Highly sensitive proteomics method to probe cell heterogeneity at single cell resolution

高灵敏度蛋白质组学方法以单细胞分辨率探测细胞异质性

• 批准号: 10449281
• 负责人: Yu Gao
• 金额: $ 39.41万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10449281
• 关键词: Bacteria; Cell Line; Cells; Cellular Structures; Chemicals; Coupled; Data; Detection; Development; Diagnosis; Disease; Event; Evolution; Fostering; Goals; Health; Heterogeneity; Individual; Knowledge; Label; Mammalian Cell; Measures; Messenger RNA; Methods; Mission; Mitochondria; National Institute of General Medical Sciences; Neoplasm Metastasis; Neurodegenerative Disorders; Peptides; Pharmaceutical Preparations; Pharmacotherapy; Phosphorylation; Post-Translational Protein Processing; Prevention; Proteome; Proteomics; Public Health; Research; Research Personnel; Research Support; Resolution; Resource Development; Resources; Scientific Advances and Accomplishments; Signal Transduction; Statistical Models; Strategic Planning; United States National Institutes of Health; developmental neurobiology; driving force; human disease; improved; informatics tool; innovation; insight; ionization; neuron development; new technology; novel; protein expression; transcriptomics; tumor

Project Summary/Abstract: As the single-cell transcriptomics analysis becomes more accessible, individual cells can now be classified individually, allowing us to understand cell heterogeneity as one of the key driving forces in neuron development and tumor evolution. However, the presence of mRNA is not always coupled with protein expression, single-cell proteomics will provide a more precise view of the cell components and signaling, including key post-translational modification events such as phosphorylation. Here we propose to combine several different strategies to develop a highly sensitive proteomics method to probe cell heterogeneity at single cell resolution. Specifically, we will 1) develop chemical labeling probes to enhance peptide ionization; 2) develop an isobaric isotopologue labeling approach to quantify single mammalian cell from various cell lines; 3) develop informatics tools and statistical models to resolve cell heterogeneity at single-cell resolution with non-single-cell proteomics data. Each of these proposed methods alone, could potentially enable us to measure the proteome of a single mammalian cell. When combined, they will result in a highly sensitive platform that could potentially allow us to probe the proteome at the level of a single mitochondria or bacteria. The long-term goal of this project is to elucidate the fundamental mechanisms of the origin and implications of cell heterogeneity. This proposed research is highly relevant to public health in the fields of tumor evolution, cancer metastasis, developmental neurobiology and neurodegenerative disease. Understanding cell heterogeneity will help us to develop more effective drug treatments to many diseases. Therefore, the proposed research is relevant to the part of NIH's mission that fosters “fundamental creative discoveries, innovative research strategies, and their applications as a basis for ultimately protecting and improving health” and that “supports research in the causes, diagnosis, prevention, and cure of human diseases.” The proposed research is relevant to the NIGMS 2015-2020 strategic plan: “Objective 1-1: Invest in and sustain a broad and diverse portfolio of highly meritorious research.”, “Objective 1-2: Promote the ability of investigators to pursue new research directions, novel scientific insights and innovative ideas”, and “Objective 3-1: Support access to essential research resources and the development of new technologies that enable novel scientific advances.”

项目摘要/摘要： 随着单细胞转录组学分析变得更加易于访问，单个单元格 现在可以单独分类，使我们能够理解细胞异质性 神经元发育和肿瘤进化的关键驱动力。但是，存在 mRNA并不总是与蛋白质表达相结合，单细胞蛋白质组学将提供更多 细胞组件和信号传导的精确视图，包括关键的翻译后修改 磷酸化等事件。 在这里，我们建议将几种不同的策略结合起来，以发展高度敏感 单细胞分辨率下探测细胞异质性的蛋白质组学方法。具体来说，我们将1） 开发化学标记问题以增强肽电离； 2）开发同类 从各种细胞系量化单哺乳动物细胞的同位素标记方法； 3） 开发信息工具和统计模型以解决单细胞的细胞异质性 使用非单独细胞蛋白质组学数据的分辨率。仅这些提出的方法都可以 有可能使我们能够测量单个哺乳动物细胞的蛋白质组。当组合时 它们将导致一个高度敏感的平台，有可能使我们能够探究 单一线粒体或细菌水平的蛋白质组。 该项目的长期目标是阐明 细胞异质性的起源和含义。这项拟议的研究与公众高度相关 肿瘤进化，癌症转移，发育神经生物学和 神经退行性疾病。了解细胞异质性将帮助我们发展更多 许多疾病的有效药物治疗。因此，拟议的研究与 NIH的使命是促进“基本创意发现，创新研究的一部分 策略及其作为最终保护和改善健康的基础的应用程序”和 “支持人类疾病的原因，诊断，预防和治愈方面的研究”。 拟议的研究与NIGMS 2015-2020战略计划有关：“目标1-1： 投资并维持广泛而多样化的高度优势研究组合。”，“客观 1-2：促进调查人员购买新研究方向的能力，新型科学 洞察力和创新思想”和“目标3-1：支持访问基本研究 资源和发展新技术的发展，可以实现新的科学进步。”