Framework engineering to produce dominant T cell receptors with enhanced antigen-specific function

Nat Commun. 2019 Oct 1;10(1):4451. doi: 10.1038/s41467-019-12441-w.

Abstract

TCR-gene-transfer is an efficient strategy to produce therapeutic T cells of defined antigen specificity. However, there are substantial variations in the cell surface expression levels of human TCRs, which can impair the function of engineered T cells. Here we demonstrate that substitutions of 3 amino acid residues in the framework of the TCR variable domains consistently increase the expression of human TCRs on the surface of engineered T cells.The modified TCRs mediate enhanced T cell proliferation, cytokine production and cytotoxicity, while reducing the peptide concentration required for triggering effector function up to 3000-fold. Adoptive transfer experiments in mice show that modified TCRs control tumor growth more efficiently than wild-type TCRs. Our data indicate that simple variable domain modifications at a distance from the antigen-binding loops lead to increased TCR expression and improved effector function. This finding provides a generic platform to optimize the efficacy of TCR gene therapy in humans.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens / immunology*
  • Antigens, CD / metabolism
  • Antigens, Differentiation, T-Lymphocyte / metabolism
  • Cell Engineering*
  • Cell Line, Tumor
  • Cell Proliferation
  • Cytokines / metabolism
  • Gene Expression
  • Genes, T-Cell Receptor / genetics*
  • Genes, T-Cell Receptor / immunology*
  • Genetic Therapy
  • Humans
  • Lectins, C-Type / metabolism
  • Lymphocyte Activation
  • Male
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Models, Molecular
  • Protein Domains
  • Protein Engineering
  • Receptors, Antigen, T-Cell / chemistry
  • Receptors, Antigen, T-Cell / genetics*
  • Receptors, Antigen, T-Cell / immunology
  • Receptors, Antigen, T-Cell / metabolism*
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism*

Substances

  • Antigens
  • Antigens, CD
  • Antigens, Differentiation, T-Lymphocyte
  • CD69 antigen
  • Cytokines
  • Lectins, C-Type
  • Receptors, Antigen, T-Cell