The glycosylation design space for recombinant lysosomal replacement enzymes produced in CHO cells

Weihua Tian, Zilu Ye, Shengjun Wang, Morten Alder Schulz, Julie Van Coillie, Lingbo Sun, Yen-Hsi Chen, Yoshiki Narimatsu, Lars Hansen, Claus Kristensen, Ulla Mandel, Eric Paul Bennett, Siamak Jabbarzadeh-Tabrizi, Raphael Schiffmann, Jin-Song Shen, Sergey Y. Vakhrushev, Henrik Clausen, Zhang Yang

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

Lysosomal replacement enzymes are essential therapeutic options for rare congenital lysosomal enzyme deficiencies, but enzymes in clinical use are only partially effective due to short circulatory half-life and inefficient biodistribution. Replacement enzymes are primarily taken up by cell surface glycan receptors, and glycan structures influence uptake, biodistribution, and circulation time. It has not been possible to design and systematically study effects of different glycan features. Here we present a comprehensive gene engineering screen in Chinese hamster ovary cells that enables production of lysosomal enzymes with N-glycans custom designed to affect key glycan features guiding cellular uptake and circulation. We demonstrate distinct circulation time and organ distribution of selected glycoforms of α-galactosidase A in a Fabry disease mouse model, and find that an α2-3 sialylated glycoform designed to eliminate uptake by the mannose 6-phosphate and mannose receptors exhibits improved circulation time and targeting to hard-to-reach organs such as heart. The developed design matrix and engineered CHO cell lines enables systematic studies towards improving enzyme replacement therapeutics.

Original languageEnglish
Article number1785
JournalNature Communications
Volume10
Number of pages13
ISSN2041-1723
DOIs
Publication statusPublished - 2019

Keywords

  • Animals
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Disease Models, Animal
  • Fabry Disease/drug therapy
  • Glycosylation
  • Lysosomes/enzymology
  • Male
  • Mice
  • Mice, Knockout
  • Recombinant Proteins/therapeutic use
  • alpha-Galactosidase/therapeutic use

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