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

• Published in: Journal 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
• Language: English
• 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

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.