Irreversible alteration of extracellular vesicle and cell-free messenger RNA profiles in human plasma associated with blood processing and storage.

Autor: Kim HJ; Cancer Early Detection Advanced Research Center, Knight Cancer Institute (CEDAR), Oregon Health and Science University, 2720 SW Moody Ave, KR-CEDR, Portland, OR, 97201, USA.; Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR, USA., Rames MJ; Cancer Early Detection Advanced Research Center, Knight Cancer Institute (CEDAR), Oregon Health and Science University, 2720 SW Moody Ave, KR-CEDR, Portland, OR, 97201, USA.; Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR, USA., Tassi Yunga S; Cancer Early Detection Advanced Research Center, Knight Cancer Institute (CEDAR), Oregon Health and Science University, 2720 SW Moody Ave, KR-CEDR, Portland, OR, 97201, USA.; Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR, USA., Armstrong R; Cancer Early Detection Advanced Research Center, Knight Cancer Institute (CEDAR), Oregon Health and Science University, 2720 SW Moody Ave, KR-CEDR, Portland, OR, 97201, USA., Morita M; Department of Pathology, Oregon Health and Science University, Portland, OR, USA., Ngo ATP; Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR, USA., McCarty OJT; Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR, USA., Civitci F; Cancer Early Detection Advanced Research Center, Knight Cancer Institute (CEDAR), Oregon Health and Science University, 2720 SW Moody Ave, KR-CEDR, Portland, OR, 97201, USA., Morgan TK; Cancer Early Detection Advanced Research Center, Knight Cancer Institute (CEDAR), Oregon Health and Science University, 2720 SW Moody Ave, KR-CEDR, Portland, OR, 97201, USA.; Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR, USA.; Department of Pathology, Oregon Health and Science University, Portland, OR, USA., Ngo TTM; Cancer Early Detection Advanced Research Center, Knight Cancer Institute (CEDAR), Oregon Health and Science University, 2720 SW Moody Ave, KR-CEDR, Portland, OR, 97201, USA. ngth@ohsu.edu.; Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR, USA. ngth@ohsu.edu.; Department of Molecular and Medical Genetics, Oregon Health and Science University, Portland, OR, USA. ngth@ohsu.edu.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2022 Feb 08; Vol. 12 (1), pp. 2099. Date of Electronic Publication: 2022 Feb 08.
DOI: 10.1038/s41598-022-06088-9
Abstrakt: The discovery and utility of clinically relevant circulating biomarkers depend on standardized methods that minimize preanalytical errors. Despite growing interest in studying extracellular vesicles (EVs) and cell-free messenger RNA (cf-mRNA) as potential biomarkers, how blood processing and freeze/thaw impacts the profiles of these analytes in plasma was not thoroughly understood. We utilized flow cytometric analysis to examine the effect of differential centrifugation and a freeze/thaw cycle on EV profiles. Utilizing flow cytometry postacquisition analysis software (FCMpass) to calibrate light scattering and fluorescence, we revealed how differential centrifugation and post-freeze/thaw processing removes and retains EV subpopulations. Additionally, cf-mRNA levels measured by RT-qPCR profiles from a panel of housekeeping, platelet, and tissue-specific genes were preferentially affected by differential centrifugation and post-freeze/thaw processing. Critically, freezing plasma containing residual platelets yielded irreversible ex vivo generation of EV subpopulations and cf-mRNA transcripts, which were not removable by additional processing after freeze/thaw. Our findings suggest the importance of minimizing confounding variation attributed to plasma processing and platelet contamination.
(© 2022. The Author(s).)
Databáze: MEDLINE
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