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Targeted delivery of lysosomal enzymes to the endocytic compartment in human cells using engineered extracellular vesicles

Targeted delivery of lysosomal enzymes to the endocytic compartment of human cells represents a transformative technology for treating a large family of lysosomal storage diseases (LSDs). Gaucher disease is one of the most common types of LSDs caused by mutations to the lysosomal β-glucocerebrosidas...

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Autores principales: Do, Mai Anh, Levy, Daniel, Brown, Annie, Marriott, Gerard, Lu, Biao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6872767/
https://www.ncbi.nlm.nih.gov/pubmed/31754156
http://dx.doi.org/10.1038/s41598-019-53844-5
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author Do, Mai Anh
Levy, Daniel
Brown, Annie
Marriott, Gerard
Lu, Biao
author_facet Do, Mai Anh
Levy, Daniel
Brown, Annie
Marriott, Gerard
Lu, Biao
author_sort Do, Mai Anh
collection PubMed
description Targeted delivery of lysosomal enzymes to the endocytic compartment of human cells represents a transformative technology for treating a large family of lysosomal storage diseases (LSDs). Gaucher disease is one of the most common types of LSDs caused by mutations to the lysosomal β-glucocerebrosidase (GBA). Here, we describe a genetic strategy to produce engineered exosomes loaded with GBA in two different spatial configurations for targeted delivery to the endocytic compartment of recipient cells. By fusing human GBA to an exosome-anchoring protein: vesicular stomatitis virus glycoprotein (VSVG), we demonstrate that the chimeric proteins were successfully integrated into exosomes which were secreted as extracellular vesicles (EVs) by producer cells. Isolation and molecular characterization of EVs confirmed that the fusion proteins were loaded onto exosomes without altering their surface markers, particle size or distribution. Further, enzyme-loaded exosomes/EVs added to cultured medium were taken up by recipient cells. Further, the endocytosed exosomes/EVs targeted to endocytic compartments exhibited a significant increase in GBA activity. Together, we have developed a novel method for targeting and delivery of lysosomal enzymes to their natural location: the endocytic compartment of recipient cells. Since exosomes/EVs have an intrinsic ability to cross the blood-brain-barrier, our technology may provide a new approach to treat severe types of LSDs, including Gaucher disease with neurological complications.
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spelling pubmed-68727672019-12-04 Targeted delivery of lysosomal enzymes to the endocytic compartment in human cells using engineered extracellular vesicles Do, Mai Anh Levy, Daniel Brown, Annie Marriott, Gerard Lu, Biao Sci Rep Article Targeted delivery of lysosomal enzymes to the endocytic compartment of human cells represents a transformative technology for treating a large family of lysosomal storage diseases (LSDs). Gaucher disease is one of the most common types of LSDs caused by mutations to the lysosomal β-glucocerebrosidase (GBA). Here, we describe a genetic strategy to produce engineered exosomes loaded with GBA in two different spatial configurations for targeted delivery to the endocytic compartment of recipient cells. By fusing human GBA to an exosome-anchoring protein: vesicular stomatitis virus glycoprotein (VSVG), we demonstrate that the chimeric proteins were successfully integrated into exosomes which were secreted as extracellular vesicles (EVs) by producer cells. Isolation and molecular characterization of EVs confirmed that the fusion proteins were loaded onto exosomes without altering their surface markers, particle size or distribution. Further, enzyme-loaded exosomes/EVs added to cultured medium were taken up by recipient cells. Further, the endocytosed exosomes/EVs targeted to endocytic compartments exhibited a significant increase in GBA activity. Together, we have developed a novel method for targeting and delivery of lysosomal enzymes to their natural location: the endocytic compartment of recipient cells. Since exosomes/EVs have an intrinsic ability to cross the blood-brain-barrier, our technology may provide a new approach to treat severe types of LSDs, including Gaucher disease with neurological complications. Nature Publishing Group UK 2019-11-21 /pmc/articles/PMC6872767/ /pubmed/31754156 http://dx.doi.org/10.1038/s41598-019-53844-5 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Do, Mai Anh
Levy, Daniel
Brown, Annie
Marriott, Gerard
Lu, Biao
Targeted delivery of lysosomal enzymes to the endocytic compartment in human cells using engineered extracellular vesicles
title Targeted delivery of lysosomal enzymes to the endocytic compartment in human cells using engineered extracellular vesicles
title_full Targeted delivery of lysosomal enzymes to the endocytic compartment in human cells using engineered extracellular vesicles
title_fullStr Targeted delivery of lysosomal enzymes to the endocytic compartment in human cells using engineered extracellular vesicles
title_full_unstemmed Targeted delivery of lysosomal enzymes to the endocytic compartment in human cells using engineered extracellular vesicles
title_short Targeted delivery of lysosomal enzymes to the endocytic compartment in human cells using engineered extracellular vesicles
title_sort targeted delivery of lysosomal enzymes to the endocytic compartment in human cells using engineered extracellular vesicles
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6872767/
https://www.ncbi.nlm.nih.gov/pubmed/31754156
http://dx.doi.org/10.1038/s41598-019-53844-5
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