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...
Autores principales: | , , , , , , , , , , , , , , , , , , |
---|---|
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 |
_version_ | 1784911677772267520 |
---|---|
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. |
format | Online Article Text |
id | pubmed-10036536 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT hallowswilliamc optimizinghumanagalactosidasefortreatmentoffabrydisease AT skvorakkristen optimizinghumanagalactosidasefortreatmentoffabrydisease AT agardnick optimizinghumanagalactosidasefortreatmentoffabrydisease AT krusenikki optimizinghumanagalactosidasefortreatmentoffabrydisease AT zhangxiyun optimizinghumanagalactosidasefortreatmentoffabrydisease AT zhuyu optimizinghumanagalactosidasefortreatmentoffabrydisease AT bothamrachelc optimizinghumanagalactosidasefortreatmentoffabrydisease AT chngchinping optimizinghumanagalactosidasefortreatmentoffabrydisease AT shuklacharu optimizinghumanagalactosidasefortreatmentoffabrydisease AT laojessica optimizinghumanagalactosidasefortreatmentoffabrydisease AT millermathew optimizinghumanagalactosidasefortreatmentoffabrydisease AT seroantoinette optimizinghumanagalactosidasefortreatmentoffabrydisease AT viduyajudy optimizinghumanagalactosidasefortreatmentoffabrydisease AT ismailimoulayhichamalaoui optimizinghumanagalactosidasefortreatmentoffabrydisease AT mccluskiekerryn optimizinghumanagalactosidasefortreatmentoffabrydisease AT schiffmannraphael optimizinghumanagalactosidasefortreatmentoffabrydisease AT silvermanadamp optimizinghumanagalactosidasefortreatmentoffabrydisease AT shenjinsong optimizinghumanagalactosidasefortreatmentoffabrydisease AT huismangjaltw optimizinghumanagalactosidasefortreatmentoffabrydisease |