Targeted Analysis of Cell-free Circulating Tumor DNA is Suitable for Early Relapse and Actionable Target Detection in Patients with Neuroblastoma
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Updated: Oct 1, 2025
Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids
Published on: June 8, 2019
Marco Lodrini1,2,3, Josefine Graef4, Theresa M Thole-Kliesch1
1Department of Pediatric Hematology and Oncology, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Purpose:
Treating refractory or relapsed neuroblastoma remains challenging. Monitoring body fluids for tumor-derived molecular information indicating minimal residual disease supports more frequent diagnostic surveillance and may have the power to detect resistant subclones before they give rise to relapses. If actionable targets are identified from liquid biopsies, targeted treatment options can be considered earlier.
Experimental Design:
Droplet digital PCR assays assessing MYCN and ALK copy numbers and allelic frequencies of ALK p.F1174L and ALK p.R1275Q mutations were applied to longitudinally collected liquid biopsies and matched tumor tissue samples from 31 patients with high-risk neuroblastoma. Total cell-free DNA (cfDNA) levels and marker detection were compared with data from routine clinical diagnostics.
Results:
Total cfDNA concentrations in blood plasma from patients with high-risk neuroblastoma were higher than in healthy controls and consistently correlated with neuron-specific enolase levels and lactate dehydrogenase activity but not with 123I-meta-iodobenzylguanidine scores at relapse diagnosis. Targeted cfDNA diagnostics proved superior for early relapse detection to all current diagnostics in 2 patients. Marker analysis in cfDNA indicated intratumor heterogeneity for cell clones harboring MYCN amplifications and druggable ALK alterations that were not detectable in matched tumor tissue samples in 17 patients from our cohort. Proof of concept is provided for molecular target detection in cerebrospinal fluid from patients with isolated central nervous system relapses.
Conclusions:
Tumor-specific alterations can be identified and monitored during disease course in liquid biopsies from pediatric patients with high-risk neuroblastoma. This approach to cfDNA surveillance warrants further prospective validation and exploitation for diagnostic purposes and to guide therapeutic decisions.
Insights
Monitoring liquid biopsies for neuroblastoma can detect resistant subclones early. This approach using cell-free DNA (cfDNA) surveillance aids in timely diagnosis and targeted treatment decisions for high-risk patients.
Area of Science:
- Pediatric Oncology
- Molecular Diagnostics
- Liquid Biopsy Analysis
Background:
- Refractory or relapsed neuroblastoma treatment is challenging.
- Early detection of minimal residual disease and resistant subclones is crucial for improved patient outcomes.
- Liquid biopsies offer a non-invasive method to monitor tumor-derived molecular information.
Purpose of the Study:
- To assess the utility of droplet digital PCR (ddPCR) assays on liquid biopsies for monitoring high-risk neuroblastoma.
- To compare cell-free DNA (cfDNA) marker detection with routine clinical diagnostics for early relapse detection.
- To investigate intratumor heterogeneity and identify actionable targets in neuroblastoma.
Main Methods:
- Longitudinal liquid biopsies and matched tumor tissue from 31 high-risk neuroblastoma patients were analyzed.
- Droplet digital PCR (ddPCR) was used to assess MYCN and ALK copy numbers and mutations in cfDNA.
- Total cfDNA levels and marker detection were correlated with clinical data and routine diagnostics.
Main Results:
- Elevated cfDNA concentrations were observed in neuroblastoma patients compared to healthy controls.
- cfDNA diagnostics demonstrated superiority in early relapse detection in two patients.
- Intratumor heterogeneity, including MYCN amplifications and druggable ALK alterations, was identified in cfDNA from 17 patients, often undetectable in tumor tissue.
- Molecular target detection in cerebrospinal fluid for central nervous system relapses was successfully demonstrated.
Conclusions:
- Tumor-specific alterations can be identified and monitored non-invasively through liquid biopsies in pediatric neuroblastoma.
- cfDNA surveillance is a promising approach for diagnostic purposes and guiding therapeutic decisions in high-risk neuroblastoma.
- Further prospective validation of cfDNA surveillance is warranted for clinical application.
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