Impact of Extracellular Vesicle Isolation Methods on Downstream miRNA Analysis in Semen: A Comparative Study

Autor: Lluís Bassas, Manel Castells, F. Vigués, Olga López-Rodrigo, Alexandre de la Fuente, Carolina Herrero, Marina Mercadal, Sara Larriba
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Male
0301 basic medicine
diagnosis
extracellular vesicle isolation methods
lcsh:Chemistry
Prostate cancer
0302 clinical medicine
Vesícules seminals
lcsh:QH301-705.5
Spectroscopy
Seminal vesicles
Biochemical markers
semen
General Medicine
Extracellular vesicle
Middle Aged
prostate cancer
Computer Science Applications
Prostate-specific antigen
Biochemistry
030220 oncology & carcinogenesis
Marcadors bioquímics
Biomarker (medicine)
biomarker
extracellular vesicles
microvesicles
Adult
Semen
exosomes
Biology
Cell Fractionation
Exosome
Catalysis
Article
Inorganic Chemistry
03 medical and health sciences
microRNA
Biomarkers
Tumor
medicine
Humans
Physical and Theoretical Chemistry
Molecular Biology
Aged
Càncer de pròstata
Organic Chemistry
Liquid Biopsy
Prostatic Neoplasms
medicine.disease
Microvesicles
MicroRNAs
030104 developmental biology
lcsh:Biology (General)
lcsh:QD1-999
extracellular vesicle miRNA analysis
Zdroj: International Journal of Molecular Sciences
Dipòsit Digital de la UB
Universidad de Barcelona
Volume 21
Issue 17
International Journal of Molecular Sciences, Vol 21, Iss 5949, p 5949 (2020)
ISSN: 1422-0067
Popis: 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.
Databáze: OpenAIRE
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