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Precision diagnostics of Ewing’s sarcoma by liquid biopsy: circulating EWS-FLI1 fusion transcripts
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Journal Article
Precision diagnostics of Ewing’s sarcoma by liquid biopsy: circulating EWS-FLI1 fusion transcripts
2018
Overview
Background:
Limited information is available on the applicative value of liquid biopsy (LB) in rare
tumors, including Ewing’s sarcoma (ES). The accepted precision diagnostics standards
would greatly benefit from a non-invasive LB test monitoring pathognomonic gene
rearrangements in the bloodstream.
Methods:
Tissue and blood samples were collected from six and four ES patients, respectively.
Plasma was cleared by two successive rounds of centrifugation and stored frozen until
RNA extraction by the QIAmp CNA kit. RNA was retro-transcribed and subjected to
real-time quantitative polymerase chain reaction (RT-qPCR) and digital polymerase chain
reaction (dPCR). Reactions were set up using two custom primer sets identifying types 1
and 2 EWS-FLI1 fusion transcripts.
Results:
The two prevalent types of EWS-FLI1 rearrangements could be identified
using only two sets of polymerase chain reaction primers, regardless of patient-specific
EWS-FLI1 DNA breakpoints. RT-qPCR and dPCR discriminated the two
variants in five tumor tissue RNAs and in four circulating tumor RNAs (ctRNAs). Of note,
EWS-FLI1 molecular diagnosis was possible using blood samples even
when tumor tissue was not available. ctRNA levels correlated (p <
0.05) with volume-based positron emission tomography (PET) parameters (metabolic tumor
volume and total lesion glycolysis), and allowed the fine tracking of disease course
after surgery, during adjuvant as well as neoadjuvant chemotherapy, and at follow up in
one patient.
Conclusions:
To our knowledge, this is one of the few single-marker LB assays in solid tumors
specifically designed to detect rearranged RNAs in blood, and the first study describing
EWS circulating tumor RNAs in ES patients. Altogether, our results
support the idea that LB may have a considerable impact on ES patient monitoring and
management.
Publisher
SAGE Publications,SAGE Journals
Subject
