학술논문
Proof-of-Concept for Liquid Biopsy Disease Monitoring of MYC -Amplified Group 3 Medulloblastoma by Droplet Digital PCR.
Document Type
Article
Author
Stepien, Natalia; Senfter, Daniel; Furtner, Julia; Haberler, Christine; Dorfer, Christian; Czech, Thomas; Lötsch-Gojo, Daniela; Mayr, Lisa; Hedrich, Cora; Baumgartner, Alicia; Aliotti-Lippolis, Maria; Schned, Hannah; Holler, Johannes; Bruckner, Katharina; Slavc, Irene; Azizi, Amedeo A.; Peyrl, Andreas; Müllauer, Leonhard; Madlener, Sibylle; Gojo, Johannes
Source
Subject
*PUBLIC health surveillance
*DISEASE progression
*GLIOMAS
*FLUORESCENCE in situ hybridization
*RESEARCH funding
*POLYMERASE chain reaction
*SENSITIVITY & specificity (Statistics)
*DATA analysis software
*LONGITUDINAL method
BODY fluid examination
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Language
ISSN
2072-6694
Abstract
Simple Summary: MYC is a well-described oncogene across multiple cancer types. In medulloblastoma (MB), significance is subtype-dependent and associated with a particularly dismal outcome in MYC-amplified group 3 MBs. In our study, we established a highly sensitive and specific method for the detection of MYC amplification in cerebrospinal fluid (CSF) enabling its use as a liquid biopsy biomarker. We show preliminary results of its potential as a marker for early diagnosis, disease staging and monitoring. The fast and cost-effective method could substantially improve means of diagnosis and therapy monitoring in high-risk MBs. Background: Liquid biopsy diagnostic methods are an emerging complementary tool to imaging and pathology techniques across various cancer types. However, there is still no established method for the detection of molecular alterations and disease monitoring in MB, the most common malignant CNS tumor in the pediatric population. In the presented study, we investigated droplet digital polymerase chain reaction (ddPCR) as a highly sensitive method for the detection of MYC amplification in bodily fluids of group 3 MB patients. Methods: We identified a cohort of five MYC-amplified MBs by methylation array and FISH. Predesigned and wet-lab validated probes for ddPCR were used to establish the detection method and were validated in two MYC-amplified MB cell lines as well as tumor tissue of the MYC-amplified cohort. Finally, a total of 49 longitudinal CSF samples were analyzed at multiple timepoints during the course of the disease. Results: Detection of MYC amplification by ddPCR in CSF showed a sensitivity and specificity of 90% and 100%, respectively. We observed a steep increase in amplification rate (AR) at disease progression in 3/5 cases. ddPCR was proven to be more sensitive than cytology for the detection of residual disease. In contrast to CSF, MYC amplification was not detectable by ddPCR in blood samples. Conclusions: ddPCR proves to be a sensitive and specific method for the detection of MYC amplification in the CSF of MB patients. These results warrant implementation of liquid biopsy in future prospective clinical trials to validate the potential for improved diagnosis, disease staging and monitoring. [ABSTRACT FROM AUTHOR]