Plasma-derived extracellular vesicles contain mutant SOD1 in hSOD1G93A transgenic swine

Background: Two goals of amyotrophic lateral sclerosis (ALS) research are (a) validation of new experimental models and (b) identification of diagnostic biomarkers, in order to speed up the diagnosis, to monitor its progression and to assess whether a new therapy may be effective. Extracellular vesi...

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Bibliographic Details
Published inJournal of extracellular vesicles Vol. 7; p. 158
Main Authors Berrone, Elena, Crociara, Paola, Lo Faro, Monica, Costassa, Elena Vallino, Favole, Alessandra, Deregibus, Maria Chiara, Camussi, Giovanni, Galli, Cesare, Duchi, Roberto, Chiò, Adriano, Calvo, Andrea, Casale, Federico, Fuda, Giuseppe, De Marco, Giovanni, Casalone, Cristina, Corona, Cristiano
Format Journal Article
LanguageEnglish
Published Abingdon John Wiley & Sons, Inc 01.01.2018
Subjects
Amyotrophic lateral sclerosis
Animal models
Biomarkers
CD63 antigen
Diagnosis
Exosomes
Extracellular vesicles
Flow cytometry
Immunoprecipitation
Phenotypes
Proteins
Superoxide dismutase
Western blotting
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Summary:Background: Two goals of amyotrophic lateral sclerosis (ALS) research are (a) validation of new experimental models and (b) identification of diagnostic biomarkers, in order to speed up the diagnosis, to monitor its progression and to assess whether a new therapy may be effective. Extracellular vesicles (EVs) and their content may be a reliable clinical biomarker for ALS, as they have yet been used for the diagnosis and prognosis of various diseases. In this context, we developed an hSOD1G93A transgenic swine characterized by a long preclinical and clinical phase in order to clarify certain ALS etiopathogenetic aspects. In particular, EVs characterization in this animal model could elucidate their role in relation to key elements of the disease process. Therefore, this study aimed at evaluating hSOD1 protein into EVs isolated from hSOD1G93A transgenic swine plasma. Methods: EVs were isolated from plasma of hSOD1G93A and wild-type (WT) pigs by a modified precipitation method. EVs were characterized by Tunable Resistive Pulse Sensing, flow cytometry (Cytoflex Beckman) and Western blotting (WB). After immunoprecipitation of lysed EVs, WT and hSOD1 protein detection was performed by WB. Results: Phenotype characterization confirmed that the majority of EVs were exosomes (particle diameter mean: 119 nm, %: 52,29) expressing the typical exosome markers (CD63, TSG101, Flotillin 1, Alix). As regard SOD1 analysis, the hSOD1 protein with the G93A mutation was found only in plasmaderived exosomes of the transgenic swine model, but not in WT. Summary/Conclusion: These results showed that the majority of EVs derived from hSOD1G93A swine model are exosomes able to carry hSOD1G93A protein. Therefore, parallel investigations of exosomes on ALS patients and hSOD1G93A swine model could clarify their role in the pathogenesis of the disease and could represent a new diagnostic and therapeutic strategy.
ISSN:2001-3078