title: Co-opting a Bioorthogonal Reaction for Oncometabolite Detection.
abstract: 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.
authors.affiliation: Chemical Biology Laboratory, National Cancer Institute , Frederick, Maryland 21702, United States.
authors.forename: Thomas T
authors.initials: TT
authors.lastname: Zengeya
authors.affiliation: Chemical Biology Laboratory, National Cancer Institute , Frederick, Maryland 21702, United States.
authors.forename: Julie M
authors.initials: JM
authors.lastname: Garlick
authors.affiliation: Chemical Biology Laboratory, National Cancer Institute , Frederick, Maryland 21702, United States.
authors.forename: Rhushikesh A
authors.initials: RA
authors.lastname: Kulkarni
authors.affiliation: Chemical Biology Laboratory, National Cancer Institute , Frederick, Maryland 21702, United States.
authors.forename: Mikayla
authors.initials: M
authors.lastname: Miley
authors.affiliation: Chemical Biology Laboratory, National Cancer Institute , Frederick, Maryland 21702, United States.
authors.forename: Allison M
authors.initials: AM
authors.lastname: Roberts
authors.affiliation: Urologic Oncology Branch, National Cancer Institute , Bethesda, Maryland 20817, United States.
authors.forename: Youfeng
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Urologic Oncology Branch, National Cancer Institute , Bethesda, Maryland 20817, United States.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Crooks
authors.affiliation: Urologic Oncology Branch, National Cancer Institute , Bethesda, Maryland 20817, United States.
authors.forename: Carole
authors.initials: C
authors.lastname: Sourbier
authors.affiliation: Urologic Oncology Branch, National Cancer Institute , Bethesda, Maryland 20817, United States.
authors.forename: W Marston
authors.initials: WM
authors.lastname: Linehan
authors.affiliation: Chemical Biology Laboratory, National Cancer Institute , Frederick, Maryland 21702, United States.
authors.forename: Jordan L
authors.initials: JL
authors.lastname: Meier
mesh_terms: D000475:Alkenes / Q000737:chemistry / Q000378:metabolism*; D002292:Carcinoma, Renal Cell / Q000378:metabolism* / Q000473:pathology; D005649:Fumarate Hydratase / Q000378:metabolism*; D005650:Fumarates / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D007097:Imines / Q000737:chemistry / Q000378:metabolism*; D007680:Kidney Neoplasms / Q000378:metabolism* / Q000473:pathology; D015394:Molecular Structure
chemical_list: D000475:Alkenes; D005650:Fumarates; D007097:Imines; D005649:Fumarate Hydratase