Novel in situ proteomics methods to classify cell types in Alzheimer’s brains

用于对阿尔茨海默氏症大脑细胞类型进行分类的新型原位蛋白质组学方法

• 批准号: 9789340
• 负责人: Jia Guo
• 金额: $ 33.26万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-20 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9789340
• 关键词: Address; Adopted; Alzheimer' s Disease; Antibodies; Biological Assay; Biological Markers; Biology; Brain; Brain Diseases; Cells; Chemicals; Clinical; Complex; Detection; Diagnosis; Disease; Dreams; Embryo; Epitopes; Excision; Fluorescence; Fluorescent Antibody Technique; Formalin; Gene Expression Regulation; Goals; Hippocampus (Brain); Human; Image; In Situ; Individual; Knowledge; Label; Malignant Neoplasms; Medicine; Methods; Molecular; Molecular Diagnosis; Monitor; Neurobiology; Neurodegenerative Disorders; Organism; Paraffin Embedding; Pathology; Patients; Positioning Attribute; Process; Proteins; Proteomics; Sampling; Signal Transduction; Solid Neoplasm; Stains; Technology; Time; Tissue Embedding; Tissues; antibody conjugate; base; biomaterial compatibility; brain cell; brain tissue; cancer cell; cell type; cellular targeting; chemical cleavage; fluorescence imaging; imaging modality; improved; mental function; new therapeutic target; novel; preservation; prevent; protein profiling; stem cell biology; targeted treatment

Project Summary Comprehensive in situ protein profiling in individual cells of intact tissues holds great promise to unlock major mysteries in biology and medicine, since it can reveal the spatial organization, gene expression regulation, and interactions of the diverse cell types in multicellular organisms, such as brain tissues, solid tumors and developing embryos. However, this dream remains largely unrealized as the current multiplexed protein imaging methods are limited by their low detection sensitivity, sample throughput and multiplexing capacity. These fundamental limitations of the existing methods hinder their applications to profile formalin- fixed paraffin-embedded (FFPE) tissues, which are the most common type of preserved clinical samples. Our overarching goal is to develop a highly sensitive and high-throughput approach for comprehensive protein profiling in FFPE tissues in situ. To achieve this goal, we propose the following three specific aims. Aim 1: develop cleavable fluorescent antibodies for high-throughput and highly multiplexed protein profiling in FFPE tissues. Aim 2: develop highly sensitive probes for comprehensive protein profiling in FFPE tissues. Aim 3: classify the diverse cell types in human normal and Alzheimer’s brains by single-cell comprehensive protein profiling. If successful, these cleavable probes based technologies will have broad impact on understanding, diagnosis and treatment of complex diseases, such as neurodegenerative diseases and cancer. By mapping regulatory networks within individual cells and interrogating interactions among cells in a tissue, we will enhance our understanding of the molecular mechanisms of diseases, and may discover novel drug targets for more effective cellular targeted therapy. By pinpointing alterations in the abundances and positions of various proteins in single cells, new biomarkers can be identified to transform molecular diagnosis and treatment monitoring. The methods developed in the project can be readily adopted by other labs, and will be made publicly available.

项目摘要 完整的原位蛋白质分析完整组织的单个细胞中有很大的希望解锁 生物学和医学的主要奥秘，因为它可以揭示空间组织，基因表达 调节以及多细胞生物中潜水细胞类型的相互作用，例如脑组织，固体 肿瘤和发展胚胎。但是，随着当前的多路复用，这个梦想在很大程度上仍未实现 蛋白质成像方法受其低检测灵敏度，样品吞吐量和多重限制 容量。现有方法的这些基本局限性阻碍了其应用于概述formorin- 固定的石蜡包裹（FFPE）组织，这是保留的最常见类型的临床样品。我们的 总体目标是开发一种高度敏感和高通量的方法来综合蛋白质 在原位进行FFPE组织分析。为了实现这一目标，我们提出以下三个特定目标。目标1： 在FFPE中开发可裂解的荧光抗体，用于高通量和高度多路复用的蛋白质分析 组织。 AIM 2：为FFPE组织中的全面蛋白质分析发展高度敏感的问题。目标3： 通过单细胞全面蛋白 分析。如果成功，这些基于探测器的技术将对理解有广泛的影响， 复杂疾病的诊断和治疗，例如神经退行性疾病和癌症。通过映射 单个细胞内的调节网络并询问组织中细胞之间的相互作用，我们将 增强我们对疾病分子机制的理解，并可能发现新的药物靶标 更有效的细胞靶向治疗。通过查明各种丰度和位置的改变 单个细胞中的蛋白质，可以鉴定出新的生物标志物来改变分子诊断和治疗 监视。项目中开发的方法可以很容易地由其他实验室采用，并将制定 公开可用。