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Impact of Extracellular Vesicle Isolation Methods on Downstream miRNA Analysis in Semen

An article by Sara Larriba of the Human Molecular Genetics Group of Bellvitge Biomedical Research Institute (IDIBELL), recently published by the International Journal of Molecular Science. entitled “Impact of Extracellular Vesicle Isolation Methods on Downstream miRNA Analysis in Semen: A Comparative Study“, acknowledges NANBIOSIS ICTS for its participation in the research results. The nanoparticle tracking analysis was performed by the ICTS NANBIOSIS U6 Biomaterial Processing and Nanostructuring Unit.

Abstract:

Seminal plasma (SP) contains a unique concentration of miRNA, mostly contained in small extracellular vesicles (sEVs) such as exosomes, some of which could be clinically useful for diagnosis and/or prognosis of urogenital diseases such as prostate cancer (PCa). We optimized several exosome-EV isolation technologies for their use in semen, evaluating EV purifying effectiveness and impact on the downstream analysis of miRNAs against results from the standard ultracentrifugation (UC) method to implement the use of SP sEV_miRNAs as noninvasive biomarkers for PCa. Our results evidenced that commercial kits designed to isolate exosomes/EVs from blood or urine are mostly applicable to SP, but showed quantitative and qualitative variability between them. ExoGAG 3500× g and the miRCURY Cell/Urine/CSF 1500× g methods resulted as equivalent alternative procedures to UC for isolating exosomes/sEVs from semen for nanoparticle characteristics and quality of RNA contained in vesicles. Additionally, the expression profile of the altered semen sEV-miRNAs in PCa varies depending on the EV isolation method applied. This is possibly due to different extraction techniques yielding different proportions of sEV subtypes. This is evidence that the exosome-EV isolation method has a significant impact on the analysis of the miRNAs contained within, with important consequences for their use as clinical biomarkers. Therefore, miRNA analysis results for EVs cannot be directly extrapolated between different EV isolation methods until clear markers for delineation between microvesicles and exosomes are established. However, EV extraction methodology affects combined models (semen exosome miRNA signatures plus blood Prostate specific antigen (PSA) concentration for PCa diagnosis) less; specifically our previously described (miR-142-3p + miR-142-5p + miR-223-3p + PSA) model functions as molecular marker from EVs from any of the three isolation methods, potentially improving the efficiency of PSA PCa diagnosis..

Article:

Impact of Extracellular Vesicle Isolation Methods on Downstream miRNA Analysis in Semen: A Comparative Study. Marina Mercadal, Carolina Herrero, Olga López-Rodrigo, Manel Castells, Alexandre de la Fuente, Francesc Vigués, Lluís Bassas and Sara Larriba. Int. J. Mol. Sci. 2020, 21, 5949; doi:10.3390/ijms21175949

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Biomarkers in semen to diagnose prostate cancer

Sara Larriba of the Human Molecular Genetics Group of Bellvitge Biomedical Research Institute (IDIBELL) has informed NANBIOSIS about a recent publication mentioning NANBIOSIS in the Acknowledgements for its participation in the results of their research. (The nanoparticle tracking analysis was performed by the ICTS NANBIOSIS U6 Biomaterial Processing and Nanostructuring Unit) The article has been published by the journal Scientific Reports of Nature Research.

The prediction of PCa in the early stage of the disease is one of the most important objectives in male urology. A significant decrease in deaths due to PCa has been associated with the use of serum PSA test recent years. However, the PSA test still has serious limitations and often gives false positives that lead to many unnecessary biopsies of benign disease. Therefore, researchers from the Human Molecular Genetics Group of Bellvitge Biomedical Research Institute (IDIBELL) decided to evaluate semen as a source of prostate cancer biomarkers, and studying extracellular miRNAs, which are present within semen in extraordinary concentrations since some of the Some of these extracellular miRNAs are specific to the prostate gland and, in addition, there is already research showing that extracellular miRNAs can reflect altered patterns of miRNA expression in prostate tumor tissue. The study conducted allowed scientists to discover a distinctive miRNA expression pattern in exosomal semen samples obtained from men with prostate cancer compared with that found in exosomal semen samples taken from healthy men. The next step would be to conduct more prospective studies in larger patient cohorts before this miRNA-based biomarker can be adopted in daily clinical practice.

Article of reference:

Semen miRNAs Contained in Exosomes as Non-Invasive Biomarkers for Prostate Cancer Diagnosis, Maria Barceló, Manel Castells, Lluís Bassas, Francesc Vigués2 & Sara Larriba. Scientific Reports volume 9, 24 Sept 2019.

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