Spatially Resolved Dynamics of Molecular Pathology and Intercellular Interactions in Amytrophic Lateral Sclerosis

肌萎缩侧索硬化症分子病理学和细胞间相互作用的空间分辨动力学

• 批准号: 10549338
• 负责人: Christopher Jackson
• 金额: $ 55.69万
• 依托单位: NEW YORK GENOME CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10549338
• 关键词: ALS pathology; ALS patients; Address; Age; Amyotrophic Lateral Sclerosis; Anatomy; Area; Atlases; Autopsy; Bayesian Analysis; Behavior; Biological Markers; Brain; Candidate Disease Gene; Catalogs; Cells; Clinical; Clinical stratification; Communities; Computer Vision Systems; Computing Methodologies; Data; Data Set; Diagnostic; Disease; Fostering; Functional disorder; Gene Expression; Gene Expression Profile; Genes; Goals; Human; Image; In Situ; Indirect Immunofluorescence; Individual; Knowledge; Link; Localized Disease; Lower Extremity; Machine Learning; Maps; Measurement; Measures; Methods; Modality; Molecular; Motor Cortex; Motor Neurons; Multiomic Data; Mus; Neurodegenerative Disorders; Neuronal Dysfunction; Onset of illness; Paralysed; Pathogenesis; Pathology; Pathway interactions; Patients; Phenotype; Primary Lateral Sclerosis; Proteins; Proteome; Proteomics; Publishing; Research Personnel; Resolution; Resources; Sampling; Signal Transduction; Site; Spinal Cord; Symptoms; Testing; Therapeutic; Time; Tissues; Transcript; Variant; associated symptom; burden of illness; cell type; computer framework; data integration; functional disability; graph theory; human study; imaging modality; immune imaging; interest; molecular dynamics; molecular pathology; mouse model; multidimensional data; multimodality; multiple omics; neuron loss; new therapeutic target; novel; novel diagnostics; patient stratification; precision medicine; protein TDP-43; regional difference; single-cell RNA sequencing; sporadic amyotrophic lateral sclerosis; stem; symptomatology; transcriptome; transcriptome sequencing; transcriptomics; treatment strategy

Project Abstract Amyotrophic lateral sclerosis (ALS) is a devastating progressive neurodegenerative disease in which the loss of upper (primary motor cortex, M1) and lower (spinal cord, SC) motor neurons (MNs) ultimately leads to total paralysis. MN loss in ALS involves cell autonomous and non-cell autonomous activities in multiple cell types of the M1 and SC, the organization of which are well understood. However, there remain 4 major gaps in our knowledge: 1) How ALS-associated molecular pathology in the various cell types of the M1 relates to those in the SC; 2) How subpopulations of specific cell types are spatially arranged in these two regions; 3) How subpopulations of different cell types are organized in higher-order ensembles; and 4) How the coordinated behavior of these ensembles relates to disease-associated molecular pathology (e.g., pathognomonic inclusions). Towards addressing these questions, we propose to develop a spatially resolved multi-omics catalog of cellular subpopulations in the M1 and SC of patients with ALS and healthy controls. By using a combination of approaches to simultaneously map the spatial transcriptome and proteome of all interacting cellular subpopulations in these regions, our aim is to elucidate the origins and temporal dynamics of inter- and intra-cellular activities that may reveal novel diagnostic and therapeutic targets for ALS. Our overarching hypothesis is that ALS pathology stems from dysfunctional MN-glial interactions, and that this predictably differs in the M1 and SC in accordance with patient symptomatology. To address this hypothesis, we propose to use spatially resolved transcriptomic and proteomic measurements to study intact human postmortem tissue from patients stratified by clinical presentation (i.e., site of initial symptom presentation, bulbar or lower limb). We have previously implemented Spatial Transcriptomics on mouse and human SC to identify regional differences within subpopulations of various cell types that vary as a function of disease dynamics. Here, we propose to build upon our existing human study, and for the first time, develop a spatially resolved multi-omics dataset at scale and in the context of disease in matched human postmortem M1 and SC samples (Aim 1), to enable simultaneous exploration of upper and lower motor neurons in the context of intact tissue. These data will be directly tied to measures of ALS pathology (e.g., pathognomonic inclusions). To integrate and analyze relationships between data across modalities, we will develop a computational framework for harmonized analysis of multi-modal, multi- omic measures of ALS disease burden (Aim 2). Finally, we will implement highly multiplexed immuno-imaging to validate top gene candidates generated in Aim 1 at a single-cell level in situ (Aim 3). We expect to obtain an unmatched view of cellular interactions in the postmortem ALS M1 and SC, and to be able to directly link such interactions to features of ALS pathology in situ. This will allow us to identify dysregulated signaling that drives upper and lower motor neuron loss and associated symptoms in patients in ALS.

项目摘要 肌萎缩性外侧硬化症（ALS）是一种毁灭性的进行性神经退行性疾病，其中丧失了 上部（主运动皮层，M1）和下部（脊髓，SC）运动神经元（MNS）最终导致总导致 麻痹。 ALS中的MN损失涉及多种细胞类型的细胞自主和非细胞自主活动 M1和SC，其组织的理解很好。但是，我们有4个主要差距 知识：1）M1各种细胞类型中与ALS相关的分子病理如何与之相关 SC; 2）在这两个区域中如何在空间上排列特定细胞类型的亚群； 3）如何 不同细胞类型的亚群在高阶集合中组织； 4）如何协调 这些合奏的行为与疾病相关的分子病理有关（例如，病理学家 包含）。为了解决这些问题，我们建议开发一个空间解决的多摩斯 ALS和健康对照患者的M1和SC中细胞亚群的目录。通过使用 同时绘制所有相互作用的空间转录组和蛋白质组的方法的组合 在这些区域中，我们的细胞亚群是为了阐明间和时间的起源和时间动力学 细胞内活性可能揭示了ALS的新型诊断和治疗靶标。我们的总体 假设是ALS病理源于功能失调的Mn-胶质相互作用，并且这种可预测的有所不同 根据患者症状学在M1和SC中。为了解决这一假设，我们建议使用 从空间分辨的转录组和蛋白质组学测量中，从 通过临床表现分层的患者（即初始症状表现，鳞茎或下肢的部位）。我们有 先前在小鼠和人类SC上实施的空间转录组学，以识别内部的区域差异 各种细胞类型的亚群，这些细胞类型随疾病动力学的函数而变化。在这里，我们建议以 我们现有的人类研究，也是第一次，大规模开发出空间解决的多摩尼克数据集 匹配的人类后M1和SC样本中疾病的背景（AIM 1），以同时进行 在完整组织的背景下探索上和下运动神经元。这些数据将直接与 ALS病理学的度量（例如，病理学夹杂物）。整合和分析之间的关系 跨模式的数据，我们将开发一个计算框架，用于对多模式，多模式的统一分析 ALS疾病负担的OMIC度量（AIM 2）。最后，我们将实施高度多重的免疫成像 验证在AIM 1中在单细胞水平原位生成的顶级基因候选物（AIM 3）。我们希望获得 验尸M1和SC中细胞相互作用的无与伦比的视图，并能够直接链接此类 与原位ALS病理学特征的相互作用。这将使我们能够识别驱动器的失调信号 ALS患者的上和下运动神经元丧失和相关症状。