title: Plasma vesicle miRNAs for therapy response monitoring in Hodgkin lymphoma patients.
abstract: BACKGROUND.Cell-free circulating nucleic acids, including 22-nt microRNAs (miRNAs), represent noninvasive biomarkers for treatment response monitoring of cancer patients. While the majority of plasma miRNA is bound to proteins, a smaller, less well-characterized pool is associated with extracellular vesicles (EVs). Here, we addressed whether EV-associated miRNAs reflect metabolic disease in classical Hodgkin lymphoma (cHL) patients.METHODS.With standardized size-exclusion chromatography (SEC), we isolated EV-associated extracellular RNA (exRNA) fractions and protein-bound miRNA from plasma of cHL patients and healthy subjects. We performed a comprehensive small RNA sequencing analysis and validation by TaqMan qRT-PCR for candidate discovery. Fluorodeoxyglucose-PET (FDG-PET) status before treatment, directly after treatment, and during long-term follow-up was compared directly with EV miRNA levels.RESULTS.The plasma EV miRNA repertoire was more extensive compared with protein-bound miRNA that was heavily dominated by a few abundant miRNA species and was less informative of disease status. Purified EV fractions of untreated cHL patients and tumor EVs had enriched levels of miR24-3p, miR127-3p, miR21-5p, miR155-5p, and let7a-5p compared with EV fractions from healthy subjects and disease controls. Serial monitoring of EV miRNA levels in patients before treatment, directly after treatment, and during long-term follow-up revealed robust, stable decreases in miRNA levels matching a complete metabolic response, as observed with FDG-PET. Importantly, EV miRNA levels rose again in relapse patients.CONCLUSION.We conclude that cHL-related miRNA levels in circulating EVs reflect the presence of vital tumor tissue and are suitable for therapy response and relapse monitoring in individual cHL patients.FUNDING.Cancer Center Amsterdam Foundation (CCA-2013), Dutch Cancer Society (KWF-5510), Technology Foundation STW (STW Perspectief CANCER-ID).
authors.affiliation: Department of Pathology, Exosomes Research Group, and.
authors.forename: Monique Aj
authors.initials: MA
authors.lastname: van Eijndhoven
authors.affiliation: Department of Hematology, VU University Medical Center, Amsterdam, Netherlands.
authors.forename: Josée M
authors.initials: JM
authors.lastname: Zijlstra
authors.affiliation: Department of Pathology, Exosomes Research Group, and.
authors.forename: Nils J
authors.initials: NJ
authors.lastname: Groenewegen
authors.affiliation: Department of Pathology, Exosomes Research Group, and.
authors.forename: Esther Ee
authors.initials: EE
authors.lastname: Drees
authors.affiliation: Department of Pathology, Exosomes Research Group, and.
authors.forename: Stuart
authors.initials: S
authors.lastname: van Niele
authors.affiliation: Department of Pathology, Exosomes Research Group, and.
authors.forename: S Rubina
authors.initials: SR
authors.lastname: Baglio
authors.affiliation: Department of Pathology, Exosomes Research Group, and.
authors.forename: Danijela
authors.initials: D
authors.lastname: Koppers-Lalic
authors.affiliation: iZON Science, Oxford, United Kingdom.
authors.forename: Hans
authors.initials: H
authors.lastname: van der Voorn
authors.affiliation: Department of Biochemistry and Cell Biology, Utrecht University, Utrecht, Netherlands.
authors.forename: Sten Fwm
authors.initials: SF
authors.lastname: Libregts
authors.affiliation: Department of Biochemistry and Cell Biology, Utrecht University, Utrecht, Netherlands.
authors.forename: Marca Hm
authors.initials: MH
authors.lastname: Wauben
authors.affiliation: Department of Epidemiology and Biostatistics, VU University Medical Center, Amsterdam, Netherlands.
authors.forename: Renee X
authors.initials: RX
authors.lastname: de Menezes
authors.affiliation: Department of Functional Genomics, Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam, VU University, Amsterdam, Netherlands.
authors.forename: Jan Rt
authors.initials: JR
authors.lastname: van Weering
authors.affiliation: Department of Clinical Chemistry, Academic Medical Center, Amsterdam, Netherlands.
authors.forename: Rienk
authors.initials: R
authors.lastname: Nieuwland
authors.affiliation: Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Groningen, Netherlands.
authors.forename: Lydia
authors.initials: L
authors.lastname: Visser
authors.affiliation: Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Groningen, Netherlands.
authors.forename: Anke
authors.initials: A
authors.lastname: van den Berg
authors.affiliation: Department of Pathology, Exosomes Research Group, and.
authors.forename: Daphne
authors.initials: D
authors.lastname: de Jong
authors.affiliation: Department of Pathology, Exosomes Research Group, VU University Medical Center, Amsterdam, Netherlands; ExBiome BV, Amsterdam, Netherlands.
authors.forename: D Michiel
authors.initials: DM
authors.lastname: Pegtel
mesh_terms: D014408:Biomarkers, Tumor / Q000097:blood; D016022:Case-Control Studies; D000067128:Extracellular Vesicles; D006689:Hodgkin Disease / Q000097:blood* / Q000628:therapy; D006801:Humans; D035683:MicroRNAs / Q000097:blood*; D009364:Neoplasm Recurrence, Local
chemical_list: D014408:Biomarkers, Tumor; D035683:MicroRNAs