Activity-based bioluminescence probes for in vivo sensing applications

Abstract

Bioluminescence imaging is a highly sensitive technique commonly used for various in vivo applications. Recent efforts to expand the utility of this modality have led to the development of a suite of activity-based sensing (ABS) probes for bioluminescence imaging by 'caging' of luciferin and its structural analogs. The ability to selectively detect a given biomarker has presented researchers with many exciting opportunities to study both health and disease states in animal models. Here, we highlight recent (2021-2023) bioluminescence-based ABS probes with an emphasis on probe design and in vivo validation experiments.

Figure 1.

a) Schematic representing the sequential reaction between GBLI-1 or GBLI-2 with granzyme-B and luciferase to produce bioluminescence. b) The ratios of GzmB to Dluc signals upon rechallenging cured mice with CT26-luc cells (top) or 4T1-luc cells (bottom). c) Schematic showing the release of aminoluciferin upon sequential activation of BL-FAP by FAP and luciferase. d) Representative images of mice treated with a vehicle control, BL-FAP, or BL-FAP + inhibitor. Figure 1b was reprinted (adapted) with permission from Cell Chemical Biology 2022, 29, 1556–1567. Copyright (2022) Elsevier. Figure 1d was reprinted (adapted) with permission from Analytical Biochemistry 2022, 655, 114859. Copyright (2022) Elsevier.

Figure 2.

a) Schematic illustrating the reaction between NBP with NE to afford luciferin, which further reacts with luciferase to produce bioluminescence. b) Representative bioluminescence images of transgenic mice pre-treated with a vehicle control or fluoxetine (100 mg/mL). ROI represents brain region. c) Quantification of bioluminescence from transgenic mice treated with a vehicle control or NE. d) Schematic illustrating the reaction between P Probe and superoxide anion. e) In vitro assay demonstrating 18.5-fold signal enhancement upon treatment of P Probe with O₂^.−. f) Representative images of mice treated with PBS and cisplatin at 2 or 4 mg/mL, prior to P Probe administration. Figure 2b and 2c were reprinted (adapted) with permission from Anal. Chem. 2022, 94, 6441−6445. Copyright (2022) American Chemical Society. Figure 2e and 2f were reprinted (adapted) with permission from Biosensors and Bioelectronics 2022, 216, 114632. Copyright (2022) Elsevier.

Figure 3.

a) Schematic illustrating NO-mediated activation of BL660-NO. b) Experimental design showing mice on high-fat or low-fat diets being implanted with 4T1-luc followed by administration of BL660-NO. c) Quantified data of bioluminescence signals from the 24-week diet study. d) Schematic illustrating activation of BL660-NTR via nitroreductase activity. e) Representative images of mice showing activation of the trigger compared to signal due to background hydrolysis. Figures 3b and 3c were reprinted (adapted) with permission from ACS Cent. Sci. 2022, 8, 461−472. Copyright (2022) American Chemical Society. Figures 3d and 3e were reprinted (adapted) with permission from J. Am. Chem. Soc. 2023, 145, 1460−1469. Copyright (2023) American Chemical Society.

Figure 4.

a) Cartoon representation of mice harboring mutant bacteria containing Fluc-LRE genes. b) Schematic representing reactions of luciferin with Fluc and regeneration of the substrate in presence of LRE and D-cysteine. c) Representative bioluminescence images of mice harboring the mutant bacteria. d) Structure of masked bioluminescence substrate ETZ. e) Structural representation of mutant enzyme BRIC. f) Representative images of live mice with the hippocampus transduced with BRIC or OCaMBI110 AAVs. Figures 4a, 4b and 4c were reprinted (adapted) with permission from Anal. Chem. 2021, 93, 15687−15695. Copyright (2021) American Chemical Society. Figures 4e and 4f were reprinted (adapted) with permission from Nat. Commun. 2022, 13, 3967. Copyright (2022) Springer Nature.