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An anti-CD98 antibody displaying pH-dependent Fc-mediated tumour-specific activity against multiple cancers in CD98-humanized mice

Abstract

The cell-surface glycoprotein CD98—a subunit of the LAT1/CD98 amino acid transporter—is an attractive target for cancer immunotherapies, but its widespread expression has hampered the development of CD98-targeting antibody therapeutics. Here we report that an anti-CD98 antibody, identified via the screening of phage-display libraries of CD98 single-chain variable fragments with mutated complementarity-determining regions, preserves the physiological function of CD98 and elicits broad-spectrum crystallizable-fragment (Fc)-mediated anti-tumour activity (requiring Fcγ receptors for immunoglobulins, macrophages, dendritic cells and CD8+ T cells, as well as other components of the innate and adaptive immune systems) in multiple xenograft and syngeneic tumour models established in CD98-humanized mice. We also show that a variant of the anti-CD98 antibody with pH-dependent binding, generated by solving the structure of the antibody–CD98 complex, displayed enhanced tumour-specific activity and pharmacokinetics. pH-dependent antibody variants targeting widely expressed antigens may lead to superior therapeutic outcomes.

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Fig. 1: Anti-tumour activity of S1-F4 in human cancer-cell-line-derived xenograft tumour models.
Fig. 2: Anti-tumour activity of S1-F4 in CD98-humanized mice and C57BL/6 mice bearing syngeneic tumours.
Fig. 3: The anti-tumour activity of S1-F4 depends on Fc–FcγR interactions.
Fig. 4: Roles of different immune subsets in S1-F4’s anti-tumour efficacy.
Fig. 5: S1-F4 pharmacokinetics and biodistributions in monkeys and mice.
Fig. 6: Analyses of a S1-F4/CD98 complex crystal structure and structure-guided antibody engineering of S1-F4 to generate a pH-dependent anti-hCD98 antibody.
Fig. 7: A pH-dependent anti-hCD98 antibody displays preferential CD98 binding in tumours and has improved anti-tumour activity compared with S1-F4.

Data availability

The main data supporting the results in this study are available within the paper and its supplementary information. The structure of hCD98 ECD used in this study is available from the Protein Data Bank (PDB) under the accession code 2DH2. The S1-F4/CD98 complex crystal structure generated in this study is available from the PDB under accession code 7DF1. Source data are provided with this paper.

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Acknowledgements

We thank the NIBS Animal Facility for their help in the handling and care of mice, the NIBS Biological Resource Centre for DNA sequencing, and the NIBS imaging facility for assistance with the microscope experiment. This work was supported by grants from the Ministry of Science and Technology of the People’s Republic of China (973 Program #2012CB837600 to J.S.), the Beijing Municipal Science and Technology Commission, and the Beijing Key Laboratory of Pathogen Invasion and Immune Defense (Z171100002217064 to J.S.). The funders had no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript.

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Contributions

X.T., X.L. and J.S. conceptualized this study, interpreted the results and drafted the manuscript. X.T., X.L., K.W., J.L., Z.W., X.H., Y.L., X.W. and H.Z. performed experiments. X.L. and J.D. analysed the crystal structure. F.W. constructed the CD98-humanized mice. X.T. prepared figures. J.S. supervised the study. All authors commented on the manuscript.

Corresponding author

Correspondence to Jianhua Sui.

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Competing interests

J.S. and X.T. are co-inventors of patent applications for the antibodies reported in this study. The other authors declare no competing interests.

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Nature Biomedical Engineering thanks Kanyi Pu, John C. Zwaagstra and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary Figs. 1–10 and Tables 1–6.

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Supplementary dataset 1

Unmodified images for the gels shown in Supplementary Fig. 3b–d.

Supplementary dataset 2

Source data for the supplementary figures.

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Tian, X., Liu, X., Ding, J. et al. An anti-CD98 antibody displaying pH-dependent Fc-mediated tumour-specific activity against multiple cancers in CD98-humanized mice. Nat. Biomed. Eng (2022). https://doi.org/10.1038/s41551-022-00956-5

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