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Co-opting a Bioorthogonal Reaction for Oncometabolite Detection

Zengeya, Thomas T., Garlick, Julie M., Kulkarni, Rhushikesh A., Miley, Mikayla, Roberts, Allison M., Yang, Youfeng, Crooks, Daniel R., Sourbier, Carole, Linehan, W. Marston, Meier, Jordan L.
Journal of the American Chemical Society 2016 v.138 no.49 pp. 15813-15816
alkenes, chemical reactions, chlorides, enzyme activity, fluorescence, image analysis, metabolism, metabolites, mutation, renal cell carcinoma, spectral analysis
Dysregulated metabolism is a hallmark of many diseases, including cancer. Methods to fluorescently detect metabolites have the potential to enable new approaches to cancer detection and imaging. However, fluorescent sensing methods for naturally occurring cellular metabolites are relatively unexplored. Here we report the development of a chemical approach to detect the oncometabolite fumarate. Our strategy exploits a known bioorthogonal reaction, the 1,3-dipolar cycloaddition of nitrileimines and electron-poor olefins, to detect fumarate via fluorescent pyrazoline cycloadduct formation. We demonstrate hydrazonyl chlorides serve as readily accessible nitrileimine precursors, whose reactivity and spectral properties can be tuned to enable detection of fumarate and other dipolarophile metabolites. Finally, we show this reaction can be used to detect enzyme activity changes caused by mutations in fumarate hydratase, which underlie the familial cancer predisposition syndrome hereditary leiomyomatosis and renal cell cancer. Our studies define a novel intersection of bioorthogonal chemistry and metabolite reactivity that may be harnessed to enable biological profiling, imaging, and diagnostic applications.