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

在化学系生命过程化学项目的支持下，来自科罗拉多矿业学院的Dylan Domaille正在研究开发新的荧光化合物和材料，以跟踪细胞系统中的过氧亚硝酸盐。过氧亚硝酸盐是一种由活细胞产生的强效氧化剂，可以破坏DNA和蛋白质等生物分子。然而，过氧亚硝酸根是具有挑战性的跟踪在复杂的生物系统中没有其他化合物的干扰，因为它的高反应性和短的寿命。Domaille博士将开发一种新的化学“开关”，选择性地与过氧亚硝酸盐反应。这种化学开关将激活另一种化合物的荧光，产生一类新的荧光探针，可以照亮细胞产生过氧亚硝酸盐的位置和时间。为了实现这些目标，本科生和研究生将学习合成化学，荧光光谱学和活细胞荧光显微镜。在一个专注于科学传播的互补、双管齐下的教育项目中，本科生将学习如何向广泛的受众（包括专业科学家、学生同行、和高中生。本研究项目将继续合成和评价二氮杂硼杂环-用于追踪过氧亚硝酸根的细胞内荧光探针，其分辨率使得在不同细胞室中进行测量成为可能。还将开发新的成像平台，以追踪具有空间保真度的细胞外过氧亚硝酸盐，以追求单细胞细胞外代谢物成像。这些工具将用于了解过氧亚硝酸盐曲线如何响应机械线索（例如，刚度、粘弹性等）存在于活组织中，但在传统的细胞培养中没有被捕获。拟议的工作旨在a）建立二氮杂硼杂环作为过氧亚硝酸盐特异性基序; B）发明新技术用于成像内和细胞外过氧亚硝酸盐;和c）了解机械线索如何影响内和细胞外氧化代谢产物谱。该项目的结果有望为开发用于成像的过氧亚硝酸盐特异性二氮杂硼杂环化合物化学工具奠定基础，并从长远来看，药物输送。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的知识价值和更广泛的影响审查标准进行评估来支持。

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.