Abstract
In the present study the use of extracellular vesicles (EVs) as vehicles for therapeutic enzymes in lysosomal storage disorders was explored. EVs were isolated from mammalian cells overexpressing alpha‐galactosidase A (GLA) or N‐sulfoglucosamine sulfohydrolase (SGSH) enzymes, defective in Fabry and Sanfilippo A diseases, respectively. Direct purification of EVs from cell supernatants was found to be a simple and efficient method to obtain highly active GLA and SGSH proteins, even after EV lyophilization. Likewise, EVs carrying GLA (EV‐GLA) were rapidly uptaken and reached the lysosomes in cellular models of Fabry disease, restoring lysosomal functionality much more efficiently than the recombinant enzyme in clinical use. In vivo, EVs were well tolerated and distributed among all main organs, including the brain. DiR‐labelled EVs were localized in brain parenchyma 1 h after intra‐arterial (internal carotid artery) or intravenous (tail vein) administrations. Moreover, a single intravenous administration of EV‐GLA was able to reduce globotriaosylceramide (Gb3) substrate levels in clinically relevant tissues, such kidneys and brain. Overall, our results demonstrate that EVs from cells overexpressing lysosomal enzymes act as natural protein delivery systems, improving the activity and the efficacy of the recombinant proteins and facilitating their access to organs neglected by conventional enzyme replacement therapies.
Highlights
The use of recombinant proteins to treat a wide variety of clinical indications, including cancer, autoimmune and genetic diseases, is still a challenge (Leader et al, 2008)
In this work we aimed at testing the feasibility of extracellular vesicles (EVs) as enzyme delivery systems for galactosidase A (GLA) and sulfoglucosamine sulfohydrolase (SGSH) proteins, as a way to improve enzyme replacement therapy (ERT) in these two diseases
EVs were isolated from CHO DG44 and HEK293 cells transfected with plasmids coding for GLA and SGSH enzymes, respectively
Summary
The use of recombinant proteins to treat a wide variety of clinical indications, including cancer, autoimmune and genetic diseases, is still a challenge (Leader et al, 2008). Patient symptomatology depends on the disease and the particular mutation but often, LSDs derive in a systemic illness affecting multiple organs, including the central nervous system (CNS), liver, kidneys, heart and the musculoskeletal system. LSDs have been historically neglected by the pharmaceutical industry. Their individual low incidence has discouraged active search for treatment due to the high difficulties to organize clinical trials and poor expected post-commercialization profits. ERTs using recombinant proteins are currently available for 10 LSDs, (Concolino et al, 2018) including Fabry disease. Systemically administered enzymes are not able to reach the brain parenchyma, leaving without effective treatment LSD patients with CNS affectation (Solomon & Muro, 2017)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
