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Updated: Aug 18, 2025
Cytotoxic Efficacy of Photodynamic Therapy in Osteosarcoma Cells In Vitro
Published on: March 18, 2014
Establishment of Rapid and Accurate Screening System for Molecular Target Therapy of Osteosarcoma
Keita Sasa1,2, Tsuyoshi Saito2,3, Taisei Kurihara1
1Department of Medicine for Orthopaedics and Motor Organ, Juntendo University School of Medicine, Tokyo, Japan.
Abstract:
Introduction Comprehensive analyses using clinical sequences subcategorized osteosarcoma (OS) into several groups according to the activated signaling pathways. Mutually exclusive co-occurrences of gene amplification (PDGFRA/KIT/KDR, VEGFA/CCND3, and MDM2/CDK4) have been identified in approximately 40% of OS, representing candidate subsets for clinical evaluation of additional therapeutic options. Thus, it would be desirable to evaluate the specific gene amplification before starting therapy in patients with OS. Materials and Methods This is a retrospective study. We examined 13 cases of clinical OS samples using NanoString-based copy number variation (CNV) analysis. Decalcification and chemotherapeutic effects on this analysis were also assessed. Results First, the accuracy of this system was validated by showing that amplification/deletion data obtained from this system using various types of cancer cell lines almost perfectly matched to that from the Cancer Cell Line Encyclopedia (CCLE). We identified potentially actionable alterations in CDK4/MDM2 amplification in 10% of samples and potential additional therapeutic targets (PDGFRA/KIT/KDR and VEGFA/CCND3) in 20% of samples, which is consistent with the reported frequencies. Furthermore, this assay could identify these potential therapeutic targets regardless of the sample status (frozen vs formalin-fixed paraffin-embedded [FFPE] tissues). Conclusion We established a NanoString-based rapid and cost-effective method with a short turnaround time (TAT) to examine gene amplification status in OS. This CNV analysis using FFPE samples is recommended where the histological evaluation of viable tumor cells is possible, especially for tumors after chemotherapy with higher chemotherapeutic effects.
Insights
A new NanoString-based method accurately detects gene amplifications in osteosarcoma (OS) using formalin-fixed paraffin-embedded (FFPE) samples. This rapid, cost-effective test identifies potential therapeutic targets, aiding personalized treatment strategies for OS patients.
Area of Science:
- Oncology
- Genetics
- Molecular Biology
Background:
- Osteosarcoma (OS) is subcategorized based on activated signaling pathways, with specific gene amplifications identified in about 40% of cases.
- Mutually exclusive co-occurrences of gene amplifications, including PDGFRA/KIT/KDR, VEGFA/CCND3, and MDM2/CDK4, represent potential therapeutic targets in OS.
- Accurate and timely assessment of gene amplification status is crucial for guiding therapeutic decisions in OS patients.
Purpose of the Study:
- To establish a rapid, cost-effective NanoString-based method for assessing gene amplification status in osteosarcoma.
- To evaluate the feasibility of using formalin-fixed paraffin-embedded (FFPE) samples for copy number variation (CNV) analysis in OS.
- To identify potentially actionable gene amplifications for targeted therapy in OS.
Main Methods:
- Retrospective study involving 13 clinical OS samples.
- NanoString-based copy number variation (CNV) analysis to detect gene amplifications and deletions.
- Assessment of decalcification and chemotherapeutic effects on the NanoString assay accuracy.
Main Results:
- The NanoString system demonstrated high accuracy, closely matching data from the Cancer Cell Line Encyclopedia (CCLE).
- Potentially actionable alterations (CDK4/MDM2 amplification) were identified in 10% of samples.
- Additional therapeutic targets (PDGFRA/KIT/KDR and VEGFA/CCND3) were found in 20% of samples, consistent with previous findings.
- The assay successfully identified targets in both frozen and FFPE samples, irrespective of pre-analytical treatments.
Conclusions:
- A rapid, cost-effective NanoString-based CNV analysis method was successfully established for OS gene amplification profiling.
- The method is suitable for use with FFPE samples, particularly those from post-chemotherapy tumors where viable cells can be assessed.
- This assay provides a valuable tool for identifying therapeutic targets and guiding personalized treatment strategies in osteosarcoma.
