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

基本信息

批准号：
10468741
负责人：
Jia Guo
金额：
$ 32.73万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-20 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10468741
关键词：
Address Adopted Alzheimer&apos s disease brain Alzheimer&apos s disease pathology Alzheimer&apos s disease patient Antibodies Biological Assay Biological Markers Biology Brain Brain Diseases Cells Chemicals Clinical Complex 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 Positioning Attribute Process Proteins Proteomics Sampling Signal Transduction Solid Neoplasm Stains Technology Time Tissue Embedding Tissues antibody conjugate antibody detection base biomaterial compatibility brain cell brain tissue cancer cell cell type cellular targeting chemical cleavage detection sensitivity fluorescence imaging imaging modality improved mental function new therapeutic target novel preservation prevent protein profiling stem cell biology targeted treatment

项目摘要

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.

项目概要完整组织单个细胞的全面原位蛋白质分析有望解锁生物学和医学的重大奥秘，因为它可以揭示空间组织、基因表达多细胞生物体中不同细胞类型的调节和相互作用，例如脑组织、固体然而，由于目前的多路复用技术，这一梦想在很大程度上仍未实现。蛋白质成像方法受到检测灵敏度低、样品通量和多重检测的限制现有方法的这些基本局限性阻碍了它们在福尔马林分析中的应用。固定石蜡包埋（FFPE）组织，这是保存临床样本的最常见类型。总体目标是开发一种灵敏的、高通量的综合蛋白质分析方法为了实现这一目标，我们提出以下三个具体目标：开发可裂解荧光抗体，用于 FFPE 中的高通量和高度多重蛋白质分析目标 2：开发用于 FFPE 组织中全面蛋白质分析的灵敏高度探针。通过单细胞综合蛋白对人类正常和阿尔茨海默氏症大脑中的多种细胞类型进行分类如果成功，这些基于可裂解探针的技术将对理解产生广泛的影响，通过绘图来诊断和治疗复杂疾病，例如神经退行性疾病和癌症。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点通过精确定位各种丰度和位置的变化，实现更有效的细胞靶向治疗。单细胞中的蛋白质，可以识别新的生物标志物以改变分子诊断和治疗该项目开发的方法可以很容易地被其他实验室采用，并将进行实施。公开可用。