A novel isolation method for spontaneously released extracellular vesicles from brain tissue and its implications for stress-driven brain pathology

Cell Communication and Signaling - Tập 21 Số 1
Patrícia Gomes1, Cristian Bodo2, Carlos Nogueras‐Ortiz3, Martina Samiotaki4, Minghao Chen5, Carina Soares‐Cunha1, Joana M Silva6, Bárbara Coimbra1, George Stamatakis4, Liliana Santos7, George Panayotou4, Foteini Tzouanou8, Clarissa L. Waites9, Christos Gatsogiannis5, Nuno Sousa6, Dimitrios Kapogiannis3, Bruno Costa-Silva2, Ioannis Sotiropoulos6
1Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal
2Systems Oncology Group, Champalimaud Research, Champalimaud Centre for the Unknown, Av. Brasília, 1400-038, Lisbon, Portugal
3Laboratory of Clinical Investigation, Intramural Research Program, National Institute on Aging, NIH, Baltimore, MD, USA
4Institute for Bioinnovation, Biomedical Sciences Research Center “Alexander Fleming”, 16672, Vari, Attica, Greece
5Center for Soft Nanoscience and Institute of Medical Physics and Biophysics, University of Muenster, 48149, Münster, Germany
6ICVS/3B’s - PT Government Associate Laboratory, Braga/Guimarães, Portugal
7ICVS/3B’s – PT Government Associate Laboratory, Braga/Guimarães, Portugal
8Institute of Biosciences and Applications, NCSR Demokritos, Athens, Greece
9Department of Pathology and Cell Biology, Taub Institute for Research on Alzheimer’s Disease and the Aging Brain, Columbia University Medical Center, New York, NY, USA

Tóm tắt

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.

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Cell Communication and Signaling

Tập 21 Số 1

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