular diagnostic marker is the t(11;22)(p13;q12) translocation, which creates an aberrant transcription factor, EWS-WT1, that underlies the oncogenesis of DSRCT. Current treatments are not very effective so new active drugs are needed. Trabectedin, now used as a single agent for the treatment of soft tissue sarcoma, was reported to be active in some pre-treated DSRCT patients. Using JN-DSRCT-1, a cell line derived from DSRCT expressing the EWS-WT1 fusion protein, we investigated the ability of trabectedin to modify the function of the chimeric protein, as in other sarcomas expressing fusion proteins. After detailed characterization of the EWS-WT1 transcripts structure, we investigated the mode of action of trabectedin, looking at the expression and function of the oncogenic chimera. Methods: We characterized JN-DSRCT-1 cells using cellular approaches (FISH, Clonogenicity assay) and molecular approaches (Sanger sequencing, ChIP, GEP). Results: JN-DSRCT-1 cells were sensitive to trabectedin at nanomolar concentrations. The cell line expresses different variants of EWS-WT1, some already identified in patients. EWS-WT1 mRNA expression was affected by trabectedin and chimeric protein binding on its target gene promoters was reduced. Expression profiling indicated that trabectedin affects the expression of genes involved in cell proliferation and apoptosis. Conclusions: The JN-DSRCT-1 cell line, in vitro, is sensitive to trabectedin: after drug exposure, EWS-WT1 chimera expression decreases as well as binding on its target promoters. Probably the heterogeneity of chimera transcripts is an obstacle to precisely defining the molecular mode of action of drugs, calling for further cellular models of DSRCT, possibly growing in vivo too, to mimic the biological complexity of this disease.

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