A novel isolation method for spontaneously released extracellular vesicles from brain tissue and its implications for stress-driven brain pathology
Abstract Background Extracellular vesicles (EVs), including small EVs (sEVs) such as exosomes, exhibit great potential for the diagnosis and treatment of brain disorders, representing a valuable tool for precision medicine. The latter demands high-quality human biospecimens, especially in complex di...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-02-01
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| Series: | Cell Communication and Signaling |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12964-023-01045-z |
| _version_ | 1797863999501500416 |
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| author | Patrícia A. Gomes Cristian Bodo Carlos Nogueras-Ortiz Martina Samiotaki Minghao Chen Carina Soares-Cunha Joana M. Silva Bárbara Coimbra George Stamatakis Liliana Santos George Panayotou Foteini Tzouanou Clarissa L. Waites Christos Gatsogiannis Nuno Sousa Dimitrios Kapogiannis Bruno Costa-Silva Ioannis Sotiropoulos |
| author_facet | Patrícia A. Gomes Cristian Bodo Carlos Nogueras-Ortiz Martina Samiotaki Minghao Chen Carina Soares-Cunha Joana M. Silva Bárbara Coimbra George Stamatakis Liliana Santos George Panayotou Foteini Tzouanou Clarissa L. Waites Christos Gatsogiannis Nuno Sousa Dimitrios Kapogiannis Bruno Costa-Silva Ioannis Sotiropoulos |
| author_sort | Patrícia A. Gomes |
| collection | DOAJ |
| description | Abstract Background Extracellular vesicles (EVs), including small EVs (sEVs) such as exosomes, exhibit great potential for the diagnosis and treatment of brain disorders, representing a valuable tool for precision medicine. The latter demands high-quality human biospecimens, especially in complex disorders in which pathological and specimen heterogeneity, as well as diverse individual clinical profile, often complicate the development of precision therapeutic schemes and patient-tailored treatments. Thus, the collection and characterization of physiologically relevant sEVs are of the utmost importance. However, standard brain EV isolation approaches rely on tissue dissociation, which can contaminate EV fractions with intracellular vesicles. Methods Based on multiscale analytical platforms such as cryo-EM, label-free proteomics, advanced flow cytometry, and ExoView analyses, we compared and characterized the EV fraction isolated with this novel method with a classical digestion-based EV isolation procedure. Moreover, EV biogenesis was pharmacologically manipulated with either GW4869 or picrotoxin to assess the validity of the spontaneous-release method, while the injection of labelled-EVs into the mouse brain further supported the integrity of the isolated vesicles. Results We hereby present an efficient purification method that captures a sEV-enriched population spontaneously released by mouse and human brain tissue. In addition, we tested the significance of the release method under conditions where biogenesis/secretion of sEVs was pharmacologically manipulated, as well as under animals’ exposure to chronic stress, a clinically relevant precipitant of brain pathologies, such as depression and Alzheimer’s disease. Our findings show that the released method monitors the drug-evoked inhibition or enhancement of sEVs secretion while chronic stress induces the secretion of brain exosomes accompanied by memory loss and mood deficits suggesting a potential role of sEVs in the brain response to stress and related stress-driven brain pathology. Conclusions Overall, the spontaneous release method of sEV yield may contribute to the characterization and biomarker profile of physiologically relevant brain-derived sEVs in brain function and pathology. Video Abstract |
| first_indexed | 2024-04-09T22:45:35Z |
| format | Article |
| id | doaj.art-1b9f0ce8293249e8b5b641bda464ebec |
| institution | Directory Open Access Journal |
| issn | 1478-811X |
| language | English |
| last_indexed | 2024-04-09T22:45:35Z |
| publishDate | 2023-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell Communication and Signaling |
| spelling | doaj.art-1b9f0ce8293249e8b5b641bda464ebec2023-03-22T11:53:00ZengBMCCell Communication and Signaling1478-811X2023-02-0121111610.1186/s12964-023-01045-zA novel isolation method for spontaneously released extracellular vesicles from brain tissue and its implications for stress-driven brain pathologyPatrícia A. Gomes0Cristian Bodo1Carlos Nogueras-Ortiz2Martina Samiotaki3Minghao Chen4Carina Soares-Cunha5Joana M. Silva6Bárbara Coimbra7George Stamatakis8Liliana Santos9George Panayotou10Foteini Tzouanou11Clarissa L. Waites12Christos Gatsogiannis13Nuno Sousa14Dimitrios Kapogiannis15Bruno Costa-Silva16Ioannis Sotiropoulos17Life and Health Sciences Research Institute (ICVS), School of Medicine, University of MinhoSystems Oncology Group, Champalimaud Research, Champalimaud Centre for the UnknownLaboratory of Clinical Investigation, Intramural Research Program, National Institute on Aging, NIHInstitute for Bioinnovation, Biomedical Sciences Research Center “Alexander Fleming”Center for Soft Nanoscience and Institute of Medical Physics and Biophysics, University of MuensterLife and Health Sciences Research Institute (ICVS), School of Medicine, University of MinhoLife and Health Sciences Research Institute (ICVS), School of Medicine, University of MinhoLife and Health Sciences Research Institute (ICVS), School of Medicine, University of MinhoInstitute for Bioinnovation, Biomedical Sciences Research Center “Alexander Fleming”Life and Health Sciences Research Institute (ICVS), School of Medicine, University of MinhoInstitute for Bioinnovation, Biomedical Sciences Research Center “Alexander Fleming”Institute of Biosciences and Applications NCSR “Demokritos”Department of Pathology and Cell Biology, Taub Institute for Research on Alzheimer’s Disease and the Aging Brain, Columbia University Medical CenterCenter for Soft Nanoscience and Institute of Medical Physics and Biophysics, University of MuensterLife and Health Sciences Research Institute (ICVS), School of Medicine, University of MinhoLaboratory of Clinical Investigation, Intramural Research Program, National Institute on Aging, NIHSystems Oncology Group, Champalimaud Research, Champalimaud Centre for the UnknownLife and Health Sciences Research Institute (ICVS), School of Medicine, University of MinhoAbstract Background Extracellular vesicles (EVs), including small EVs (sEVs) such as exosomes, exhibit great potential for the diagnosis and treatment of brain disorders, representing a valuable tool for precision medicine. The latter demands high-quality human biospecimens, especially in complex disorders in which pathological and specimen heterogeneity, as well as diverse individual clinical profile, often complicate the development of precision therapeutic schemes and patient-tailored treatments. Thus, the collection and characterization of physiologically relevant sEVs are of the utmost importance. However, standard brain EV isolation approaches rely on tissue dissociation, which can contaminate EV fractions with intracellular vesicles. Methods Based on multiscale analytical platforms such as cryo-EM, label-free proteomics, advanced flow cytometry, and ExoView analyses, we compared and characterized the EV fraction isolated with this novel method with a classical digestion-based EV isolation procedure. Moreover, EV biogenesis was pharmacologically manipulated with either GW4869 or picrotoxin to assess the validity of the spontaneous-release method, while the injection of labelled-EVs into the mouse brain further supported the integrity of the isolated vesicles. Results We hereby present an efficient purification method that captures a sEV-enriched population spontaneously released by mouse and human brain tissue. In addition, we tested the significance of the release method under conditions where biogenesis/secretion of sEVs was pharmacologically manipulated, as well as under animals’ exposure to chronic stress, a clinically relevant precipitant of brain pathologies, such as depression and Alzheimer’s disease. Our findings show that the released method monitors the drug-evoked inhibition or enhancement of sEVs secretion while chronic stress induces the secretion of brain exosomes accompanied by memory loss and mood deficits suggesting a potential role of sEVs in the brain response to stress and related stress-driven brain pathology. Conclusions Overall, the spontaneous release method of sEV yield may contribute to the characterization and biomarker profile of physiologically relevant brain-derived sEVs in brain function and pathology. Video Abstracthttps://doi.org/10.1186/s12964-023-01045-zExtracellular vesiclesBrainExosomesHumanMouseSpontaneous release |
| spellingShingle | Patrícia A. Gomes Cristian Bodo Carlos Nogueras-Ortiz Martina Samiotaki Minghao Chen Carina Soares-Cunha Joana M. Silva Bárbara Coimbra George Stamatakis Liliana Santos George Panayotou Foteini Tzouanou Clarissa L. Waites Christos Gatsogiannis Nuno Sousa Dimitrios Kapogiannis Bruno Costa-Silva Ioannis Sotiropoulos A novel isolation method for spontaneously released extracellular vesicles from brain tissue and its implications for stress-driven brain pathology Cell Communication and Signaling Extracellular vesicles Brain Exosomes Human Mouse Spontaneous release |
| title | A novel isolation method for spontaneously released extracellular vesicles from brain tissue and its implications for stress-driven brain pathology |
| title_full | A novel isolation method for spontaneously released extracellular vesicles from brain tissue and its implications for stress-driven brain pathology |
| title_fullStr | A novel isolation method for spontaneously released extracellular vesicles from brain tissue and its implications for stress-driven brain pathology |
| title_full_unstemmed | A novel isolation method for spontaneously released extracellular vesicles from brain tissue and its implications for stress-driven brain pathology |
| title_short | A novel isolation method for spontaneously released extracellular vesicles from brain tissue and its implications for stress-driven brain pathology |
| title_sort | novel isolation method for spontaneously released extracellular vesicles from brain tissue and its implications for stress driven brain pathology |
| topic | Extracellular vesicles Brain Exosomes Human Mouse Spontaneous release |
| url | https://doi.org/10.1186/s12964-023-01045-z |
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