Comprehensive Quantitative Profiling of Cellular Alterations Caused by Injury

损伤引起的细胞改变的全面定量分析

• 批准号: 10392403
• 负责人: PRAMOD K DASH
• 金额: $ 63.73万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10392403
• 关键词: Adoption; Anatomy; Animals; Antibodies; Apoptotic; Astrocytes; Atlases; Biological Assay; Biological Models; Biology; Blood Vessels; Brain; Brain Concussion; Brain Diseases; Brain Injuries; Brain Pathology; Brain region; Cells; Characteristics; Clinical Trials; Communities; Complex; Computer software; Computers; Cortical Cell Layer; Data; Dementia; Development; Disease; Distant; Drug Combinations; Drug Side Effects; Drug usage; Endothelial Cells; Engineering; Failure; Goals; Image; Image Analysis; Individual; Inflammation; Injury; Intelligence; Internet; Investigation; Laboratories; Lighting; Measurement; Methods; Microglia; Modification; Molecular; Morphology; Neurons; Oligodendroglia; Outcome; Pathologic; Pathology; Pharmacotherapy; Photobleaching; Process; Proliferating; Proteins; Protocols documentation; Rattus; Reproducibility; Research; Research Personnel; Research Project Grants; Resolution; Rest; Slice; Software Engineering; Speed; Stains; System; Testing; Therapeutic Intervention; Tissues; Treatment Protocols; Validation; Vascular Endothelial Cell; Visual; Work; base; brain cell; cell type; cost; data acquisition; data mining; design; digital; effective therapy; effectiveness evaluation; excitatory neuron; flexibility; fluorescence microscope; fluorophore; functional status; high resolution imaging; improved outcome; inhibitory neuron; mild traumatic brain injury; next generation; novel drug combination; novel therapeutics; open source; regenerative; side effect; software systems; stem cells; tool; treatment effect; whole slide imaging; Adoption; Anatomy; Animals; Antibodies; Apoptotic; Astrocytes; Atlases; Biological Assay; Biological Models; Biology; Blood Vessels; Brain; Brain Concussion; Brain Diseases; Brain Injuries; Brain Pathology; Brain region; Cells; Characteristics; Clinical Trials; Communities; Complex; Computer software; Computers; Cortical Cell Layer; Data; Dementia; Development; Disease; Distant; Drug Combinations; Drug Side Effects; Drug usage; Endothelial Cells; Engineering; Failure; Goals; Image; Image Analysis; Individual; Inflammation; Injury; Intelligence; Internet; Investigation; Laboratories; Lighting; Measurement; Methods; Microglia; Modification; Molecular; Morphology; Neurons; Oligodendroglia; Outcome; Pathologic; Pathology; Pharmacotherapy; Photobleaching; Process; Proliferating; Proteins; Protocols documentation; Rattus; Reproducibility; Research; Research Personnel; Research Project Grants; Resolution; Rest; Slice; Software Engineering; Speed; Stains; System; Testing; Therapeutic Intervention; Tissues; Treatment Protocols; Validation; Vascular Endothelial Cell; Visual; Work; base; brain cell; cell type; cost; data acquisition; data mining; design; digital; effective therapy; effectiveness evaluation; excitatory neuron; flexibility; fluorescence microscope; fluorophore; functional status; high resolution imaging; improved outcome; inhibitory neuron; mild traumatic brain injury; next generation; novel drug combination; novel therapeutics; open source; regenerative; side effect; software systems; stem cells; tool; treatment effect; whole slide imaging

ABSTRACT Currently, cellular alterations associated with pathological conditions are studied using low complexity immunohistochemical (IHC) assays, typically utilizing 2-5 antibodies, that only reveal a tiny subset of the alterations that are occurring, lack comprehensive cellular context, and do not provide quantitative readouts of cellular changes throughout the tissue. For example, a injury or disease can initiate a complex web of pathological alterations across cell types, and at multiple scales ranging from individual cells to multi-cellular units and the layered brain cytoarchitecture. However, technological limitations are hindering a more comprehensive global understanding of these pathological changes. This lack of understanding is hampering our ability to intelligently design effective treatment regimens, and may have contributed to the failures of clinical trials that targeted a single cell type or specific protein. To bridge this gap in our understanding, we propose to develop a Comprehensive Brain Cellular Alteration Profiling Toolkit (CBAT), a carefully validated and broadly applicable image analysis toolkit with unprecedented potential to accelerate investigation & development of next-generation treatments for brain diseases. CBAT, in association with a flexible and modular protocol for highly multiplexed IHC, will enable simultaneous profiling of all major brain cell types and their functional/pathological status (e.g., resting, reactive, apoptotic) across whole brain sections. It will provide quantitative readouts of cellular alterations at multiple scales ranging from individual cells of all types to multi- cellular units (e.g. niches), brain cell layers, and brain regions. Comprehensive cellular profiling and measurements generated using CBAT will enable a deeper understanding of pathological cellular changes that will enable accelerated design, testing, and optimization of therapeutic interventions. Further, it will reduce overall experimental costs by replacing a large number of less-informative assays with a single comprehensive assay. In the longer term, it will enhance our ability to conduct the systems-level investigations that will be required for fully understanding, and successfully treating, multiple brain pathologies. To achieve these goals, we propose the following aims: Aim 1: Develop and validate a flexible, scalable, extensible, and reproducible method for comprehensive whole slide imaging of all the major brain cell types in stereotactically aligned rat whole brain sections; Aim 2: Develop and validate a turnkey software system profiling cell identify and status at multiple scales ranging from individual cells to multi-cellular units, brain cell layers, and brain anatomic regions; and Aim 3: Test the utility of the CBAT system to comprehensively profile concussion biology, and assess the effectiveness of a drug combination to reduce newly identified pathologies. After its development and validation, CBAT will be disseminated to the research community at no cost for use in their specific research projects.

抽象的 当前，使用低复杂性研究了与病理条件相关的细胞改变 免疫组织化学（IHC）测定法，通常使用2-5种抗体，仅显示了一小部分 发生的改变，缺乏全面的蜂窝情况，并且不提供定量读数 整个组织中的细胞变化。例如，伤害或疾病可以发起复杂的网络 细胞类型之间的病理变化，以及从单个细胞到多细胞的多个量表 单位和分层的大脑细胞结构。但是，技术限制正在妨碍 全球对这些病理变化的全面理解。缺乏理解正在阻碍 我们智能设计有效治疗方案的能力，并可能导致了失败 针对单细胞类型或特定蛋白质的临床试验。为了弥合我们的理解差距，我们 提议开发全面的脑细胞改变分析工具包（CBAT），仔细的 经过验证且广泛适用的图像分析工具包，具有前所未有的潜力以加速调查 ＆开发用于脑部疾病的下一代治疗方法。 CBAT，与灵活的和 高度多重IHC的模块化协议将同时分析所有主要的脑细胞类型，并且 它们的功能/病理状态（例如，静止，反应性，凋亡）在整个大脑切片中。它将提供 从各种类型的单个细胞到多种细胞到多尺度的细胞改变的定量读数 细胞单位（例如壁细分），脑细胞层和大脑区域。全面的细胞分析和 使用CBAT产生的测量将使对病理细胞变化有更深入的了解 将实现治疗干预措施的加速设计，测试和优化。此外，它将减少 总体实验成本通过替换大量不太信息的测定法 测定。从长远来看，它将增强我们进行系统级调查的能力 完全理解并成功治疗多种脑病理所必需的。为了实现这些目标， 我们提出以下目的：目标1：开发和验证灵活，可扩展，可扩展且可重复的 立体定位对准大鼠中所有主要脑细胞类型的全面幻灯片成像的全面幻灯片成像的方法 整个大脑部分；目标2：开发和验证交钥匙软件系统分析单元格识别和状态 从单个细胞到多细胞单元，脑细胞层和脑解剖学的多个量表 地区；目标3：测试CBAT系统的实用性，以全面介绍脑震荡生物学，并 评估药物组合的有效性以减少新鉴定的病理。在其发展之后 和验证，CBAT将无需用于其特定的特定费用，将CBAT分发给研究界 研究项目。