Removal of N-Linked Glycosylation Enhances PD-L1 Detection and Predicts Anti-PD-1/PD-L1 Therapeutic Efficacy

• Published in: Cancer cell Vol. 36; no. 2; pp. 168 - 178.e4
• Main Authors: Lee, Heng-Huan, Wang, Ying-Nai, Xia, Weiya, Chen, Chia-Hung, Rau, Kun-Ming, Ye, Leiguang, Wei, Yongkun, Chou, Chao-Kai, Wang, Shao-Chun, Yan, Meisi, Tu, Chih-Yen, Hsia, Te-Chun, Chiang, Shu-Fen, Chao, K.S. Clifford, Wistuba, Ignacio I., Hsu, Jennifer L., Hortobagyi, Gabriel N., Hung, Mien-Chie
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 12.08.2019
• Subjects: A549 Cells; Antibodies - immunology; Antibody Specificity; antibody-based detection; Antineoplastic Agents, Immunological - therapeutic use; B7-H1 Antigen - antagonists & inhibitors; B7-H1 Antigen - immunology; B7-H1 Antigen - metabolism; biomarker; Clinical Decision-Making; False Negative Reactions; Glycosylation; glycosylation/deglycosylation; heterogeneity/homogeneity; Humans; immune checkpoint; Immunohistochemistry; immunotherapy; Jurkat Cells; MCF-7 Cells; Neoplasms - drug therapy; Neoplasms - immunology; Neoplasms - metabolism; Patient Selection; PD-1; PD-L1; Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase - metabolism; Predictive Value of Tests; Protein Processing, Post-Translational; Reproducibility of Results; Specimen Handling - methods; THP-1 Cells; glycosylation/deglycosylation; immune checkpoint; immunotherapy; PD-L1; immunohistochemistry; biomarker; PD-1; antibody-based detection; heterogeneity/homogeneity
• ISSN: 1535-6108; 1878-3686; 1878-3686
• DOI: 10.1016/j.ccell.2019.06.008

Reactivation of T cell immunity by PD-1/PD-L1 immune checkpoint blockade has been shown to be a promising cancer therapeutic strategy. However, PD-L1 immunohistochemical readout is inconsistent with patient response, which presents a clinical challenge to stratify patients. Because PD-L1 is heavily glycosylated, we developed a method to resolve this by removing the glycan moieties from cell surface antigens via enzymatic digestion, a process termed sample deglycosylation. Notably, deglycosylation significantly improves anti-PD-L1 antibody binding affinity and signal intensity, resulting in more accurate PD-L1 quantification and prediction of clinical outcome. This proposed method of PD-L1 antigen retrieval may provide a practical and timely approach to reduce false-negative patient stratification for guiding anti-PD-1/PD-L1 therapy. [Display omitted] •N-linked glycosylation of PD-L1 hinders its recognition by PD-L1 antibodies•Removal of glycosylation enhances anti-PD-L1 signal in a variety of bioassays•Patient sample deglycosylation prevents false-negative detection of PD-L1•Deglycosylated PD-L1 is a more reliable biomarker to guide immunotherapy Histological detection of PD-L1 may guide therapy with anti-PD-1/PD-L1 antibodies but some PD-L1-negative tumors respond to these treatments. Lee et al. show that enzymatic deglycosylation of tissue sections improves PD-L1 detection and its predictive value, and could potentially impact patient stratification.