Optimization of imaging mass cytometry, a single-cell spatial proteomics technology, for the study of Alzheimer disease

用于阿尔茨海默病研究的成像质量细胞术（一种单细胞空间蛋白质组学技术）的优化

• 批准号: 10447479
• 负责人: Jinbin Xu
• 金额: $ 43.31万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10447479
• 关键词: Ablation; Affect; Aging; Alzheimer' s Disease; Alzheimer' s disease pathology; Alzheimer' s disease patient; Alzheimer’s disease biomarker; Amyloid beta-Protein; Antibodies; Architecture; Astrocytes; Autopsy; Biological Markers; Biology; Brain; Cell Communication; Cell Nucleus; Cells; Clinical; Collaborations; Communities; Cytometry; Data Analyses; Detection; Development; Diabetes Mellitus; Disease; Disease Progression; Freezing; Gene Expression; Genomics; Heterogeneity; Histology; Human; Image; Imaging technology; Immune; Immunohistochemistry; Immunology; Infectious Diseases Research; Isotope Labeling; Lasers; Link; Location; Malignant Neoplasms; Mass Spectrum Analysis; Metals; Methodology; Microglia; Molecular; Morphology; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurofibrillary Tangles; Neuroglia; Neurons; Neurosciences; Pathogenesis; Physiological; Play; Proteomics; Research; Resolution; Role; Sampling; Senile Plaques; Severity of illness; Spatial Distribution; Stains; Synapses; Technology; Tissues; Treatment outcome; analytical tool; base; brain tissue; case control; cell type; complex biological systems; computational pipelines; computational platform; design; disease prognosis; effective therapy; excitatory neuron; extracellular; frontal lobe; glial activation; high dimensionality; inhibitory neuron; insight; mental function; neuroimaging; neuron loss; neuronal survival; novel; oligodendrocyte precursor; protein biomarkers; single cell technology; therapeutically effective; tool; transcriptome sequencing; treatment strategy; whole genome

Project Summary This proposal aims to fill a critical gap in applying high-dimensional single-cell spatial proteomics technology - imaging mass cytometry (IMC) - to study neurons and glia in the human brains. IMC is an emerging high-plex technology which combines immunohistochemistry (IHC) staining using metal isotope-labeled antibodies with laser ablation and mass-spectrometry-based detection to produce high-dimensional images. IMC allows the detection of 40-100 protein markers simultaneously on tissue sections and enables a variety of distinct cell types to be analyzed concurrently at a spatially-resolved single-cell resolution. IMC has been applied to cancer, diabetes, immunology, and infectious disease research, identifying functionally distinct immune cell subpopulations associated with disease progression, treatment outcomes, and biomarkers for disease prognosis. As the most common neurodegenerative disease without a disease-modifying treatment, the application of IMC to Alzheimer disease (AD) could provide novel insights into the disease pathogenesis and targets for the development of effective therapeutic. This spatially-resolved single-cell resolution proteomic technology is critical for investigating the impact of AD pathology on different cell types in a spatial context. The extracellular beta- amyloid (Aβ) plaques and the intracellular neurofibrillary tangles (NFT) are hallmarks of AD pathology. Aβ plaques have been linked to selective neuronal death in AD and are surrounded by activated astrocytes and microglia. The burden and location of NFT correlate with neuronal loss, disease severity, and clinical course. However, the molecular mechanisms underlying AD pathologies in relationship to cellular and gene expression changes and neurodegeneration in the human brain remain unclear, which significantly hampers the development of effective treatment strategies. The main challenge of applying IMC to AD research is the lack of a well-designed, validated high-quality antibody panel suited for the brain tissue. This proposal will fill this gap by developing a 37-antibody panel for the analysis of snap-frozen human brain tissues using IMC which will include both the markers to identify all the major cell types in the central nervous system and biomarkers of AD pathologies. In Aim 1, we will design and optimize the immunodetection conditions for a 37-antibody panel for use on postmortem human brain tissue sections and apply the technology to the frontal cortex tissues of AD and matched control cases. In Aim 2, we will develop and optimize a computational pipeline for brain tissue-specific IMC data analysis. This technology could be used to obtain comprehensive spatial information of brain microenvironment and cell-cell interactions among the glia and neurons, which will advance our understanding of how the spatial distribution of Aβ plaques and NFTs affect glial activation and neuronal death. This optimized methodology will provide the neuroscience community a powerful IMC neuroimaging tool for a broad range of research fields such as aging, neurodegeneration, and brain development.

项目概要 该提案旨在填补高维单细胞空间蛋白质组学技术应用的关键空白—— 成像质谱流式细胞仪 (IMC) - 用于研究人脑中的神经元和神经胶质细胞，是一种新兴的高复合技术。 该技术将金属同位素标记抗体的免疫组织化学 (IHC) 染色与 激光烧蚀和基于质谱的检测可产生高维图像。 在组织切片上同时检测 40-100 个蛋白质标记物，并支持多种不同的细胞类型 以空间分辨的单细胞分辨率同时进行分析已应用于癌症， 糖尿病、免疫学和传染病研究，识别功能不同的免疫细胞 与疾病进展、治疗结果和疾病预后生物标志物相关的亚群。 作为最常见的没有疾病修饰治疗的神经退行性疾病，IMC 的应用 阿尔茨海默病（AD）的研究可以为疾病发病机制和治疗靶点提供新的见解。 这种空间分辨的单细胞分辨率蛋白质组技术的开发至关重要。 用于研究 AD 病理学对空间环境中不同细胞类型的影响。 淀粉样蛋白 (Aβ) 斑块和细胞内神经原纤维缠结 (NFT) 是 AD Aβ 病理学的标志。 斑块与 AD 中的选择性神经元死亡有关，并且被激活的星形胶质细胞包围 小胶质细胞的负担和位置与神经元损失、疾病严重程度和临床病程相关。 然而，AD 病理学背后与细胞和基因表达相关的分子机制 人脑的变化和神经退行性疾病仍不清楚，这极大地阻碍了 将 IMC 应用到 AD 研究中的主要挑战是缺乏有效的治疗策略。 精心设计、经过验证的适用于脑组织的高质量抗体组合将填补这一空白。 通过开发 37 抗体面板，使用 IMC 分析速冻的人脑组织，这将 包括识别中枢神经系统所有主要细胞类型的标记物和 AD 的生物标记物 在目标 1 中，我们将设计和优化 37 抗体面板的免疫检测条件。 用于死后人类脑组织切片，并将该技术应用于 AD 和 AD 的额叶皮层组织 在目标 2 中，我们将开发和优化针对特定脑组织的计算流程。 IMC数据分析可用于获取大脑的全面空间信息。 神经胶质细胞和神经元之间的微环境和细胞间相互作用，这将增进我们的理解 研究 Aβ 斑块和 NFT 的空间分布如何影响神经胶质激活和神经死亡。 该方法将为神经科学界提供强大的 IMC 神经影像工具，用于广泛的研究 研究领域包括衰老、神经退行性疾病和大脑发育。