Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

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

Nature Biomedical Engineering volume 7, pages 8–23 (2023)Cite this article

Subjects

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.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

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.

Similar content being viewed by others

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.

References

  1. Mastroberardino, L. et al. Amino-acid transport by heterodimers of 4F2hc/CD98 and members of a permease family. Nature 395, 288–291 (1998).

    Article  CAS  Google Scholar 

  2. Nakamura, E. et al. 4F2 (CD98) heavy chain is associated covalently with an amino acid transporter and controls intracellular trafficking and membrane topology of 4F2 heterodimer. J. Biol. Chem. 274, 3009–3016 (1999).

    Article  CAS  Google Scholar 

  3. Rosell, A. et al. Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hc. Proc. Natl Acad. Sci. USA 111, 2966–2971 (2014).

    Article  CAS  Google Scholar 

  4. Yan, R., Zhao, X., Lei, J. & Zhou, Q. Structure of the human LAT1–4F2hc heteromeric amino acid transporter complex. Nature 568, 126–130 (2019).

    Article  Google Scholar 

  5. Nicklin, P. et al. Bidirectional transport of amino acids regulates mTOR and autophagy. Cell 136, 521–534 (2009).

    Article  CAS  Google Scholar 

  6. Cormerais, Y. et al. Genetic disruption of the multifunctional CD98/LAT1 complex demonstrates the key role of essential amino acid transport in the control of mTORC1 and tumor growth. Cancer Res. 76, 4481–4492 (2016).

    Article  CAS  Google Scholar 

  7. Timmerman, L. A. et al. Glutamine sensitivity analysis identifies the xCT antiporter as a common triple-negative breast tumor therapeutic target. Cancer Cell 24, 450–465 (2013).

    Article  CAS  Google Scholar 

  8. Harris, I. S. et al. Glutathione and thioredoxin antioxidant pathways synergize to drive cancer initiation and progression. Cancer Cell 27, 211–222 (2015).

    Article  CAS  Google Scholar 

  9. Bajaj, J. et al. CD98-mediated adhesive signaling enables the establishment and propagation of acute myelogenous leukemia. Cancer Cell 30, 792–805 (2016).

    Article  CAS  Google Scholar 

  10. Canup, B. S. B., Song, H. & Laroui, H. Role of CD98 in liver disease. Ann. Hepatol. 19, 602–607 (2020).

    Article  CAS  Google Scholar 

  11. Kaira, K. et al. CD98 expression is associated with poor prognosis in resected non-small-cell lung cancer with lymph node metastases. Ann. Surg. Oncol. 16, 3473–3481 (2009).

    Article  Google Scholar 

  12. Furuya, M., Horiguchi, J., Nakajima, H., Kanai, Y. & Oyama, T. Correlation of L-type amino acid transporter 1 and CD98 expression with triple negative breast cancer prognosis. Cancer Sci. 103, 382–389 (2012).

    Article  CAS  Google Scholar 

  13. Toyoda, M. et al. Prognostic significance of amino-acid transporter expression (LAT1, ASCT2, and xCT) in surgically resected tongue cancer. Br. J. Cancer 110, 2506–2513 (2014).

    Article  CAS  Google Scholar 

  14. Toyoda, M. et al. CD98 as a novel prognostic indicator for patients with stage III/IV hypopharyngeal squamous cell carcinoma. Head Neck 37, 1569–1574 (2015).

    Article  Google Scholar 

  15. Theodosakis, N. et al. Integrative discovery of CD98 as a melanoma biomarker. Pigm. Cell Melanoma Res. 29, 385–387 (2016).

    Article  Google Scholar 

  16. Kaira, K. et al. Prognostic significance of L-type amino-acid transporter 1 expression in surgically resected pancreatic cancer. Br. J. Cancer 107, 632–638 (2012).

    Article  CAS  Google Scholar 

  17. Xiao, B. et al. Silencing of intestinal glycoprotein CD98 by orally targeted nanoparticles enhances chemosensitization of colon cancer. ACS Nano 12, 5253–5265 (2018).

    Article  CAS  Google Scholar 

  18. Arndt, C. et al. UniCAR T cell immunotherapy enables efficient elimination of radioresistant cancer cells. Oncoimmunology 9, 1743036 (2020).

    Article  Google Scholar 

  19. Hayes, G. M. et al. Antitumor activity of an anti-CD98 antibody. Int. J. Cancer 137, 710–720 (2015).

    Article  CAS  Google Scholar 

  20. Rossier, G. et al. LAT2, a new basolateral 4F2hc/CD98-associated amino acid transporter of kidney and intestine. J. Biol. Chem. 274, 34948–34954 (1999).

    Article  CAS  Google Scholar 

  21. Fagerberg, L. et al. Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibody-based proteomics. Mol. Cell. Proteom. 13, 397–406 (2014).

    Article  CAS  Google Scholar 

  22. Cantor, J. et al. CD98hc facilitates B cell proliferation and adaptive humoral immunity. Nat. Immunol. 10, 412–419 (2009).

    Article  CAS  Google Scholar 

  23. Tsumura, H. et al. The targeted disruption of the CD98 gene results in embryonic lethality. Biochem. Biophys. Res. Commun. 308, 847–851 (2003).

    Article  CAS  Google Scholar 

  24. Li, D. et al. A potent human neutralizing antibody Fc-dependently reduces established HBV infections. eLife 6, e26738 (2017).

    Article  Google Scholar 

  25. Chao, M. P. et al. Anti-CD47 antibody synergizes with rituximab to promote phagocytosis and eradicate non-Hodgkin lymphoma. Cell 142, 699–713 (2010).

    Article  CAS  Google Scholar 

  26. Grillo-Lopez, A. J. et al. Rituximab: the first monoclonal antibody approved for the treatment of lymphoma. Curr. Pharm. Biotechnol. 1, 1–9 (2000).

    Article  CAS  Google Scholar 

  27. Scott, A. M., Wolchok, J. D. & Old, L. J. Antibody therapy of cancer. Nat. Rev. Cancer 12, 278–287 (2012).

    Article  CAS  Google Scholar 

  28. Wilson, N. S. et al. An Fcgamma receptor-dependent mechanism drives antibody-mediated target-receptor signaling in cancer cells. Cancer cell 19, 101–113 (2011).

    Article  CAS  Google Scholar 

  29. Shields, R. L. et al. High resolution mapping of the binding site on human IgG1 for Fc gamma RI, Fc gamma RII, Fc gamma RIII, and FcRn and design of IgG1 variants with improved binding to the Fc gamma R. J. Biol. Chem. 276, 6591–6604 (2001).

    Article  CAS  Google Scholar 

  30. Idusogie, E. E. et al. Mapping of the C1q binding site on rituxan, a chimeric antibody with a human IgG1 Fc. J. Immunol. 164, 4178–4184 (2000).

    Article  CAS  Google Scholar 

  31. Treffers, L. W. et al. FcgammaRIIIb restricts antibody-dependent destruction of cancer cells by human neutrophils. Front. Immunol. 9, 3124 (2018).

    Article  CAS  Google Scholar 

  32. Nimmerjahn, F., Gordan, S. & Lux, A. FcgammaR dependent mechanisms of cytotoxic, agonistic, and neutralizing antibody activities. Trends Immunol. 36, 325–336 (2015).

    Article  CAS  Google Scholar 

  33. Seidel, U. J., Schlegel, P. & Lang, P. Natural killer cell mediated antibody-dependent cellular cytotoxicity in tumor immunotherapy with therapeutic antibodies. Front. Immunol. 4, 76 (2013).

    Article  Google Scholar 

  34. Kamber, R. A. et al. Inter-cellular CRISPR screens reveal regulators of cancer cell phagocytosis. Nature 597, 549–554 (2021).

    Article  CAS  Google Scholar 

  35. Yang, X. et al. Cetuximab-mediated tumor regression depends on innate and adaptive immune responses. Mol. Ther. 21, 91–100 (2013).

    Article  CAS  Google Scholar 

  36. Liu, X. et al. CD47 blockade triggers T cell-mediated destruction of immunogenic tumors. Nat. Med. 21, 1209–1215 (2015).

    Article  CAS  Google Scholar 

  37. Vander Heiden, M. G., Cantley, L. C. & Thompson, C. B. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324, 1029–1033 (2009).

    Article  Google Scholar 

  38. Warburg, O. On the origin of cancer cells. Science 123, 309–314 (1956).

    Article  CAS  Google Scholar 

  39. Corbet, C. & Feron, O. Tumour acidosis: from the passenger to the driver’s seat. Nat. Rev. Cancer 17, 577–593 (2017).

    Article  CAS  Google Scholar 

  40. Gatenby, R. A. & Gillies, R. J. Why do cancers have high aerobic glycolysis? Nat. Rev. Cancer 4, 891–899 (2004).

    Article  CAS  Google Scholar 

  41. Robey, I. F. et al. Bicarbonate increases tumor pH and inhibits spontaneous metastases. Cancer Res. 69, 2260–2268 (2009).

    Article  CAS  Google Scholar 

  42. Helmlinger, G., Yuan, F., Dellian, M. & Jain, R. K. Interstitial pH and pO2 gradients in solid tumors in vivo: high-resolution measurements reveal a lack of correlation. Nat. Med. 3, 177–182 (1997).

    Article  CAS  Google Scholar 

  43. Sarkar, C. A. et al. Rational cytokine design for increased lifetime and enhanced potency using pH-activated ‘histidine switching’. Nat. Biotechnol. 20, 908–913 (2002).

    Article  CAS  Google Scholar 

  44. Johnston, R. J. et al. VISTA is an acidic pH-selective ligand for PSGL-1. Nature 574, 565–570 (2019).

    Article  CAS  Google Scholar 

  45. Chaparro-Riggers, J. et al. Increasing serum half-life and extending cholesterol lowering in vivo by engineering antibody with pH-sensitive binding to PCSK9. J. Biol. Chem. 287, 11090–11097 (2012).

    Article  CAS  Google Scholar 

  46. Ramanathan, S. & Jagannathan, N. Tumor associated macrophage: a review on the phenotypes, traits and functions. Iran. J. Cancer Prev. 7, 1–8 (2014).

    CAS  Google Scholar 

  47. Feral, C. C. et al. CD98hc (SLC3A2) mediates integrin signaling. Proc. Natl Acad. Sci. USA 102, 355–360 (2005).

    Article  CAS  Google Scholar 

  48. Lee, Y. et al. Cryo-EM structure of the human L-type amino acid transporter 1 in complex with glycoprotein CD98hc. Nat. Struct. Mol. Biol. 26, 510–517 (2019).

    Article  CAS  Google Scholar 

  49. Igawa, T. et al. Antibody recycling by engineered pH-dependent antigen binding improves the duration of antigen neutralization. Nat. Biotechnol. 28, 1203–1207 (2010).

    Article  CAS  Google Scholar 

  50. Kang, J. C. et al. Engineering a HER2-specific antibody–drug conjugate to increase lysosomal delivery and therapeutic efficacy. Nat. Biotechnol. 37, 523–526 (2019).

    Article  CAS  Google Scholar 

  51. Sulea, T. et al. Structure-based engineering of pH-dependent antibody binding for selective targeting of solid-tumor microenvironment. MAbs 12, 1682866 (2020).

    Article  Google Scholar 

  52. Zuchero, Y. J. et al. Discovery of novel blood–brain barrier targets to enhance brain uptake of therapeutic antibodies. Neuron 89, 70–82 (2016).

    Article  CAS  Google Scholar 

  53. Ishiguro, T. et al. Anti-glypican 3 antibody as a potential antitumor agent for human liver cancer. Cancer Res. 68, 9832–9838 (2008).

    Article  CAS  Google Scholar 

  54. Wang, Q. S. et al. The macromolecular crystallography beamline of SSRF. Nucl. Sci. Tech. 26, 12–17 (2015).

    Google Scholar 

  55. Fort, J. et al. The structure of human 4F2hc ectodomain provides a model for homodimerization and electrostatic interaction with plasma membrane. J. Biol. Chem. 282, 31444–31452 (2007).

    Article  CAS  Google Scholar 

Download references

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.

Author information

Author notes

  1. These authors contributed equally: Xinxin Tian, Ximing Liu.

Authors and Affiliations

  1. National Institute of Biological Sciences, Beijing, China

    Xinxin Tian, Ximing Liu, Jingjin Ding, Fengchao Wang, Kailun Wang, Juan Liu, Zhizhong Wei, Xinyan Hao, Yulu Li, Xin Wei, Huixiang Zhang & Jianhua Sui

  2. Peking University–Tsinghua University–National Institute of Biological Sciences (PTN) Joint Graduate Program, Peking University, Beijing, China

    Xinxin Tian & Ximing Liu

  3. National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China

    Jingjin Ding

  4. Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua University, Beijing, China

    Jianhua Sui

Authors
  1. Xinxin Tian
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ximing Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jingjin Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Fengchao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kailun Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Juan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Zhizhong Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xinyan Hao
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Yulu Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Xin Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Huixiang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Jianhua Sui
    View author publications

    You can also search for this author in PubMed Google Scholar

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.

Ethics declarations

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.

Peer review

Peer review information

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.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

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

Reporting Summary

Supplementary dataset 1

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

Supplementary dataset 2

Source data for the supplementary figures.

Source data

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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 7, 8–23 (2023). https://doi.org/10.1038/s41551-022-00956-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41551-022-00956-5

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing