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

TY - JOUR

T1 - The glycosylation design space for recombinant lysosomal replacement enzymes produced in CHO cells

AU - Tian, Weihua

AU - Ye, Zilu

AU - Wang, Shengjun

AU - Schulz, Morten Alder

AU - Van Coillie, Julie

AU - Sun, Lingbo

AU - Chen, Yen-Hsi

AU - Narimatsu, Yoshiki

AU - Hansen, Lars

AU - Kristensen, Claus

AU - Mandel, Ulla

AU - Bennett, Eric Paul

AU - Jabbarzadeh-Tabrizi, Siamak

AU - Schiffmann, Raphael

AU - Shen, Jin-Song

AU - Vakhrushev, Sergey Y.

AU - Clausen, Henrik

AU - Yang, Zhang

PY - 2019/12/1

Y1 - 2019/12/1

N2 - 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.

AB - 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.

KW - Animals

KW - CHO Cells

KW - Cricetinae

KW - Cricetulus

KW - Disease Models, Animal

KW - Fabry Disease/drug therapy

KW - Glycosylation

KW - Lysosomes/enzymology

KW - Male

KW - Mice

KW - Mice, Knockout

KW - Recombinant Proteins/therapeutic use

KW - alpha-Galactosidase/therapeutic use

U2 - 10.1038/s41467-019-09809-3

DO - 10.1038/s41467-019-09809-3

M3 - Journal article

C2 - 31040271

SN - 2041-1723

VL - 10

JO - Nature Communications

JF - Nature Communications

M1 - 1785

ER -