Abstract
Natural killer (NK) cells are immune cells that can kill certain types of cancer cells. Adoptive transfer of NK cells represents a promising immunotherapy for malignant tumours; however, there is a lack of methods to validate anti‐tumour activity of NK cells in vivo. Herein, we report a new chemiluminescent probe to image in situ the granzyme B‐mediated killing activity of NK cells against cancer cells. We have optimised a granzyme B‐specific construct using an activatable phenoxydioxetane reporter so that enzymatic cleavage of the probe results in bright chemiluminescence. The probe shows high selectivity for active granzyme B over other proteases and higher signal‐to‐noise ratios than commercial fluorophores. Finally, we demonstrate that the probe can detect NK cell activity in mouse models, being the first chemiluminescent probe for in vivo imaging of NK cell activity in live tumours.
Highlights
Upon recognition of target tumour cells, Natural killer (NK) cells release cytolytic granzymes into target cells, where they induce apoptosis.[13–15] The tumour-killing capacity of NK cells can be monitored in vitro by different methods
We developed the peptide-based chemiluminescence probe 1 utilising the sequence Ile-Glu-Pro-Asp (IEPD), which was reported by Thornberry et al as a substrate for granzyme B.[35]
Probe 1 is composed of 3 chemical moieties: 1) the granzyme B-cleavable peptide sequence AcIEPD, 2) a self-immolative p-aminobenzyl alcohol (PABA) spacer including an electrophilic amide bond to favour enzymatic reactivity, and 3) a phenoxydioxetane moiety that emits chemiluminescence upon cleavage of the construct
Summary
A Functional Chemiluminescent Probe for in vivo Imaging of Natural Killer Cell Activity against Tumours. Citation for published version: Scott, J, Gutkin, S, Green, O, Thompson, EJ, Kitamura, T, Shabat, D & Vendrell, M 2020, 'A Functional Chemiluminescent Probe for in vivo Imaging of Natural Killer Cell Activity against Tumours', Angewandte Chemie International Edition.
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