Transient Metabolite Detection for Single-Cell Metabolomics and Diagnostics

描述(由申请人提供) 摘要：这项工作将开发在单个细胞内定量和实时地空间检测广泛范围的代谢物的能力。很少有技术可以用来测量代谢物浓度，尽管它们是细胞基因组表达的关键指标，而且到目前为止还没有实现空间和瞬时分辨的代谢物通量测量能力。该项目将提供这些能力，使几个领域受益，包括代谢组学、功能基因组学、毒理学、癌症生物学、代谢工程和临床诊断学。传感器将由单壁碳纳米管组成，它发射出对环境高度敏感的光稳定性近红外荧光。纳米管将被与重要代谢物结合的配体功能化，结合事件将通过纳米管发射的光谱变化进行传递。研究小组将把这些纳米管传感器引入活细胞，以回答与癌症生物学和细胞新陈代谢相关的重要问题。与此同时，该团队将致力于快速和实时地检测与临床相关的代谢物，以用于癌症的早期诊断和转移的基准。我们还将重建多路代谢物数据，以进行单细胞代谢组指纹图谱。 公共卫生相关性：代谢物是人体内产生的小分子，未来可能用于在早期阶段检测癌症，更好地了解癌症的生物学，并改进重要药物的开发。该项目开发了使用纳米材料的传感器，以检测单细胞和人体组织中的多种代谢物。

DESCRIPTION (Provided by the applicant) Abstract: This work will develop the capability to spatially detect a wide range of metabolites quantitatively and in real- time within a single cell. Few techniques are available to measure metabolite concentrations, although they are a key indicator of a cell's genomic expression, and the capability of metabolite flux measurements, resolved spatially and transiently, is heretofore unrealized. This project will provide these abilities to benefit several fields, including metabolomics, functional genomics, toxicology, cancer biology, metabolic engineering, and clinical diagnostics. The sensors will consist of single-walled carbon nanotubes which emit photostable near-infrared fluorescence which is highly environmentally sensitive. The nanotubes will be functionalized with ligands which bind to important metabolites and the binding event will be transduced by spectral shifts in the nanotube's emission. The investigator's group will introduce these nanotube sensors into live cells in order to answer important questions relevant to cancer biology and cell metabolism. Concomitantly, the team will work towards the rapid and real-time detection of clinically-relevant metabolites for early cancer diagnosis and the benchmarking of metastasis. We will also re-construct multiplexed metabolite data in order to conduct single- cell metabolomic fingerprinting. Public Health Relevance: Metabolites are small molecules produced in the body which may be used in the future to detect cancer at earlier stages, better understand the biology of cancer, and improve the development of important drugs. This project develops sensors using nanomaterials to detect many classes of metabolites within single cells and in human tissues.