![]() Other isolation methods include size exclusion chromatography, density gradient centrifugation, filtration, and polymer-based precipitation. ![]() However, this method co-isolates high-density lipoproteins, is quite cumbersome, may result in clumping of EVs, and can lead to damage of EVs and sample loss. Ultracentrifugation has long been considered the preferred method for sEV isolation. Further studies are needed to clarify whether EVs actually contain DNA, and if so, the biomarker potential of such DNA. Another hypothesis is that DNA is not associated with circulating EVs but rather is carried by multivesicular endosomes and released into the extracellular space upon fusion with the plasma membrane. It has been proposed that evDNA is bound outside the vesicles, with only a minute portion present inside. However, several studies have reported conflicting results. Recent reports also have implied that the tumor fraction of EV-derived DNA (evDNA) might be higher than the tumor fraction in comparative cell-free DNA (cfDNA) in patients with cancer, making EVs a potential source of tumor-derived DNA in the circulation. Subgroups of EVs are reported to contain nucleic acids, and the discovery of DNA inside tumor-derived EVs has spawned numerous publications investigating its potential as a biomarker in cancer. EVs can be broadly categorized into three main classes based on size and the mode of biogenesis: a) exosomes, which are small (s)EVs (≤200 nm) shed by the fusion of multivesicular bodies to the plasma membrane b) microvesicles or microparticles, which are large (l)EVs (>200 nm) shed by outward budding of the plasma membrane and c) apoptotic bodies, which are released from fragments of apoptotic cells. They transfer molecules to other cells to influence recipient cell function and can vary in different features such as size, biogenesis, function, and cargo. ![]() The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have declared that no competing interests exist.Įxtracellular vesicles (EVs) are membrane-enclosed particles secreted by most cell types. received funding from the Folke Hermansen Foundation, the Western Norway Regional Health Authority ( and the Norwegian Cancer Society ( The project was part of a strategic program called “Personalized medicine-biomarkers and clinical studies,” supported by the Western Norway Regional Health Authority. received funding from the Folke Hermansen Foundation ( and the Western Norway Regional Health Authority ( O.N. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the paper and its Supporting Information files.įunding: M.L. Received: Accepted: SeptemPublished: September 14, 2023Ĭopyright: © 2023 Lapin et al. PLoS ONE 18(9):Įditor: Kira Astakhova, Danmarks Tekniske Universitet, DENMARK (2023) Extracellular vesicles as a potential source of tumor-derived DNA in advanced pancreatic cancer. In conclusion, our results suggest that quantification of evDNA, as a source of tumor-derived DNA, does not add information to that obtained with cfDNA, at least not in patients with advanced pancreatic cancer.Ĭitation: Lapin M, Tjensvoll K, Nedrebø K, Taksdal E, Janssen H, Gilje B, et al. ![]() Furthermore, the fraction of tumor-derived DNA in EVs was similar to that found in cfDNA. Most of the detected evDNA was also located on the outside of the vesicles. Our results indicated that the DNA content of EVs was significantly less than the cfDNA content isolated from the same plasma volume ( p < 0.001). To assess whether methodology affected the results, we isolated EVs using four different methods for small EV isolation and differential centrifugation for isolating large EVs. evDNA from both DNase-treated and untreated EV samples was analyzed to determine whether the DNA was primarily located internally or outside (surface-bound) the EVs. Here, we evaluated EVs as a potential source of tumor-derived DNA in patients with advanced pancreatic cancer. Several studies have highlighted the potential of EV-derived DNA (evDNA) as a circulating biomarker, even demonstrating that evDNA can outperform cell-free DNA (cfDNA) in terms of sensitivity. Tumor-derived extracellular vesicles (EVs) are reported to contain nucleic acids, including DNA.
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