In vivo imaging of hydrogen peroxide production in a murine tumor model with a chemoselective bioluminescent reporter.
Living organisms produce hydrogen peroxide (H(2)O(2)) to kill invading pathogens and for cellular signaling, but aberrant generation of this reactive oxygen species is a hallmark of oxidative stress and inflammation in aging, injury, and disease. The effects of H(2)O(2) on the overall health of living animals remain elusive, in part owing to a dearth of methods for studying this transient small molecule in vivo. Here we report the design, synthesis, and in vivo applications of Peroxy Caged Luciferin-1 (PCL-1), a chemoselective bioluminescent probe for the real-time detection of H(2)O(2) within living animals. PCL-1 is a boronic acid-caged firefly luciferin molecule that selectively reacts with H(2)O(2) to release firefly luciferin, which triggers a bioluminescent response in the presence of firefly luciferase. The high sensitivity and selectivity of PCL-1 for H(2)O(2), combined with the favorable properties of bioluminescence for in vivo imaging, afford a unique technology for real-time detection of basal levels of H(2)O(2) generated in healthy, living mice. Moreover, we demonstrate the efficacy of PCL-1 for monitoring physiological fluctuations in H(2)O(2) levels by directly imaging elevations in H(2)O(2) within testosterone-stimulated tumor xenografts in vivo. The ability to chemoselectively monitor H(2)O(2) fluxes in real time in living animals offers opportunities to dissect H(2)O(2)'s disparate contributions to health, aging, and disease.
Department of Chemistry, Howard Hughes Medical Institute, University of California, Berkeley, CA 94720, USA.
Genevieve C
GC
Van de Bittner
Elena A
EA
Dubikovskaya
Carolyn R
CR
Bertozzi
Christopher J
CJ
Chang
D000818:Animals; D045744:Cell Line, Tumor; D049454:Firefly Luciferin / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D015394:Molecular Structure; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000378:metabolism* / Q000473:pathology; D014183:Transplantation, Heterologous
D005456:Fluorescent Dyes; D049454:Firefly Luciferin; D006861:Hydrogen Peroxide