Immune receptor inhibition through enforced phosphatase recruitment

Nature. 2020 Oct;586(7831):779-784. doi: 10.1038/s41586-020-2851-2. Epub 2020 Oct 21.

Abstract

Antibodies that antagonize extracellular receptor-ligand interactions are used as therapeutic agents for many diseases to inhibit signalling by cell-surface receptors1. However, this approach does not directly prevent intracellular signalling, such as through tonic or sustained signalling after ligand engagement. Here we present an alternative approach for attenuating cell-surface receptor signalling, termed receptor inhibition by phosphatase recruitment (RIPR). This approach compels cis-ligation of cell-surface receptors containing ITAM, ITIM or ITSM tyrosine phosphorylation motifs to the promiscuous cell-surface phosphatase CD452,3, which results in the direct intracellular dephosphorylation of tyrosine residues on the receptor target. As an example, we found that tonic signalling by the programmed cell death-1 receptor (PD-1) results in residual suppression of T cell activation, but is not inhibited by ligand-antagonist antibodies. We engineered a PD-1 molecule, which we denote RIPR-PD1, that induces cross-linking of PD-1 to CD45 and inhibits both tonic and ligand-activated signalling. RIPR-PD1 demonstrated enhanced inhibition of checkpoint blockade compared with ligand blocking by anti-PD1 antibodies, and increased therapeutic efficacy over anti-PD1 in mouse tumour models. We also show that the RIPR strategy extends to other immune-receptor targets that contain activating or inhibitory ITIM, ITSM or ITAM motifs; for example, inhibition of the macrophage SIRPα 'don't eat me' signal with a SIRPα-CD45 RIPR molecule potentiates antibody-dependent cellular phagocytosis beyond that of SIRPα blockade alone. RIPR represents a general strategy for direct attenuation of signalling by kinase-activated cell-surface receptors.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal, Humanized / pharmacology
  • Carcinoma, Small Cell / drug therapy
  • Carcinoma, Small Cell / metabolism
  • Carcinoma, Small Cell / pathology
  • Cell Line, Tumor
  • Colonic Neoplasms / drug therapy
  • Colonic Neoplasms / metabolism
  • Colonic Neoplasms / pathology
  • Cross-Linking Reagents
  • Disease Models, Animal
  • Disease Progression
  • Female
  • HEK293 Cells
  • Humans
  • Leukocyte Common Antigens / antagonists & inhibitors
  • Leukocyte Common Antigens / chemistry
  • Leukocyte Common Antigens / metabolism*
  • Ligands
  • Lymphocyte Activation / drug effects
  • Male
  • Mice
  • Nivolumab / pharmacology
  • Phosphoric Monoester Hydrolases / metabolism*
  • Phosphorylation
  • Programmed Cell Death 1 Receptor / antagonists & inhibitors
  • Receptors, Immunologic / antagonists & inhibitors*
  • Signal Transduction / drug effects
  • T-Lymphocytes / cytology
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / immunology

Substances

  • Antibodies, Monoclonal, Humanized
  • Cross-Linking Reagents
  • Ligands
  • PDCD1 protein, human
  • Programmed Cell Death 1 Receptor
  • Receptors, Immunologic
  • Nivolumab
  • pembrolizumab
  • Phosphoric Monoester Hydrolases
  • Leukocyte Common Antigens
  • PTPRC protein, human