In vivo imaging of endogenous enzyme activities using luminescent 1,2-dioxetane compounds

• Published in: Journal of biomedical science Vol. 22; no. 1; p. 45
• Main Authors: Tseng, Jen-Chieh, Kung, Andrew L
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 24.06.2015; BioMed Central
• Subjects: Alkaline Phosphatase - chemistry; Alkaline Phosphatase - metabolism; Animals; beta-Galactosidase - chemistry; beta-Galactosidase - metabolism; Diagnostic Imaging; Energy Metabolism; Enzymes; Fluorescence; Force and energy; HCT116 Cells; Heterocyclic Compounds - chemistry; Heterocyclic Compounds - pharmacology; Humans; Light; Luminescence; Mice; Phosphatases
• DOI: 10.1186/s12929-015-0155-x
• ISSN: 1423-0127; 1021-7770

Here we present a non-invasive imaging method for visualizing endogenous enzyme activities in living animals. This optical imaging method is based on an energy transfer principle termed chemically initiated electron exchange luminescence (CIEEL). The light energy is provided by enzymatic activation of metastable 1,2-dioxetane substrates, whose protective groups are removed by hydrolytic enzymes such as β-galactosidase and alkaline phosphatase. In the presence of a nearby fluorescent recipient, the chemical energy within the activated substrate is then transferred via formation of a charge-transfer complex with the fluorophore, a mechanism closely related to glow stick chemistry. Efficient CIEEL energy transfer requires close proximity between the trigger enzyme and the fluorescent recipient. Using cells stained with fluorescent dialkylcarbocyanines as the energy recipients, we demonstrated CIEEL imaging of cellular β-galactosidase or alkaline phosphatase activity. In living animals, we used a similar approach to non-invasively image alkaline phosphatase activity in the peritoneal cavity. In this report, we provide proof-of-concept for CIEEL imaging of in vivo enzymatic activity. In addition, we demonstrate the use of CIEEL energy transfer for visualizing elevated alkaline phosphatase activity associated with tissue inflammation in living animals.