Plasma vesicle miRNAs for therapy response monitoring in Hodgkin lymphoma patients
Monique A.J. van Eijndhoven, … , Daphne de Jong, D. Michiel Pegtel
Monique A.J. van Eijndhoven, … , Daphne de Jong, D. Michiel Pegtel
Published November 17, 2016
Citation Information: JCI Insight. 2016;1(19):e89631. https://doi.org/10.1172/jci.insight.89631.
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Clinical Research and Public Health Hematology Oncology

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

Monique A.J. van Eijndhoven, Josée M. Zijlstra, Nils J. Groenewegen, Esther E.E. Drees, Stuart van Niele, S. Rubina Baglio, Danijela Koppers-Lalic, Hans van der Voorn, Sten F.W.M. Libregts, Marca H.M. Wauben, Renee X. de Menezes, Jan R.T. van Weering, Rienk Nieuwland, Lydia Visser, Anke van den Berg, Daphne de Jong, D. Michiel Pegtel

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Figure 3

RNAseq reveals lymphoma-secreted miRNAs in circulating extracellular vesicles.

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RNAseq reveals lymphoma-secreted miRNAs in circulating extracellular ves...
(A) RNAseq analysis of miR24-3p, miR127-3p, miR155-5p, miR21-5p, and let7a-5p in extracellular vesicles (EVs) isolated from B cell lymphoma cell lines. Data are shown as reads per million miRNA reads (RPM). (B) Number of different miRNAs identified in plasma EV and protein/HDL fractions of healthy donors and cHL patients (n = 3 each). *P < 0.05 (unpaired 2-tailed t test). (C) Distribution of small RNA subclasses in plasma EVs of a healthy individual and a cHL patient. Data shown are of 1 representative donor (n = 3) and depicted as percentage read counts. (D) RNAseq analysis of let7a-5p, miR21-5p, miR127-3p, and miR486-5p in healthy and cHL patient plasma EVs and protein/HDL fractions. Dots represent individual samples; error bars represent mean ± SEM. Reads are normalized using the trimmed mean of M values (TMM) method. (E) As in D, but data are shown as summed read counts.