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

职业：基于重氮硼啉的工具和平台，用于绘制细胞内和细胞外过氧亚硝酸盐

• 批准号: 2238563
• 负责人: Dylan Domaille
• 金额: $ 57.12万
• 依托单位: Colorado School of Mines
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2238563&HistoricalAwards=false
• 关键词: CAREER; Diazaborine; Based; Tools; Platforms

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.

在化学系生命过程化学项目的支持下，科罗拉多矿业学院的Dylan Domaille正在研究新的荧光化合物和材料的开发，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，由于过氧亚硝酸盐的高反应活性和短寿命，在复杂的生物系统中不受其他化合物干扰的跟踪是一个挑战。多梅耶博士将开发一种新的化学“开关”，可以选择性地与过氧亚硝酸盐发生反应。这种化学开关将激活另一种化合物的荧光，从而产生一种新型荧光探针，可以照亮细胞产生过氧亚硝酸盐的地点和时间。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。在一个互补的、双管齐下的、以科学传播为重点的教育项目中，本科生将学习如何向广泛的受众（包括专业科学家、学生同龄人和高中生）展示从科学数据到生物技术的各种信息和想法。该研究项目将致力于合成和评估基于重氮aborin的细胞内荧光探针，用于跟踪过氧亚硝酸盐的分辨率，使不同细胞区室的测量成为可能。新的成像平台也将开发跟踪细胞外过氧亚硝酸盐的空间保真度，以追求单细胞细胞外代谢物成像。这些工具将用于了解活组织中存在但在传统细胞培养中未捕获的机械线索（例如，刚度，粘弹性等）如何改变过氧亚硝酸盐剖面。拟议的工作旨在a)建立重氮aborines作为过氧亚硝酸盐特异性基序；B)发明成像细胞内和细胞外过氧亚硝酸盐的新技术；c)了解机械线索如何影响细胞内和细胞外氧化代谢物谱。该项目的结果有望为开发基于过氧亚硝酸盐特异性重氮aborine的化学成像工具和长期的药物输送工具提供基础。该奖项反映了美国国家科学基金会的法定使命，并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，被认为值得支持。