Plasma vesicle miRNAs for therapy response monitoring in Hodgkin lymphoma patients

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

doi:10.1172/jci.insight.89631

Cited by 143 publications

(184 citation statements)

References 50 publications

(92 reference statements)

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“…Although we observed lower levels of miR-21-5p in mixed cellularity compared to nodular sclerosis cHL cell lines, this was not supported by a previous study showing similar miR-21-5p levels in nodular sclerosis and mixed cellularity cell lines and cHL total tissues [18]. More recently, plasma miR-21-5p levels were identified as a potential circulating biomarker in cHL patients [48,49]. A moderate increase in apoptosis has been reported upon miR-21-5p inhibition in L428 cells.…”

Section: Discussioncontrasting

confidence: 98%

MicroRNA High Throughput Loss-of-Function Screening Reveals an Oncogenic Role for miR-21-5p in Hodgkin Lymphoma

Yuan

Niu²,

Nolte

et al. 2018

Cell Physiol Biochem

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Background/Aims: Classical Hodgkin lymphoma (cHL) is among the most frequent lymphoma subtypes. The tumor cells originate from crippled germinal center (GC)-B cells that escaped from apoptosis. MicroRNAs (miRNAs) play important roles in B-cell maturation and aberrant expression of miRNAs contributes to the pathogenesis of cHL. Our aim was to identify oncogenic miRNAs relevant for growth of cHL using a high-throughput screening approach. Methods: A lentiviral pool of 63 miRNA inhibition constructs was used to identify miRNAs essential to cell growth in three cHL cell lines in duplicate. As a negative control we also infected cHL cell lines with a lentiviral barcoded empty vector pool consisting of 222 constructs. The abundance of individual constructs was followed over time by a next generation sequencing approach. The effect on growth was confirmed using individual GFP competition assays and on apoptosis using Annexin-V staining. Our previously published Argonaute 2 (Ago2) immunoprecipitation (IP) data were used to identify target genes relevant for cell growth / apoptosis. Luciferase assays and western blotting were performed to confirm targeting by miRNAs. Results: Four miRNA inhibition constructs, i.e. miR-449a-5p, miR-625-5p, let-7f-2-3p and miR-21-5p, showed a significant decrease in abundance in at least 4 of 6 infections. In contrast, none of the empty vector constructs showed a significant decrease in abundance in 3 or more of the 6 infections. The most abundantly expressed miRNA, i.e. miR-21-5p, showed significantly higher expression levels in cHL compared to GC-B cells. GFP competition assays confirmed the negative effect of miR-21-5p inhibition on HL cell growth. Annexin-V staining of cells infected with miR-21-5p inhibitor indicated a significant increase in apoptosis at day 7 and 9 after viral infection, consistent with the decrease in growth. Four miR-21-5p cell growth- and apoptosis-associated targets were AGO2-IP enriched in cHL cell lines and showed a significant decrease in expression in cHL cell lines in comparison to normal GC-B cells. For the two most abundantly expressed, i.e. BTG2 and PELI1, we confirmed targeting by miR-21-5p using luciferase assays and for PELI1 we also confirmed this at the protein level by western blotting. Conclusion: Using a miRNA loss-of-function high-throughput screen we identified four miRNAs with oncogenic effects in cHL and validated the results for the in cHL abundantly expressed miR-21-5p. MiR-21-5p is upregulated in cHL compared to GC-B cells and protects cHL cells from apoptosis possibly via targeting BTG2 and PELI1.

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Section: Discussioncontrasting

confidence: 98%

MicroRNA High Throughput Loss-of-Function Screening Reveals an Oncogenic Role for miR-21-5p in Hodgkin Lymphoma

Yuan

Niu²,

Nolte

et al. 2018

Cell Physiol Biochem

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“…Further studies will be performed to clarify this data. Van Eijndhoven et al confirmed our results in terms of high levels of EVs in patients with respect to controls [141]. Moreover, we and others reported that plasma EV miRNA levels, including miR155, miR127, miR21, and let7, reflect the presence of the tumor and could be a diagnostic tool for therapy response and relapse monitoring in HL [139,141].…”

Section: Clinical Potential Of Evs As Biomarkers and Therapeuticssupporting

confidence: 87%

Extracellular Vesicles in Hematological Malignancies: From Biology to Therapy

Caivano

Rocca

Laurenzana

et al. 2017

IJMS

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Extracellular vesicles (EVs) are a heterogeneous group of particles, between 15 nanometers and 10 microns in diameter, released by almost all cell types in physiological and pathological conditions, including tumors. EVs have recently emerged as particularly interesting informative vehicles, so that they could be considered a true “cell biopsy”. Indeed, EV cargo, including proteins, lipids, and nucleic acids, generally reflects the nature and status of the origin cells. In some cases, EVs are enriched of peculiar molecular cargo, thus suggesting at least a degree of specific cellular packaging. EVs are identified as important and critical players in intercellular communications in short and long distance interplays. Here, we examine the physiological role of EVs and their activity in cross-talk between bone marrow microenvironment and neoplastic cells in hematological malignancies (HMs). In these diseases, HM EVs can modify tumor and bone marrow microenvironment, making the latter “stronger” in supporting malignancy, inducing drug resistance, and suppressing the immune system. Moreover, EVs are abundant in biologic fluids and protect their molecular cargo against degradation. For these and other “natural” characteristics, EVs could be potential biomarkers in a context of HM liquid biopsy and therapeutic tools. These aspects will be also analyzed in this review.

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“…Serum, plasma, whole blood, nasopharyngeal brushing and frozen tumor biopsy samples were collected and processed for DNA and RNA isolation according to published protocols . Extracellular vesicles (EVs) were isolated from serum and plasma by SEC Sepharose CL‐2B column (GE Healthcare) as described previously . TRIZOL LS (Invitrogen) was used to isolate total RNA from 250 μL total serum or plasma according to the manufacturer's instruction.…”

Section: Methodsmentioning

confidence: 99%

Vesicle‐bound EBV‐BART13‐3p miRNA in circulation distinguishes nasopharyngeal from other head and neck cancer and asymptomatic EBV‐infections

Ramayanti

Verkuijlen

Novianti

et al. 2018

Intl Journal of Cancer

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Cell‐free microRNA (miRNA) in biofluids released by tumors in either protein or vesicle‐bound form, represent promising minimally‐invasive cancer biomarkers. However, a highly abundant non‐tumor background in human plasma and serum complicates the discovery and detection of tumor‐selective circulating miRNAs. We performed small RNA sequencing on serum and plasma RNA from Nasopharyngeal Carcinoma (NPC) patients. Collectively, Epstein Barr virus‐encoded miRNAs, more so than endogenous miRNAs, signify presence of NPC. However, RNAseq‐based EBV miRNA profiles differ between NPC patients, suggesting inter‐tumor heterogeneity or divergent secretory characteristics. We determined with sensitive qRT‐PCR assays that EBV miRNAs BART7‐3p, BART9‐3p and BART13‐3p are actively secreted by C666.1 NPC cells bound to extracellular vesicles (EVs) and soluble ribonucleoprotein complexes. Importantly, these miRNAs are expressed in all primary NPC tumor biopsies and readily detected in nasopharyngeal brushings from both early and late‐stage NPC patients. Increased levels of BART7‐3p, BART9‐3p and particularly BART13‐3p, distinguish NPC patient sera from healthy controls. Receiver operating characteristic curve analysis using sera from endemic NPC patients, other head and neck cancers and individuals with asymptomatic EBV‐infections reveals a superior diagnostic performance of EBV miRNAs over anti‐EBNA1 IgA serology and EBV‐DNA load (AUC 0.87–0.96 vs 0.86 and 0.66 respectively). The high specificity of circulating EBV‐BART13‐3p (97%) for NPC detection is in agreement with active secretion from NPC tumor cells. We conclude EV‐bound BART13‐3p in circulation is a promising, NPC‐selective, biomarker that should be considered as part of a screening strategy to identify NPC in endemic regions.

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Plasma vesicle miRNAs for therapy response monitoring in Hodgkin lymphoma patients

Cited by 143 publications

References 50 publications

MicroRNA High Throughput Loss-of-Function Screening Reveals an Oncogenic Role for miR-21-5p in Hodgkin Lymphoma

MicroRNA High Throughput Loss-of-Function Screening Reveals an Oncogenic Role for miR-21-5p in Hodgkin Lymphoma

Extracellular Vesicles in Hematological Malignancies: From Biology to Therapy

Vesicle‐bound EBV‐BART13‐3p miRNA in circulation distinguishes nasopharyngeal from other head and neck cancer and asymptomatic EBV‐infections

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