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Pre-analytical handling conditions and protein marker recovery from urine extracellular vesicles for bladder cancer diagnosis

Extracellular vesicles (EVs) contain a variety of biomolecules and provide information about the cells that produce them. EVs from cancer cells found in urine can be used as biomarkers to detect cancer, enabling early diagnosis and treatment. The potential of alpha-2-macroglobulin (A2M) and clusteri...

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Autores principales: Lee, Jisu, Kim, Eunha, Park, Joohee, Choi, Seokjoo, Lee, Myung-Shin, Park, Jinsung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10484439/
https://www.ncbi.nlm.nih.gov/pubmed/37676879
http://dx.doi.org/10.1371/journal.pone.0291198
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author Lee, Jisu
Kim, Eunha
Park, Joohee
Choi, Seokjoo
Lee, Myung-Shin
Park, Jinsung
author_facet Lee, Jisu
Kim, Eunha
Park, Joohee
Choi, Seokjoo
Lee, Myung-Shin
Park, Jinsung
author_sort Lee, Jisu
collection PubMed
description Extracellular vesicles (EVs) contain a variety of biomolecules and provide information about the cells that produce them. EVs from cancer cells found in urine can be used as biomarkers to detect cancer, enabling early diagnosis and treatment. The potential of alpha-2-macroglobulin (A2M) and clusterin (CLU) as novel diagnostic urinary EV (uEV) biomarkers for bladder cancer (BC) was demonstrated previously. To validate the diagnostic value of these proteins in uEVs in a large BC cohort, urine handling conditions before uEV isolation should be optimized during sample transportation from medical centers. In this study, we analyzed the uEV protein quantity, EV particle number, and uEV-A2M/CLU after urine storage at 20°C and 4°C for 0–6 days, each. A2M and CLU levels in uEVs were relatively stable when stored at 4°C for a maximum of three days and at 20°C for up to 24 h, with minimal impact on analysis results. Interestingly, pre-processing to remove debris and cells by centrifugation and filtration of urine did not show any beneficial effects on the preservation of protein biomarkers of uEVs during storage. Here, the importance of optimizing shipping conditions to minimize the impact of pre-analytical handling on the uEVs protein biomarkers was emphasized. These findings provide insights for the development of clinical protocols that use uEVs for diagnostic purposes.
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spelling pubmed-104844392023-09-08 Pre-analytical handling conditions and protein marker recovery from urine extracellular vesicles for bladder cancer diagnosis Lee, Jisu Kim, Eunha Park, Joohee Choi, Seokjoo Lee, Myung-Shin Park, Jinsung PLoS One Research Article Extracellular vesicles (EVs) contain a variety of biomolecules and provide information about the cells that produce them. EVs from cancer cells found in urine can be used as biomarkers to detect cancer, enabling early diagnosis and treatment. The potential of alpha-2-macroglobulin (A2M) and clusterin (CLU) as novel diagnostic urinary EV (uEV) biomarkers for bladder cancer (BC) was demonstrated previously. To validate the diagnostic value of these proteins in uEVs in a large BC cohort, urine handling conditions before uEV isolation should be optimized during sample transportation from medical centers. In this study, we analyzed the uEV protein quantity, EV particle number, and uEV-A2M/CLU after urine storage at 20°C and 4°C for 0–6 days, each. A2M and CLU levels in uEVs were relatively stable when stored at 4°C for a maximum of three days and at 20°C for up to 24 h, with minimal impact on analysis results. Interestingly, pre-processing to remove debris and cells by centrifugation and filtration of urine did not show any beneficial effects on the preservation of protein biomarkers of uEVs during storage. Here, the importance of optimizing shipping conditions to minimize the impact of pre-analytical handling on the uEVs protein biomarkers was emphasized. These findings provide insights for the development of clinical protocols that use uEVs for diagnostic purposes. Public Library of Science 2023-09-07 /pmc/articles/PMC10484439/ /pubmed/37676879 http://dx.doi.org/10.1371/journal.pone.0291198 Text en © 2023 Lee et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Lee, Jisu
Kim, Eunha
Park, Joohee
Choi, Seokjoo
Lee, Myung-Shin
Park, Jinsung
Pre-analytical handling conditions and protein marker recovery from urine extracellular vesicles for bladder cancer diagnosis
title Pre-analytical handling conditions and protein marker recovery from urine extracellular vesicles for bladder cancer diagnosis
title_full Pre-analytical handling conditions and protein marker recovery from urine extracellular vesicles for bladder cancer diagnosis
title_fullStr Pre-analytical handling conditions and protein marker recovery from urine extracellular vesicles for bladder cancer diagnosis
title_full_unstemmed Pre-analytical handling conditions and protein marker recovery from urine extracellular vesicles for bladder cancer diagnosis
title_short Pre-analytical handling conditions and protein marker recovery from urine extracellular vesicles for bladder cancer diagnosis
title_sort pre-analytical handling conditions and protein marker recovery from urine extracellular vesicles for bladder cancer diagnosis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10484439/
https://www.ncbi.nlm.nih.gov/pubmed/37676879
http://dx.doi.org/10.1371/journal.pone.0291198
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