A method of separating extracellular vesicles from blood shows potential clinical translation, and reveals extracellular vesicle cargo gremlin-1 as a diagnostic biomarker
Extracellular vesicles (EVs) have potential as minimally invasive biomarkers. However, the methods most commonly used for EV retrieval rely on ultracentrifugation, are time-consuming, and unrealistic to translate to standard-of-care. We sought a method suitable for EV separation from blood that coul...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-01-01
|
| Series: | Translational Oncology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1936523321002655 |
| _version_ | 1819179131474018304 |
|---|---|
| author | Niamh McNamee Róisín Daly John Crown Lorraine O'Driscoll |
| author_facet | Niamh McNamee Róisín Daly John Crown Lorraine O'Driscoll |
| author_sort | Niamh McNamee |
| collection | DOAJ |
| description | Extracellular vesicles (EVs) have potential as minimally invasive biomarkers. However, the methods most commonly used for EV retrieval rely on ultracentrifugation, are time-consuming, and unrealistic to translate to standard-of-care. We sought a method suitable for EV separation from blood that could be used in patient care. Sera from breast cancer patients and age-matched controls (n = 27 patients; n = 36 controls) were analysed to compare 6 proposed EV separation methods. The EVs were then characterised on 8 parameters. The selected method was subsequently applied to independent cohorts of sera (n = 20 patients; n = 20 controls), as proof-of-principle, investigating EVs’ gremlin-1 cargo. Three independent runs with each method were very reproducible, within each given method. All isolates contained EVs, although they varied in quantity and purity. Methods that require ultracentrifugation were not superior for low volumes of sera typically available in routine standard-of-care. A CD63/CD81/CD9-coated immunobead-based method was most suitable based on EV markers' detection and minimal albumin and lipoprotein contamination. Applying this method to independent sera cohorts, EVs and their gremlin-1 cargo were at significantly higher amounts for breast cancer patients compared to controls. In conclusion, CD63/CD81/CD9-coated immunobeads may enable clinical utility of blood-based EVs as biomarkers. |
| first_indexed | 2024-12-22T21:53:34Z |
| format | Article |
| id | doaj.art-a26c367adee14f9fa1c6bada14223445 |
| institution | Directory Open Access Journal |
| issn | 1936-5233 |
| language | English |
| last_indexed | 2024-12-22T21:53:34Z |
| publishDate | 2022-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Translational Oncology |
| spelling | doaj.art-a26c367adee14f9fa1c6bada142234452022-12-21T18:11:19ZengElsevierTranslational Oncology1936-52332022-01-01151101274A method of separating extracellular vesicles from blood shows potential clinical translation, and reveals extracellular vesicle cargo gremlin-1 as a diagnostic biomarkerNiamh McNamee0Róisín Daly1John Crown2Lorraine O'Driscoll3School of Pharmacy and Pharmaceutical Sciences, Panoz Institute, Trinity College Dublin, Dublin, Ireland; Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland; Trinity St. James's Cancer Institute, Trinity College Dublin, Dublin, IrelandSchool of Pharmacy and Pharmaceutical Sciences, Panoz Institute, Trinity College Dublin, Dublin, Ireland; Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland; Trinity St. James's Cancer Institute, Trinity College Dublin, Dublin, IrelandDepartment of Medical Oncology, St. Vincent's University Hospital (SVUH), Dublin, IrelandSchool of Pharmacy and Pharmaceutical Sciences, Panoz Institute, Trinity College Dublin, Dublin, Ireland; Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland; Trinity St. James's Cancer Institute, Trinity College Dublin, Dublin, Ireland; Corresponding author at: School of Pharmacy and Pharmaceutical Sciences, Trinity College Dublin, Trinity Biomedical Sciences Institute, Dublin, Ireland.Extracellular vesicles (EVs) have potential as minimally invasive biomarkers. However, the methods most commonly used for EV retrieval rely on ultracentrifugation, are time-consuming, and unrealistic to translate to standard-of-care. We sought a method suitable for EV separation from blood that could be used in patient care. Sera from breast cancer patients and age-matched controls (n = 27 patients; n = 36 controls) were analysed to compare 6 proposed EV separation methods. The EVs were then characterised on 8 parameters. The selected method was subsequently applied to independent cohorts of sera (n = 20 patients; n = 20 controls), as proof-of-principle, investigating EVs’ gremlin-1 cargo. Three independent runs with each method were very reproducible, within each given method. All isolates contained EVs, although they varied in quantity and purity. Methods that require ultracentrifugation were not superior for low volumes of sera typically available in routine standard-of-care. A CD63/CD81/CD9-coated immunobead-based method was most suitable based on EV markers' detection and minimal albumin and lipoprotein contamination. Applying this method to independent sera cohorts, EVs and their gremlin-1 cargo were at significantly higher amounts for breast cancer patients compared to controls. In conclusion, CD63/CD81/CD9-coated immunobeads may enable clinical utility of blood-based EVs as biomarkers.http://www.sciencedirect.com/science/article/pii/S1936523321002655Extracellular vesiclesbreast cancerClinical utilitystandard-of-careGremlin-1 |
| spellingShingle | Niamh McNamee Róisín Daly John Crown Lorraine O'Driscoll A method of separating extracellular vesicles from blood shows potential clinical translation, and reveals extracellular vesicle cargo gremlin-1 as a diagnostic biomarker Translational Oncology Extracellular vesicles breast cancer Clinical utility standard-of-care Gremlin-1 |
| title | A method of separating extracellular vesicles from blood shows potential clinical translation, and reveals extracellular vesicle cargo gremlin-1 as a diagnostic biomarker |
| title_full | A method of separating extracellular vesicles from blood shows potential clinical translation, and reveals extracellular vesicle cargo gremlin-1 as a diagnostic biomarker |
| title_fullStr | A method of separating extracellular vesicles from blood shows potential clinical translation, and reveals extracellular vesicle cargo gremlin-1 as a diagnostic biomarker |
| title_full_unstemmed | A method of separating extracellular vesicles from blood shows potential clinical translation, and reveals extracellular vesicle cargo gremlin-1 as a diagnostic biomarker |
| title_short | A method of separating extracellular vesicles from blood shows potential clinical translation, and reveals extracellular vesicle cargo gremlin-1 as a diagnostic biomarker |
| title_sort | method of separating extracellular vesicles from blood shows potential clinical translation and reveals extracellular vesicle cargo gremlin 1 as a diagnostic biomarker |
| topic | Extracellular vesicles breast cancer Clinical utility standard-of-care Gremlin-1 |
| url | http://www.sciencedirect.com/science/article/pii/S1936523321002655 |
| work_keys_str_mv | AT niamhmcnamee amethodofseparatingextracellularvesiclesfrombloodshowspotentialclinicaltranslationandrevealsextracellularvesiclecargogremlin1asadiagnosticbiomarker AT roisindaly amethodofseparatingextracellularvesiclesfrombloodshowspotentialclinicaltranslationandrevealsextracellularvesiclecargogremlin1asadiagnosticbiomarker AT johncrown amethodofseparatingextracellularvesiclesfrombloodshowspotentialclinicaltranslationandrevealsextracellularvesiclecargogremlin1asadiagnosticbiomarker AT lorraineodriscoll amethodofseparatingextracellularvesiclesfrombloodshowspotentialclinicaltranslationandrevealsextracellularvesiclecargogremlin1asadiagnosticbiomarker AT niamhmcnamee methodofseparatingextracellularvesiclesfrombloodshowspotentialclinicaltranslationandrevealsextracellularvesiclecargogremlin1asadiagnosticbiomarker AT roisindaly methodofseparatingextracellularvesiclesfrombloodshowspotentialclinicaltranslationandrevealsextracellularvesiclecargogremlin1asadiagnosticbiomarker AT johncrown methodofseparatingextracellularvesiclesfrombloodshowspotentialclinicaltranslationandrevealsextracellularvesiclecargogremlin1asadiagnosticbiomarker AT lorraineodriscoll methodofseparatingextracellularvesiclesfrombloodshowspotentialclinicaltranslationandrevealsextracellularvesiclecargogremlin1asadiagnosticbiomarker |