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Updated: Apr 11, 2026

Evaluating In Vitro DNA Damage Using Comet Assay
Published on: October 11, 2017
The PARP inhibitor olaparib enhances the sensitivity of Ewing sarcoma to trabectedin
José Luis Ordóñez1,2, Ana Teresa Amaral1,2, Angel M Carcaboso3
1Laboratory of Molecular Pathology, Instituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocio/CSIC/Universidad de Sevilla, Seville, Spain.
Abstract:
Recent preclinical evidence has suggested that Ewing Sarcoma (ES) bearing EWSR1-ETS fusions could be particularly sensitive to PARP inhibitors (PARPinh) in combination with DNA damage repair (DDR) agents. Trabectedin is an antitumoral agent that modulates EWSR1-FLI1 transcriptional functions, causing DNA damage. Interestingly, PARP1 is also a transcriptional regulator of EWSR1-FLI1, and PARPinh disrupts the DDR machinery. Thus, given the impact and apparent specificity of both agents with regard to the DNA damage/DDR system and EWSR1-FLI1 activity in ES, we decided to explore the activity of combining PARPinh and Trabectedin in in vitro and in vivo experiments. The combination of Olaparib and Trabectedin was found to be highly synergistic, inhibiting cell proliferation, inducing apoptosis, and the accumulation of G2/M. The drug combination also enhanced γH2AX intranuclear accumulation as a result of DNA damage induction, DNA fragmentation and global DDR deregulation, while EWSR1-FLI1 target expression remained unaffected. The effect of the drug combination was corroborated in a mouse xenograft model of ES and, more importantly, in two ES patient-derived xenograft (PDX) models in which the tumors showed complete regression. In conclusion, the combination of the two agents leads to a biologically significant deregulation of the DDR machinery that elicits relevant antitumor activity in preclinical models and might represent a promising therapeutic tool that should be further explored for translation to the clinical setting.
Insights
Combining PARP inhibitors with Trabectedin shows significant synergy against Ewing Sarcoma (ES). This DNA damage repair (DDR) targeting strategy induces tumor regression in preclinical models, offering a promising new therapeutic avenue for ES patients.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Ewing Sarcoma (ES) with EWSR1-ETS fusions may respond to PARP inhibitors (PARPinh) and DNA damage repair (DDR) agents.
- Trabectedin damages DNA and affects EWSR1-FLI1 transcription; PARP1 regulates EWSR1-FLI1 and PARPinh disrupts DDR.
Purpose of the Study:
- To investigate the synergistic activity of combining PARP inhibitors and Trabectedin in ES.
- To evaluate the in vitro and in vivo efficacy of this drug combination.
Main Methods:
- In vitro studies assessing cell proliferation, apoptosis, and cell cycle.
- In vivo experiments using ES mouse xenograft and patient-derived xenograft (PDX) models.
- Analysis of DNA damage markers (γH2AX) and DDR pathway deregulation.
Main Results:
- Olaparib and Trabectedin combination demonstrated high synergy, inhibiting proliferation and inducing apoptosis.
- The combination enhanced DNA damage (γH2AX accumulation) and deregulated global DDR.
- Complete tumor regression was observed in ES PDX models.
Conclusions:
- The combination of PARP inhibitors and Trabectedin causes significant DDR deregulation and potent antitumor activity in preclinical ES models.
- This combination represents a promising therapeutic strategy for Ewing Sarcoma, warranting clinical investigation.
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