CAREER: Diazaborine-Based Tools and Platforms for Mapping Intra- and Extracellular Peroxynitrite

With the support of the Chemistry of Life Processes program in the Division of Chemistry, Dylan Domaille from the Colorado School of Mines is studying the development of new fluorescent compounds and materials to track peroxynitrite in cellular systems. Peroxynitrite is a potent oxidant produced by live cells that can damage biomolecules like DNA and proteins. However, peroxynitrite is challenging to track in complex biological systems without interference from other compounds because of its high reactivity and short lifetime. Dr. Domaille will develop a new chemical ‘switch’ that reacts selectively with peroxynitrite. This chemical switch will activate the fluorescence of another compound, yielding a new class of fluorescent probes that illuminate where and when peroxynitrite is produced by cells. In pursuit of these goals, undergraduate and graduate students will learn synthetic chemistry, fluorescence spectroscopy, and live-cell fluorescence microscopy. In a complementary, two-pronged, educational program focused on scientific communication, undergraduate students will learn how to present information and ideas varying from scientific data to biotechnological pitches to a broad range of audiences, including professional scientists, student peers, and high school students. This research project will pursue the synthesis and evaluation of diazaborine-based intracellular fluorescent probes for tracking peroxynitrite with resolution that makes possible measurement in different cell compartments. New imaging platforms will also be developed to track extracellular peroxynitrite with spatial fidelity in pursuit of single-cell extracellular metabolite imaging. These tools will be used to understand how peroxynitrite profiles change in response to mechanical cues (e.g., stiffness, viscoelasticity, etc.) that are present in living tissue but are not captured in traditional cell culture. The proposed work aims to a) establish diazaborines as peroxynitrite-specific motifs; b) invent new technologies for imaging intra- and extracellular peroxynitrite; and c) understand how mechanical cues impact intra- and extracellular oxidative metabolite profiles. Results from this program are expected to provide a foundation for the development of peroxynitrite-specific diazaborine-based chemical tools for imaging and, in the longer term, drug delivery. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.