A universal GlycoDesign for lysosomal replacement enzymes to improve circulation time and biodistribution

Yen-Hsi Chen; Weihua Tian; Makiko Yasuda; Zilu Ye; Ming Song; Ulla Mandel; Claus Kristensen; Lorenzo Povolo; André R A Marques; Tomislav Čaval; Albert J R Heck; Julio Lopes Sampaio; Ludger Johannes; Takahiro Tsukimura; Robert Desnick; Sergey Y Vakhrushev; Zhang Yang; Henrik Clausen

doi:10.3389/fbioe.2023.1128371

A universal GlycoDesign for lysosomal replacement enzymes to improve circulation time and biodistribution

Yen-Hsi Chen, Weihua Tian, Makiko Yasuda, Zilu Ye, Ming Song, Ulla Mandel, Claus Kristensen, Lorenzo Povolo, André R A Marques, Tomislav Čaval, Albert J R Heck, Julio Lopes Sampaio, Ludger Johannes, Takahiro Tsukimura, Robert Desnick, Sergey Y Vakhrushev, Zhang Yang, Henrik Clausen

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Currently available enzyme replacement therapies for lysosomal storage diseases are limited in their effectiveness due in part to short circulation times and suboptimal biodistribution of the therapeutic enzymes. We previously engineered Chinese hamster ovary (CHO) cells to produce α-galactosidase A (GLA) with various N-glycan structures and demonstrated that elimination of mannose-6-phosphate (M6P) and conversion to homogeneous sialylated N-glycans prolonged circulation time and improved biodistribution of the enzyme following a single-dose infusion into Fabry mice. Here, we confirmed these findings using repeated infusions of the glycoengineered GLA into Fabry mice and further tested whether this glycoengineering approach, Long-Acting-GlycoDesign (LAGD), could be implemented on other lysosomal enzymes. LAGD-engineered CHO cells stably expressing a panel of lysosomal enzymes [aspartylglucosamine (AGA), beta-glucuronidase (GUSB), cathepsin D (CTSD), tripeptidyl peptidase (TPP1), alpha-glucosidase (GAA) or iduronate 2-sulfatase (IDS)] successfully converted all M6P-containing N-glycans to complex sialylated N-glycans. The resulting homogenous glycodesigns enabled glycoprotein profiling by native mass spectrometry. Notably, LAGD extended the plasma half-life of all three enzymes tested (GLA, GUSB, AGA) in wildtype mice. LAGD may be widely applicable to lysosomal replacement enzymes to improve their circulatory stability and therapeutic efficacy.

Original language	English
Article number	1128371
Number of pages	18
Journal	Frontiers in Bioengineering and Biotechnology
Volume	11
DOIs	https://doi.org/10.3389/fbioe.2023.1128371
Publication status	Published - 24 Feb 2023

Bibliographical note

Publisher Copyright:
Copyright © 2023 Chen, Tian, Yasuda, Ye, Song, Mandel, Kristensen, Povolo, Marques, Čaval, Heck, Sampaio, Johannes, Tsukimura, Desnick, Vakhrushev, Yang and Clausen.

Funding

This work was supported by the Lundbeck Foundation, Novo Nordisk Foundation, Innovation Fund Denmark, and the Danish National Research Foundation (DNRF107). TČ and AH acknowledge support from the Netherlands Organization for Scientific Research (NWO) funding the Netherlands Proteomics Centre through the X-omics Road Map program (project 184.034.019) and further acknowledge the EU Horizon 2020 program INFRAIA project Epic-XS (Project 823839). LJ acknowledges support from Fondation pour la Recherche Médicale (EQU202103012926).

Funders	Funder number
Netherlands Proteomics Centre	184.034.019
Innovationsfonden
Danmarks Grundforskningsfond	DNRF107
Fondation pour la Recherche Médicale	EQU202103012926
Nederlandse Organisatie voor Wetenschappelijk Onderzoek
Lundbeckfonden
Horizon 2020	823839
Novo Nordisk Fonden

Keywords

bioengineering
enzyme replacement therapy
glycoengineering
glycoprotein therapeutics
lysosomal storage disease

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Cite this

Chen, Y.-H., Tian, W., Yasuda, M., Ye, Z., Song, M., Mandel, U., Kristensen, C., Povolo, L., Marques, A. R. A., Čaval, T., Heck, A. J. R., Sampaio, J. L., Johannes, L., Tsukimura, T., Desnick, R., Vakhrushev, S. Y., Yang, Z., & Clausen, H. (2023). A universal GlycoDesign for lysosomal replacement enzymes to improve circulation time and biodistribution. Frontiers in Bioengineering and Biotechnology, 11, Article 1128371. https://doi.org/10.3389/fbioe.2023.1128371

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abstract = "Currently available enzyme replacement therapies for lysosomal storage diseases are limited in their effectiveness due in part to short circulation times and suboptimal biodistribution of the therapeutic enzymes. We previously engineered Chinese hamster ovary (CHO) cells to produce α-galactosidase A (GLA) with various N-glycan structures and demonstrated that elimination of mannose-6-phosphate (M6P) and conversion to homogeneous sialylated N-glycans prolonged circulation time and improved biodistribution of the enzyme following a single-dose infusion into Fabry mice. Here, we confirmed these findings using repeated infusions of the glycoengineered GLA into Fabry mice and further tested whether this glycoengineering approach, Long-Acting-GlycoDesign (LAGD), could be implemented on other lysosomal enzymes. LAGD-engineered CHO cells stably expressing a panel of lysosomal enzymes [aspartylglucosamine (AGA), beta-glucuronidase (GUSB), cathepsin D (CTSD), tripeptidyl peptidase (TPP1), alpha-glucosidase (GAA) or iduronate 2-sulfatase (IDS)] successfully converted all M6P-containing N-glycans to complex sialylated N-glycans. The resulting homogenous glycodesigns enabled glycoprotein profiling by native mass spectrometry. Notably, LAGD extended the plasma half-life of all three enzymes tested (GLA, GUSB, AGA) in wildtype mice. LAGD may be widely applicable to lysosomal replacement enzymes to improve their circulatory stability and therapeutic efficacy.",

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note = "Publisher Copyright: Copyright {\textcopyright} 2023 Chen, Tian, Yasuda, Ye, Song, Mandel, Kristensen, Povolo, Marques, {\v C}aval, Heck, Sampaio, Johannes, Tsukimura, Desnick, Vakhrushev, Yang and Clausen.",

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Chen, Y-H, Tian, W, Yasuda, M, Ye, Z, Song, M, Mandel, U, Kristensen, C, Povolo, L, Marques, ARA, Čaval, T, Heck, AJR, Sampaio, JL, Johannes, L, Tsukimura, T, Desnick, R, Vakhrushev, SY, Yang, Z & Clausen, H 2023, 'A universal GlycoDesign for lysosomal replacement enzymes to improve circulation time and biodistribution', Frontiers in Bioengineering and Biotechnology, vol. 11, 1128371. https://doi.org/10.3389/fbioe.2023.1128371

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T1 - A universal GlycoDesign for lysosomal replacement enzymes to improve circulation time and biodistribution

AU - Chen, Yen-Hsi

AU - Tian, Weihua

AU - Yasuda, Makiko

AU - Ye, Zilu

AU - Song, Ming

AU - Mandel, Ulla

AU - Kristensen, Claus

AU - Povolo, Lorenzo

AU - Marques, André R A

AU - Čaval, Tomislav

AU - Heck, Albert J R

AU - Sampaio, Julio Lopes

AU - Johannes, Ludger

AU - Tsukimura, Takahiro

AU - Desnick, Robert

AU - Vakhrushev, Sergey Y

AU - Yang, Zhang

AU - Clausen, Henrik

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A universal GlycoDesign for lysosomal replacement enzymes to improve circulation time and biodistribution

Abstract

Bibliographical note

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Keywords

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