Fluorogenic Granzyme A Substrates Enable Real-Time Imaging of Adaptive Immune Cell Activity

Angew Chem Int Ed Engl. 2023 Feb 13;62(8):e202216142. doi: 10.1002/anie.202216142. Epub 2023 Jan 16.

Abstract

Cytotoxic immune cells, including T lymphocytes (CTLs) and natural killer (NK) cells, are essential components of the host response against tumors. CTLs and NK cells secrete granzyme A (GzmA) upon recognition of cancer cells; however, there are very few tools that can detect physiological levels of active GzmA with high spatiotemporal resolution. Herein, we report the rational design of the near-infrared fluorogenic substrates for human GzmA and mouse GzmA. These activity-based probes display very high catalytic efficiency and selectivity over other granzymes, as shown in tissue lysates from wild-type and GzmA knock-out mice. Furthermore, we demonstrate that the probes can image how adaptive immune cells respond to antigen-driven recognition of cancer cells in real time.

Keywords: Cancer; Fluorescence; Hemicyanine; Peptides; Probes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Fluorescent Dyes*
  • Granzymes
  • Humans
  • Killer Cells, Natural
  • Mice
  • Mice, Knockout
  • T-Lymphocytes, Cytotoxic*

Substances

  • Fluorescent Dyes
  • Granzymes
  • GZMA protein, human