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Extracellular vesicle-associated miRNAs in ovarian cancer

Design of an integrated NGS-based workflow for the identification of blood-based biomarkers for platinum-resistance
 
: Kuhlmann, Jan Dominik; Chebouti, Issam; Kimmig, Rainer; Buderath, Paul; Reuter, Michael; Puppel, Sven Holger; Wimberger, Pauline; Kasimir-Bauer, Sabine

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Clinical chemistry. Journal of the American Association for Clinical Chemistry 57 (2019), No.7, pp.1053-1062
ISSN: 0009-9147
ISSN: 1530-8561
English
Journal Article
Fraunhofer IZI ()

Abstract
Extracellular vesicle (EV)-associated microRNAs (miRNAs) have been suggested as promising biomarkers for blood-based cancer diagnosis. However, one of the major limitations for the use of EVs with diagnostic purpose is the lack of standardized EV-profiling techniques. In this regard, the objective of our study was to design an integrated next-generation sequencing (NGS)-based workflow for analyzing the signature of EV-associated miRNA in the plasma of platinum-resistant ovarian cancer patients. For EV-extraction, different enrichment methods were compared (ExoQuick vs. exoRNeasy). NGS was performed with the Illumina platform. We established an integrated NGS-based workflow, including EV-enrichment with the ExoQuick system, which resulted in an optimal RNA-yield and consistent small RNA libraries. We applied this workflow in a pilot cohort of clinically documented platinum-sensitive (n=15) vs. platinum-resistant (n=15) ovarian cancer patients, resulting in a panel of mature EV-associated miRNAs (including ovarian cancer associated miR-181a, miR-1908, miR-21, miR-486 and miR-223), which were differentially abundant in the plasma of platinum-resistant patients. This is the first study, analyzing the profile of EV-associated miRNAs in platinum-resistant ovarian cancer patients. We provide rationale to further validate these miRNA candidates in an independent set of patients, in order to characterize their biomarker potential as predictors for platinum-resistance.

: http://publica.fraunhofer.de/documents/N-534720.html