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Isolation of extracellular vesicles improves the detection of mutant DNA from plasma of metastatic melanoma patients
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Isolation of extracellular vesicles improves the detection of mutant DNA from plasma of metastatic melanoma patients
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Journal Article
Isolation of extracellular vesicles improves the detection of mutant DNA from plasma of metastatic melanoma patients
2020
Overview
Detection of
BRAF
V600E
within cell free tumor DNA (ctDNA) is emerging as a promising means to improve patients’ stratification or enable BRAF inhibitor (BRAFi) therapeutic monitoring in a minimally invasive manner. Here, we investigated whether extracellular vesicle-(EV)-associated-DNA (EV-DNA) has value as an alternative source of circulating
BRAF
V600E
. To do so, we identified a clinical practice-compatible protocol for the isolation of EV-DNA and assessed
BRAF
gene status on plasma samples from metastatic melanoma patients at the beginning and during BRAFi therapy. This protocol uses a peptide with high affinity for EVs and it has been found to recover more mutant DNA from plasma than standard ultracentrifugation. Molecular analyses revealed that mutant DNA is largely unprotected from nuclease digestion, interacting with the outer side of the EV membrane or directly with the peptide. When used on clinical samples, we found that the protocol improves the detection of
BRAF
V600E
gene copies in comparison to the reference protocol for ctDNA isolation. Taken together, these findings indicate that EVs are a promising source of mutant DNA and should be considered for the development of next-generation liquid biopsy approaches.
Publisher
Nature Publishing Group UK
Subject
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