Optimizing human α-galactosidase for treatment of Fabry disease

Fabry disease is caused by a deficiency of α-galactosidase A (GLA) leading to the lysosomal accumulation of globotriaosylceramide (Gb3) and other glycosphingolipids. Fabry patients experience significant damage to the heart, kidney, and blood vessels that can be fatal. Here we apply directed evoluti...

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Autores principales: Hallows, William C., Skvorak, Kristen, Agard, Nick, Kruse, Nikki, Zhang, Xiyun, Zhu, Yu, Botham, Rachel C., Chng, Chinping, Shukla, Charu, Lao, Jessica, Miller, Mathew, Sero, Antoinette, Viduya, Judy, Ismaili, Moulay Hicham Alaoui, McCluskie, Kerryn, Schiffmann, Raphael, Silverman, Adam P., Shen, Jin-Song, Huisman, Gjalt W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10036536/
https://www.ncbi.nlm.nih.gov/pubmed/36959353
http://dx.doi.org/10.1038/s41598-023-31777-4
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author Hallows, William C.
Skvorak, Kristen
Agard, Nick
Kruse, Nikki
Zhang, Xiyun
Zhu, Yu
Botham, Rachel C.
Chng, Chinping
Shukla, Charu
Lao, Jessica
Miller, Mathew
Sero, Antoinette
Viduya, Judy
Ismaili, Moulay Hicham Alaoui
McCluskie, Kerryn
Schiffmann, Raphael
Silverman, Adam P.
Shen, Jin-Song
Huisman, Gjalt W.
author_facet Hallows, William C.
Skvorak, Kristen
Agard, Nick
Kruse, Nikki
Zhang, Xiyun
Zhu, Yu
Botham, Rachel C.
Chng, Chinping
Shukla, Charu
Lao, Jessica
Miller, Mathew
Sero, Antoinette
Viduya, Judy
Ismaili, Moulay Hicham Alaoui
McCluskie, Kerryn
Schiffmann, Raphael
Silverman, Adam P.
Shen, Jin-Song
Huisman, Gjalt W.
author_sort Hallows, William C.
collection PubMed
description Fabry disease is caused by a deficiency of α-galactosidase A (GLA) leading to the lysosomal accumulation of globotriaosylceramide (Gb3) and other glycosphingolipids. Fabry patients experience significant damage to the heart, kidney, and blood vessels that can be fatal. Here we apply directed evolution to generate more stable GLA variants as potential next generation treatments for Fabry disease. GLAv05 and GLAv09 were identified after screening more than 12,000 GLA variants through 8 rounds of directed evolution. Both GLAv05 and GLAv09 exhibit increased stability at both lysosomal and blood pH, stability to serum, and elevated enzyme activity in treated Fabry fibroblasts (19-fold) and GLA(–/–) podocytes (10-fold). GLAv05 and GLAv09 show improved pharmacokinetics in mouse and non-human primates. In a Fabry mouse model, the optimized variants showed prolonged half-lives in serum and relevant tissues, and a decrease of accumulated Gb3 in heart and kidney. To explore the possibility of diminishing the immunogenic potential of rhGLA, amino acid residues in sequences predicted to bind MHC II were targeted in late rounds of GLAv09 directed evolution. An MHC II-associated peptide proteomics assay confirmed a reduction in displayed peptides for GLAv09. Collectively, our findings highlight the promise of using directed evolution to generate enzyme variants for more effective treatment of lysosomal storage diseases.
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spelling pubmed-100365362023-03-25 Optimizing human α-galactosidase for treatment of Fabry disease Hallows, William C. Skvorak, Kristen Agard, Nick Kruse, Nikki Zhang, Xiyun Zhu, Yu Botham, Rachel C. Chng, Chinping Shukla, Charu Lao, Jessica Miller, Mathew Sero, Antoinette Viduya, Judy Ismaili, Moulay Hicham Alaoui McCluskie, Kerryn Schiffmann, Raphael Silverman, Adam P. Shen, Jin-Song Huisman, Gjalt W. Sci Rep Article Fabry disease is caused by a deficiency of α-galactosidase A (GLA) leading to the lysosomal accumulation of globotriaosylceramide (Gb3) and other glycosphingolipids. Fabry patients experience significant damage to the heart, kidney, and blood vessels that can be fatal. Here we apply directed evolution to generate more stable GLA variants as potential next generation treatments for Fabry disease. GLAv05 and GLAv09 were identified after screening more than 12,000 GLA variants through 8 rounds of directed evolution. Both GLAv05 and GLAv09 exhibit increased stability at both lysosomal and blood pH, stability to serum, and elevated enzyme activity in treated Fabry fibroblasts (19-fold) and GLA(–/–) podocytes (10-fold). GLAv05 and GLAv09 show improved pharmacokinetics in mouse and non-human primates. In a Fabry mouse model, the optimized variants showed prolonged half-lives in serum and relevant tissues, and a decrease of accumulated Gb3 in heart and kidney. To explore the possibility of diminishing the immunogenic potential of rhGLA, amino acid residues in sequences predicted to bind MHC II were targeted in late rounds of GLAv09 directed evolution. An MHC II-associated peptide proteomics assay confirmed a reduction in displayed peptides for GLAv09. Collectively, our findings highlight the promise of using directed evolution to generate enzyme variants for more effective treatment of lysosomal storage diseases. Nature Publishing Group UK 2023-03-23 /pmc/articles/PMC10036536/ /pubmed/36959353 http://dx.doi.org/10.1038/s41598-023-31777-4 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Hallows, William C.
Skvorak, Kristen
Agard, Nick
Kruse, Nikki
Zhang, Xiyun
Zhu, Yu
Botham, Rachel C.
Chng, Chinping
Shukla, Charu
Lao, Jessica
Miller, Mathew
Sero, Antoinette
Viduya, Judy
Ismaili, Moulay Hicham Alaoui
McCluskie, Kerryn
Schiffmann, Raphael
Silverman, Adam P.
Shen, Jin-Song
Huisman, Gjalt W.
Optimizing human α-galactosidase for treatment of Fabry disease
title Optimizing human α-galactosidase for treatment of Fabry disease
title_full Optimizing human α-galactosidase for treatment of Fabry disease
title_fullStr Optimizing human α-galactosidase for treatment of Fabry disease
title_full_unstemmed Optimizing human α-galactosidase for treatment of Fabry disease
title_short Optimizing human α-galactosidase for treatment of Fabry disease
title_sort optimizing human α-galactosidase for treatment of fabry disease
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10036536/
https://www.ncbi.nlm.nih.gov/pubmed/36959353
http://dx.doi.org/10.1038/s41598-023-31777-4
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