Home LiteratureArticle Details
PMID: 33063473 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

N-glycosylation of PD-1 promotes binding of camrelizumab.

EMBO reports ·Vol. 21 ·No. 12 ·2020-00-03 ·Pages e51444

Liu K, Tan S, Jin W, Guan J, Wang Q, Sun H, Qi J, Yan J, Chai Y, Wang Z, Deng C, Gao GF

Abstract

PD-1 is a highly glycosylated inhibitory receptor expressed mainly on T cells. Targeting of PD-1 with monoclonal antibodies (MAbs) to block the interaction with its ligand PD-L1 has been successful for the treatment of multiple tumors. However, polymorphisms at N-glycosylation sites of PD-1 exist in the human population that might affect antibody binding, and dysregulated glycosylation has been observed in the tumor microenvironment. Here, we demonstrate varied N-glycan composition in PD-1, and show that the binding affinity of camrelizumab, a recently approved PD-1-specific MAb, to non-glycosylated PD-1 proteins from E. coli is substantially decreased compared with glycosylated PD-1. The structure of the camrelizumab/PD-1 complex reveals that camrelizumab mainly utilizes its heavy chain to bind to PD-1, while the light chain sterically inhibits the binding of PD-L1 to PD-1. Glycosylation of asparagine 58 (N58) promotes the interaction with camrelizumab, while the efficiency of camrelizumab to inhibit the binding of PD-L1 is substantially reduced for glycosylation-deficient PD-1. These results increase our understanding of how glycosylation affects the activity of PD-1-specific MAbs during immune checkpoint therapy.

Keywords
PD-1 camrelizumab glycosylation monoclonal antibody structure
MeSH Terms
Antibodies, Monoclonal, Humanized Escherichia coli/metabolism Glycosylation Humans Programmed Cell Death 1 Receptor/genetics,metabolism
Chemicals
Antibodies, Monoclonal, Humanized Programmed Cell Death 1 Receptor camrelizumab
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Liu Kefang
Faculty of Health Sciences, University of Macau, Macau SAR, China. | CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China. | Savaid Medical School, University of Chinese Academy of Sciences, Beijing, China.
Tan Shuguang ORCID
CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Jin Wanjun
College of Life Science, Research Center for Glycobiology and Glycotechnology, College of Food Science and Technology, Northwest University, Xi'an, China.
Guan Jiawei
CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Wang Qingling
CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Sun Huan
CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Qi Jianxun
CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Yan Jinghua
CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Chai Yan
CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Wang Zhongfu
College of Life Science, Research Center for Glycobiology and Glycotechnology, College of Food Science and Technology, Northwest University, Xi'an, China.
Deng Chuxia ORCID
Faculty of Health Sciences, University of Macau, Macau SAR, China.
Gao George F ORCID
Faculty of Health Sciences, University of Macau, Macau SAR, China. | CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China. | Savaid Medical School, University of Chinese Academy of Sciences, Beijing, China.
References (42)
42 references, click to expand
  1. Adams PD, Afonine PV, Bunkoczi G, Chen VB, Davis IW, Echols N, Headd JJ, Hung LW, Kapral GJ, Grosse-Kunstleve RW et al (2010) PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr D Biol Crystallogr 66: 213-221
  2. Britain CM, Holdbrooks AT, Anderson JC, Willey CD, Bellis SL (2018) Sialylation of EGFR by the ST6Gal-I sialyltransferase promotes EGFR activation and resistance to gefitinib-mediated cell death. J Ovarian Res 11: 12
  3. Chen VB, Arendall WB III, Headd JJ, Keedy DA, Immormino RM, Kapral GJ, Murray LW, Richardson JS, Richardson DC (2010) MolProbity: all-atom structure validation for macromolecular crystallography. Acta Crystallogr D Biol Crystallogr 66: 12-21
  4. Chen D, Tan S, Zhang H, Wang H, He W, Shi R, Tong Z, Zhu J, Cheng H, Gao S et al (2019) The FG loop of PD-1 serves as a “hotspot” for therapeutic monoclonal antibodies in tumor immune checkpoint therapy. iScience 14: 113-124
  5. Collaborative Computational Project N (1994) The CCP4 suite: programs for protein crystallography. Acta Crystallogr D Biol Crystallogr 50: 760-763
  6. Emsley P, Cowtan K (2004) Coot: model-building tools for molecular graphics. Acta Crystallogr D Biol Crystallogr 60: 2126-2132
  7. Fang W, Yang Y, Ma Y, Hong S, Lin L, He X, Xiong J, Li P, Zhao H, Huang Y et al (2018) Camrelizumab (SHR-1210) alone or in combination with gemcitabine plus cisplatin for nasopharyngeal carcinoma: results from two single-arm, phase 1 trials. Lancet Oncol 19: 1338-1350
  8. Finlay WJJ, Coleman JE, Edwards JS, Johnson KS (2019) Anti-PD1 ‘SHR-1210’ aberrantly targets pro-angiogenic receptors and this polyspecificity can be ablated by paratope refinement. mAbs 11: 26-44
  9. Kleffel S, Posch C, Barthel SR, Mueller H, Schlapbach C, Guenova E, Elco CP, Lee N, Juneja VR, Zhan Q et al (2015) Melanoma cell-intrinsic PD-1 receptor functions promote tumor growth. Cell 162: 1242-1256
  10. Li L, Wang C, Qiang S, Zhao J, Song S, Jin W, Wang B, Zhang Y, Huang L, Wang Z (2016) Mass Spectrometric analysis of N-glycoforms of soybean allergenic glycoproteins separated by SDS-PAGE. J Proteome Res 64: 7367-7376
  11. Li C, Lu Y, Chen X, Yang M, Zou Z, Han J, Gao X, Tang R, Wang C, Huang L et al (2019) Analysis of the N-glycoforms and immunoactivity of Chinese yam (Dioscorea opposita Thunb.) glycoprotein 30CYGP. J Proteome Res 19: 28-35
  12. Liu WJ, Lan J, Liu K, Deng Y, Yao Y, Wu S, Chen H, Bao L, Zhang H, Zhao M et al (2017) Protective T cell responses featured by concordant recognition of middle East respiratory syndrome coronavirus-derived CD8+ T cell epitopes and host MHC. J Immunol 198: 873-882
  13. Liu H, Guo L, Zhang J, Zhou Y, Zhou J, Yao J, Wu H, Yao S, Chen B, Chai Y et al (2019) Glycosylation-independent binding of monoclonal antibody toripalimab to FG loop of PD-1 for tumor immune checkpoint therapy. mAbs 11: 681-690
  14. Liu JX, Wang GQ, Liu L, Wu RJ, Wu Y, Fang C, Zhou XH, Jiao J, Gu Y, Zhou H et al (2020) Study of the interactions of a novel monoclonal antibody, mAb059c, with the hPD-1 receptor. Sci Rep 9: 17830
  15. Lu D, Liu K, Zhang D, Yue C, Lu Q, Cheng H, Wang L, Chai Y, Qi J, Wang LF et al (2019) Peptide presentation by bat MHC class I provides new insight into the antiviral immunity of bats. PLoS Biol 17: e3000436
  16. Markham A, Keam SJ (2019) Camrelizumab: first global approval. Drugs 79: 1355-1361
  17. Munkley J, Oltean S, Vodak D, Wilson BT, Livermore KE, Zhou Y, Star E, Floros VI, Johannessen B, Knight B et al (2015) The androgen receptor controls expression of the cancer-associated sTn antigen and cell adhesion through induction of ST6GalNAc1 in prostate cancer. Oncotarget 6: 34358-34374
  18. Na Z, Yeo SP, Bharath SR, Bowler MW, Balikci E, Wang CI, Song H (2017) Structural basis for blocking PD-1-mediated immune suppression by therapeutic antibody pembrolizumab. Cell Res 27: 147-150
  19. Nanda R, Chow LQ, Dees EC, Berger R, Gupta S, Geva R, Pusztai L, Pathiraja K, Aktan G, Cheng JD et al (2016) Pembrolizumab in patients with advanced triple-negative breast cancer: phase Ib KEYNOTE-012 study. J Clin Oncol 34: 2460-2467
  20. Newsom-Davis TE, Wang D, Steinman L, Chen PF, Wang LX, Simon AK, Screaton GR (2009) Enhanced immune recognition of cryptic glycan markers in human tumors. Cancer Res 69: 2018-2025
  21. Nishino M, Ramaiya NH, Hatabu H, Hodi FS (2017) Monitoring immune-checkpoint blockade: response evaluation and biomarker development. Nat Rev Clin Oncol 14: 655-668
  22. Otwinowski Z, Minor W (1997) Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol 276: 307-326
  23. Pinho SS, Reis CA (2015) Glycosylation in cancer: mechanisms and clinical implications. Nat Rev Cancer 15: 540-555
  24. Radhakrishnan P, Dabelsteen S, Madsen FB, Francavilla C, Kopp KL, Steentoft C, Vakhrushev SY, Olsen JV, Hansen L, Bennett EP et al (2014) Immature truncated O-glycophenotype of cancer directly induces oncogenic features. Proc Natl Acad Sci USA 111: E4066-E4075
  25. Read RJ (2001) Pushing the boundaries of molecular replacement with maximum likelihood. Acta crystallogr D Biol Crystallogr 57: 1373-1382
  26. Schultz MJ, Holdbrooks AT, Chakraborty A, Grizzle WE, Landen CN, Buchsbaum DJ, Conner MG, Arend RC, Yoon KJ, Klug CA et al (2016) The tumor-associated glycosyltransferase ST6Gal-I regulates stem cell transcription factors and confers a cancer stem cell phenotype. Cancer Res 76: 3978-3988
  27. Song H, Qi J, Khedri Z, Diaz S, Yu H, Chen X, Varki A, Shi Y, Gao GF (2016) An open receptor-binding cavity of hemagglutinin-esterase-fusion glycoprotein from newly-identified Influenza D virus: basis for its broad cell tropism. PLoS Pathog 12: e1005411
  28. Song Y, Wu J, Chen X, Lin T, Cao J, Liu Y, Zhao Y, Jin J, Huang H, Hu J et al (2019) A single-arm, multicenter, phase II study of camrelizumab in relapsed or refractory classical Hodgkin lymphoma. Clin Cancer Res 25: 7363-7369
  29. Sun L, Li CW, Chung EM, Yang R, Kim YS, Park AH, Lai YJ, Yang Y, Wang YH, Liu J et al (2020) Targeting Glycosylated pd-1 induces potent antitumor immunity. Cancer Res 80: 2298-2310
  30. Tan S, Chen D, Liu K, He M, Song H, Shi Y, Liu J, Zhang CW, Qi J, Yan J et al (2016a) Crystal clear: visualizing the intervention mechanism of the PD-1/PD-L1 interaction by two cancer therapeutic monoclonal antibodies. Protein Cell 7: 866-877
  31. Tan S, Zhang CW, Gao GF (2016b) Seeing is believing: anti-PD-1/PD-L1 monoclonal antibodies in action for checkpoint blockade tumor immunotherapy. Signal Transduct Target Ther 1: 16029
  32. Tan S, Zhang H, Chai Y, Song H, Tong Z, Wang Q, Qi J, Wong G, Zhu X, Liu WJ et al (2017) An unexpected N-terminal loop in PD-1 dominates binding by nivolumab. Nat Commun 8: 14369
  33. Wang X, Chen J, Li QK, Peskoe SB, Zhang B, Choi C, Platz EA, Zhang H (2014) Overexpression of alpha (1,6) fucosyltransferase associated with aggressive prostate cancer. Glycobiology 24: 935-944
  34. Wang S, Zhang X, Wu G, Tian Z, Qian F (2017) Optimization of high-concentration endostatin formulation: harmonization of excipients’ contributions on colloidal and conformational stabilities. Inter J Pharm 530: 173-186
  35. Wang M, Wang J, Wang R, Jiao S, Wang S, Zhang S, Zhang M (2019) Identification of a monoclonal antibody that targets PD-1 in a manner requiring PD-1 Asn58 glycosylation. Commun Biol 2: 392
  36. Wang X, Yang X, Zhang C, Wang Y, Cheng T, Duan L, Tong Z, Tan S, Zhang H, Saw PE et al (2020) Tumor cell-intrinsic PD-1 receptor is a tumor suppressor and mediates resistance to PD-1 blockade therapy. Proc Natl Acad Sci USA 117: 6640-6650
  37. Xu C, Ng DT (2015) Glycosylation-directed quality control of protein folding. Nat Rev Mol Cell Biol 16: 742-752
  38. Yu J, Hubbard-Lucey VM, Tang J (2019) Immuno-oncology drug development goes global. Nat Rev Drug Discov 18: 899-900
  39. Zak KM, Kitel R, Przetocka S, Golik P, Guzik K, Musielak B, Domling A, Dubin G, Holak TA (2015) Structure of the complex of human programmed death 1, PD-1, and its ligand PD-L1. Structure 23: 2341-2348
  40. Zhang W, Qi J, Shi Y, Li Q, Gao F, Sun Y, Lu X, Lu Q, Vavricka CJ, Liu D et al (2010) Crystal structure of the swine-origin A (H1N1)-2009 influenza A virus hemagglutinin (HA) reveals similar antigenicity to that of the 1918 pandemic virus. Protein Cell 1: 459-467
  41. Zhao M, Liu K, Luo J, Tan S, Quan C, Zhang S, Chai Y, Qi J, Li Y, Bi Y et al (2018) Heterosubtypic protections against human-infecting Avian Influenza viruses correlate to biased cross-t-cell responses. mBio 9: e01408-e01418
  42. Zhu S, Liu K, Chai Y, Wu Y, Lu D, Xiao W, Cheng H, Zhao Y, Ding C, Lyu J et al (2019) Divergent peptide presentations of HLA-A(*)30 alleles revealed by structures with pathogen peptides. Front Immunol 10: 1709
Article Info
Journal
EMBO reports
Abbr.
EMBO Rep
ISSN
1469-3178
DOI
10.15252/embr.202051444
Published
2020-00-03
Epub
2020-00-15
Pages
e51444
Language
English
Region
England
NLM ID
100963049
PMCID
PMC7726772
Subset
IM
Grants
Ministry of Science and Technology of the People's Republic of China (MOST) · 2018ZX10302302-001-002
Ministry of Science and Technology of the People's Republic of China (MOST) · 2018ZX10101004-001-003
Ministry of Science and Technology of the People's Republic of China (MOST) · 2018ZX09711003-002-001
Chinese Academy of Sciences (CAS) · XDB29040201
Chair Professor Grant, University of Macau · 2019-00019-FHS
External Links
PubMed source DOI: 10.15252/embr.202051444 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2