The Anti-Apoptotic Effect of ASC-Exosomes in an In Vitro ALS Model and Their Proteomic Analysis

Abstract

Stem cell therapy represents a promising approach in the treatment of several neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS). The beneficial effect of stem cells is exerted by paracrine mediators, as exosomes, suggesting a possible potential use of these extracellular vesicles as non-cell based therapy. We demonstrated that exosomes isolated from adipose stem cells (ASC) display a neuroprotective role in an in vitro model of ALS. Moreover, the internalization of ASC-exosomes by the cells was shown and the molecules and the mechanisms by which exosomes could exert their beneficial effect were addressed. We performed for the first time a comprehensive proteomic analysis of exosomes derived from murine ASC. We identified a total of 189 proteins and the shotgun proteomics analysis revealed that the exosomal proteins are mainly involved in cell adhesion and negative regulation of the apoptotic process. We correlated the protein content to the anti-apoptotic effect of exosomes observing a downregulation of pro-apoptotic proteins Bax and cleaved caspase-3 and upregulation of anti-apoptotic protein Bcl-2 α, in an in vitro model of ALS after cell treatment with exosomes. Overall, this study shows the neuroprotective effect of ASC-exosomes after their internalization and their global protein profile, that could be useful to understand how exosomes act, demonstrating that they can be employed as therapy in neurodegenerative diseases.

Normoxic Tumour Extracellular Vesicles Modulate the Response of Hypoxic Cancer and Stromal Cells to Doxorubicin In Vitro

“…It has been suggested for example in pancreatic cancer that TEV could promote M2 macrophage polarisation by conveying HIF-1α protein or that TEV could convey entire or cleaved Bcl-xL protein, as well as other proteins and molecules (miRNA or lncRNA) that promote the anti-apoptotic pathways after internalisation by cells, and could even lead to the production of Bcl-xL or other anti-apoptotic proteins in TEV recipient cells [ 44 , 65 , 66 ]. Thus, we further investigated whether we could detect molecular changes in the slightly less responsive hypoxic naïve C26 and RAW 264.7 cells to DOX after the pretreatment with normoxic TEV and DOX-elicited TEV ( Figure 4 C and Figure 5 C).…”

Recent Advances on Extracellular Vesicles in Central Nervous System Diseases

“…Bonafede et al reported the anti-apoptotic effect of ASC-exosomes in an ALS cell model, supported by increasing of anti-apoptotic protein and decreasing of pro-apoptotic proteins through proteomic analysis of exosomes. 96 Furthermore, neuroprotective effect of ASC-exosomes was confirmed in superoxide dismutase 1 (G93A) murine model. 97 …”

Mesenchymal stem cell-derived exosomes: Toward cell-free therapeutic strategies in regenerative medicine

“…Treatment of neurons from G93A ALS mice with human adipose-derived MSC-Exos alleviated aggregation of superoxide dismutase 1, and normalized the cellular phenotype, restoring to normal the levels of mitochondrial proteins including p-CREB and PGC-1α[ 145 ]. Subsequently, two studies by the same group showed that mouse adipose tissue-derived MSC-Exos exerted an anti-apoptotic effect and rescued the function of mitochondria in an in vitro model of ALS[ 146 , 147 ]. MS is a chronic demyelinating disease caused by CNS inflammation and immune dysfunction, which can result in severe physical disability.…”