TY - JOUR
T1 - Cell-Free RNA from Plasma in Patients with Neuroblastoma
T2 - Exploring the Technical and Clinical Potential
AU - Lak, Nathalie S M
AU - Seijger, Anne
AU - van Zogchel, Lieke M J
AU - Gelineau, Nina U
AU - Javadi, Ahmad
AU - Zappeij-Kannegieter, Lily
AU - Bongiovanni, Laura
AU - Andriessen, Anneloes
AU - Stutterheim, Janine
AU - van der Schoot, C Ellen
AU - de Bruin, Alain
AU - Tytgat, Godelieve A M
N1 - Funding Information:
N.S.M.L.: L.Z.-K. and J.S. were supported by grant 312, from Children Cancer-Free (KiKa).
Publisher Copyright:
© 2023 by the authors.
PY - 2023/4/1
Y1 - 2023/4/1
N2 - Neuroblastoma affects mostly young children, bearing a high morbidity and mortality. Liquid biopsies, e.g., molecular analysis of circulating tumor-derived nucleic acids in blood, offer a minimally invasive diagnostic modality. Cell-free RNA (cfRNA) is released by all cells, especially cancer. It circulates in blood packed in extracellular vesicles (EV) or attached to proteins. We studied the feasibility of analyzing cfRNA and EV, isolated by size exclusion chromatography (SEC), from platelet-poor plasma from healthy controls (
n = 40) and neuroblastoma patients with localized (
n = 10) and metastatic disease (
n = 30). The mRNA content was determined using several multiplex droplet digital PCR (ddPCR) assays for a neuroblastoma-specific gene panel (
PHOX2B,
TH,
CHRNA3) and a cell cycle regulation panel (
E2F1,
CDC6,
ATAD2,
H2AFZ,
MCM2,
DHFR). We applied corrections for the presence of platelets. We demonstrated that neuroblastoma-specific markers were present in plasma from 14/30 patients with metastatic disease and not in healthy controls and patients with localized disease. Most cell cycle markers had a higher expression in patients. The mRNA markers were mostly present in the EV-enriched SEC fractions. In conclusion, cfRNA can be isolated from plasma and EV and analyzed using multiplex ddPCR. cfRNA is an interesting novel liquid biopsy-based target to explore further.
AB - Neuroblastoma affects mostly young children, bearing a high morbidity and mortality. Liquid biopsies, e.g., molecular analysis of circulating tumor-derived nucleic acids in blood, offer a minimally invasive diagnostic modality. Cell-free RNA (cfRNA) is released by all cells, especially cancer. It circulates in blood packed in extracellular vesicles (EV) or attached to proteins. We studied the feasibility of analyzing cfRNA and EV, isolated by size exclusion chromatography (SEC), from platelet-poor plasma from healthy controls (
n = 40) and neuroblastoma patients with localized (
n = 10) and metastatic disease (
n = 30). The mRNA content was determined using several multiplex droplet digital PCR (ddPCR) assays for a neuroblastoma-specific gene panel (
PHOX2B,
TH,
CHRNA3) and a cell cycle regulation panel (
E2F1,
CDC6,
ATAD2,
H2AFZ,
MCM2,
DHFR). We applied corrections for the presence of platelets. We demonstrated that neuroblastoma-specific markers were present in plasma from 14/30 patients with metastatic disease and not in healthy controls and patients with localized disease. Most cell cycle markers had a higher expression in patients. The mRNA markers were mostly present in the EV-enriched SEC fractions. In conclusion, cfRNA can be isolated from plasma and EV and analyzed using multiplex ddPCR. cfRNA is an interesting novel liquid biopsy-based target to explore further.
KW - cell-free RNA
KW - extracellular vesicles
KW - liquid biopsies
KW - neuroblastoma
KW - pediatric
KW - solid tumors
UR - http://www.scopus.com/inward/record.url?scp=85152948430&partnerID=8YFLogxK
U2 - 10.3390/cancers15072108
DO - 10.3390/cancers15072108
M3 - Article
C2 - 37046768
SN - 2072-6694
VL - 15
SP - 1
EP - 16
JO - Cancers
JF - Cancers
IS - 7
M1 - 2108
ER -