Extracellular Vesicle-Based Method for Detecting MYCN Amplification Status of Pediatric Neuroblastoma
3
Issued Date
2022-06-01
Resource Type
eISSN
20726694
Scopus ID
2-s2.0-85131012174
Journal Title
Cancers
Volume
14
Issue
11
Rights Holder(s)
SCOPUS
Bibliographic Citation
Cancers Vol.14 No.11 (2022)
Suggested Citation
Panachan J., Rojsirikulchai N., Pongsakul N., Khowawisetsut L., Pongphitcha P., Siriboonpiputtana T., Chareonsirisuthigul T., Phornsarayuth P., Klinkulab N., Jinawath N., Chiangjong W., Anurathapan U., Pattanapanyasat K., Hongeng S., Chutipongtanate S. Extracellular Vesicle-Based Method for Detecting MYCN Amplification Status of Pediatric Neuroblastoma. Cancers Vol.14 No.11 (2022). doi:10.3390/cancers14112627 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/83721
Title
Extracellular Vesicle-Based Method for Detecting MYCN Amplification Status of Pediatric Neuroblastoma
Other Contributor(s)
Abstract
MYCN amplification is the strongest predictor of high-risk neuroblastoma (NB). The standard procedure to detect MYCN status requires invasive procedures. Extracellular vesicles (EVs) contain molecular signatures of originated cells, present in biofluids, and serve as an invaluable source for cancer liquid biopsies. This study aimed to establish an EV-based method to detect the MYCN status of NB. Two EV subtypes, i.e., microvesicles (MVs) and exosomes, were sequentially isolated from the culture supernatant by step-wise centrifugation, ultrafiltration, and size-exclusion chromatography. Quantitative RT-PCR was performed to detect MYCN mRNA. As a result, MYCN mRNA was detectable in the MVs, but not exosomes, of MYCN-amplified NB cells. MYCN mRNA-containing MVs (MYCN-MV) were successfully detected in three distinct MYCN-amplified NB cell lines but absent in three MYCN non-amplification cells. The simulated samples were prepared by pulsing MVs into human serum. MYCN–MV detection in the simulated samples showed a less interfering effect from the human blood matrix. Validation using clinical specimens (2 mL bone marrow plasma) obtained from patients at various disease stages showed a promising result. Five out of six specimens of MYCN-amplified patients showed positive results, while there were no false positives in four plasma samples of the MYCN non-amplification group. This study communicated a novel EV-based method for detecting the MYCN status of pediatric NB based on MYCN mRNA contents in MVs. Future studies should be pursued in a prospective cohort to determine its true diagnostic performance.