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

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.

摘要 目前，与病理条件相关的细胞改变是使用低复杂性研究的。 免疫组织化学(IHC)分析，通常利用2-5个抗体，只显示一小部分 正在发生的变化，缺乏全面的细胞背景，并且没有提供定量的读数 整个组织的细胞变化。例如，一次伤害或疾病可能会引发一系列复杂的 不同细胞类型的病理改变，以及从单个细胞到多细胞的多个尺度 单位和分层的大脑细胞结构。然而，技术限制正在阻碍更多的 对这些病理变化的全面全球了解。这种缺乏理解的情况正在阻碍 我们智能设计有效治疗方案的能力，可能是导致 针对单一细胞类型或特定蛋白质的临床试验。为了弥合我们在理解上的差距，我们 建议开发一个全面的脑细胞改变图谱工具包(CBAT)，一个仔细的 经过验证且应用广泛的图像分析工具包，具有前所未有的加速调查的潜力 &开发下一代脑部疾病的治疗方法。CBAT，与灵活和 高度多路复用IHC的模块化协议，将允许同时分析所有主要脑细胞类型和 他们的功能/病理状态(例如，静息、反应、凋亡)横跨整个大脑切片。它将提供 从所有类型的单个细胞到多个细胞的多个尺度上的细胞变化的定量读数 细胞单位(如壁龛)、脑细胞层和大脑区域。全面的蜂窝分析和 使用cBAT生成的测量结果将使我们能够更深入地了解 将加快治疗干预措施的设计、测试和优化。此外，它还将减少 将大量信息量较少的分析替换为单一的全面分析，从而实现总体实验成本 化验。从长远来看，它将增强我们进行系统级调查的能力， 全面了解和成功治疗多种脑部疾病所必需的。为了实现这些目标， 我们提出了以下目标：目标1：开发和验证灵活、可伸缩、可扩展和可重现的 立体定向排列大鼠所有主要脑细胞类型的全脑片成像方法 全脑切片.目标2：开发和验证描述细胞识别和状态的交钥匙软件系统 从单个细胞到多细胞单位、脑细胞层和脑解剖的多个尺度 目标3：测试cBAT系统对全面描述脑震荡生物学的效用，以及 评估一种药物组合的有效性，以减少新发现的病理。在其发展之后 和验证，cBAT将免费传播给研究界，用于其特定的 研究项目。