BRAF V600E Detection in Liquid Biopsies from Pediatric Central Nervous System Tumors

Abstract

Pediatric Central Nervous System (CNS) tumors are the most fatal cancer diseases in childhood. Due to their localization and infiltrative nature, some tumor resections or biopsies are not feasible. In those cases, the use of minimally invasive methods as diagnostic, molecular marker detection, prognostic or monitoring therapies are emerging. The analysis of liquid biopsies which contain genetic information from the tumor has been much more widely explored in adults than in children. We compare the detection of BRAF V600E targetable mutation by digital-PCR from cell-free-DNA and EV-derived DNA (ctDNA) in serum, plasma and cerebrospinal fluid (CSF) isolated from a cohort of 29 CNS pediatric patients. Here we demonstrate that ctDNA isolated from serum and plasma could be successfully analyzed to obtain tumor genetic information which could be used to guide critical treatment decisions.

Multiplexed Droplet Digital PCR Assays for the Simultaneous Screening of Major Genetic Alterations in Tumors of the Central Nervous System

“…Therefore, multiplexed dPCR assays would be specifically appropriate for histomolecular classification of gliomas and glioneuronal tumors as most laboratories are likely to have the capacities to use dPCR technology when compared to NGS. Furthermore, dPCR can also be used in many applications requiring high sensitivity, particularly for the detection of circulating tumor DNA in blood plasma or cerebrospinal fluid (24).…”

Liquid Biopsy is Instrumental for 3PM Dimensional Solutions in Cancer Management

“…Despite great potential and many advantages over tissue biopsy, the utilization of liquid biopsy biomarkers is still associated with various challenges. High accuracy of biomarker detection within the scope of liquid biopsy is one of the most important challenges in clinical practice [ 39 ]. The lack of implementation of liquid biopsy into standard clinical practice, which is related to the unavailability of standardized techniques and guidelines, especially in terms of collection of samples and pre-analytical conditions, [ 26 , 40 ] can be illustrated by an example of cell-free DNA evaluation from blood and urine samples.…”

“…Despite the fact that a higher concentration of cell-free DNA is generally associated with serum, it is not a preferable liquid biopsy sample because of the contamination during the storage procedure [ 42 , 102 ] due to potentially high content of DNA released from lysis of circulating blood cells [ 103 ]. However, the comparison of the detection of the BRAF V600E-targetable mutation by dPCR from ctDNA and EV-derived DNA in plasma, serum, and CSF revealed serum to be more promising than plasma in liquid biopsy from pediatric central nervous tumors [ 39 ]. Above all, liquid biopsy profiling using ctDNA establishes an appropriate therapeutic approach, assessment of real-time conditions on cancer patients, and is associated with a potential of early detection of cancer [ 103 ].…”

Utility of a bridged nucleic acid clamp for liquid biopsy: Detecting BRAF V600E in the cerebrospinal fluid of a patient with brain tumor

“…Liquid biopsy is a promising approach for detecting tumor‐specific abnormalities without the risk of surgical complications 1‐5 . Herein, we demonstrate the utility of bridged nucleic acid (BNA) for liquid biopsy and report a case of a brain tumor wherein BRAF V600E was successfully detected in the cerebrospinal fluid (CSF).…”