Platform technologies for scalable highly multiplexed proteomic phenotyping of the brain

用于可扩展的高度多重大脑蛋白质组表型分析的平台技术

• 批准号: 10369777
• 负责人: Kwanghun Chung
• 金额: $ 547.48万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-17 至 2024-09-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10369777
• 关键词: 3-Dimensional; ALS pathology; Adopted; Affect; Amplifiers; Amyotrophic Lateral Sclerosis; Animals; Antibodies; Atlases; Bar Codes; Brain; Brain Diseases; Brain Mapping; Callithrix; Cells; Communities; Complex; DNA; Data; Data Set; Databases; Development; Foundations; Functional disorder; Future; Gene Expression; Goals; Histologic; Human; Image; Individual; Informatics; Label; Libraries; Maps; Methods; Molecular; Motor Cortex; Mus; Neurosciences Research; Nomenclature; Online Systems; Ontology; Organ; Pathologic; Phenotype; Proteins; Proteomics; Protocols documentation; Publishing; Research; Research Personnel; Resolution; Sampling; Speed; Standardization; Structure; Technology; Three-dimensional analysis; Tissues; Visualization software; Work; Yin; base; brain cell; brain tissue; cell type; comparative; computerized tools; cost effective; data dissemination; flexibility; imager; imaging platform; in vivo imaging; informatics tool; insight; lens; multidisciplinary; multimodal data; multimodality; nonhuman primate; novel; protein biomarkers; relating to nervous system; single cell sequencing; tool

The complexity of the mammalian brain is unparalleled by any other organs, and understanding its cellular composition and their brain-wide organization is essential to understand the brain functions and dysfunctions. Extensive efforts have been made toward mapping brain cells through various lenses, and have established invaluable databases yielding new insights. However, only a few molecules, or cell-types, or regions per brain have been mapped in non-human primate and human brains, and fail to capture brain-wide complex intercellular relationships within individuals. We here aim to develop versatile and easy-to-adopt platform technologies for highly multiplexed, rapid, scalable proteomic mapping of mammalian brains from mouse, marmoset, to human. We propose to (Aim 1) develop molecular toolkits and platform technologies (ELAST-SABER) for highly multiplexed scalable proteomic phenotyping of mouse, marmoset, human brains. We will develop a library of orthogonal DNA-barcoded antibodies targeting over 100 canonical cell-type markers, compatible with SABER amplification (developed by the group of PI Peng Yin) and ELAST transport (developed by the group of PI Kwanghun Chung). We will also (Aim 2) validate the technology platform and demonstrate its broad utility and scalability by mapping the cellular landscape in the primary motor cortex (M1) of mouse, marmoset, human brains including tissues with amyotrophic lateral sclerosis (ALS) pathology. We will cross-validate our method using the published MERFISH and single-cell sequencing data from BICCN. We will perform cross-species comparative analysis to understand conserved or evolutionarily distinct cellular features of M1. Finally, we will (Aim 3) establish web-based interactive platforms for data dissemination and community use. We will build multimodal brain atlases allowing cross species comparison, and develop neuro-ontology of the mouse, marmoset and human brain. Importantly, we will develop a database built with informatics and online visualization tools that allow people to view, compare, and analyze multiplexed scalable proteomic imaging data across three species. These informatics tools will have the capacity to standardize and accommodate histological, connectivity, gene expression, and in vivo imaging. Users will be able to compare multi-modality data for the “most comparable structures” either based on their neuroanatomic nomenclature, ontology relationship, similarity of their protein markers, or neural connectivity. These tools will become openly available to the community. Our technology platform will enable us to establish the most comprehensive 3D map of the primary motor cortex (M1) of marmoset and human brains including tissues with amyotrophic lateral sclerosis (ALS) pathology to date— providing a foundation for mapping cell types brain-wide in the future and for studying pathological changes in brain disorders. We envision that these scalable tools will empower the research community to interrogate brain structure and function, including complex intercellular relationships at multiple scales.

哺乳动物大脑的复杂性是任何其他器官都无法比拟的， 组成和它们的全脑组织对于理解大脑功能和功能障碍至关重要。 人们已经做出了广泛的努力，通过各种镜头绘制脑细胞，并建立了 产生新见解的宝贵数据库。然而，每个大脑中只有少数分子或细胞类型或区域 已经在非人类灵长类动物和人类大脑中绘制，并且未能捕获全脑复杂的细胞间 个人内部的关系。我们的目标是开发通用且易于采用的平台技术， 从小鼠、绒猴到人类哺乳动物脑的高度多重、快速、可扩展的蛋白质组图谱。 我们建议（目标1）开发分子工具包和平台技术（ELAST-SABER）， 小鼠、绒猴、人脑的多重可扩展蛋白质组表型分析。我们将建立一个图书馆， 正交DNA条形码抗体靶向超过100个典型的细胞类型标志物，与SABER兼容 扩增（由PI Peng Yin小组开发）和ELAST转运（由PI Peng Yin小组开发） Kwanghun Chung）。我们还将（目标2）验证技术平台，并展示其广泛的实用性， 通过绘制小鼠、绒猴、人类的初级运动皮层（M1）中的细胞景观的可扩展性 脑，包括具有肌萎缩性侧索硬化（ALS）病理学的组织。我们将交叉验证我们的方法 使用已发表的MERFISH和来自BICCN的单细胞测序数据。我们将进行跨物种 比较分析，以了解M1的保守或进化上不同的细胞特征。最后我们将 (Aim 3）建立网上互动平台，供数据发布和社区使用。我们将建立 允许跨物种比较的多模式脑图谱，并开发小鼠的神经本体， 绒猴和人脑。重要的是，我们将开发一个数据库， 可视化工具，允许人们查看、比较和分析多路复用的可扩展蛋白质组成像数据 三个物种。这些信息学工具将有能力标准化和适应组织学， 连接性、基因表达和体内成像。用户将能够比较多模态数据， “最可比结构”或者基于它们的神经解剖学命名、本体关系、相似性 蛋白质标记或神经连接。这些工具将向社区开放。我们 技术平台将使我们能够建立最全面的初级运动皮层（M1）3D地图 包括肌萎缩性侧索硬化症（ALS）病理组织的研究- 为将来绘制全脑细胞类型图和研究脑组织病理变化提供了基础。 大脑紊乱我们设想，这些可扩展的工具将使研究界能够询问大脑 结构和功能，包括复杂的细胞间关系在多个尺度。