Spatially-resolved protein and transcriptome mapping of senescent cells

衰老细胞的空间分辨蛋白质和转录组图谱

• 批准号: 10684900
• 负责人: Marissa Joy Schafer
• 金额: $ 47.5万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-05 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10684900
• 关键词: Address; Age; Aging; Apoptotic; Atlases; Biological Markers; Brain; Brain region; Cell Aging; Cells; Cerebellum; Complex; Corpus striatum structure; Cytometry; Data; Dimensions; Disease; Female; Frequencies; Functional disorder; Gene Expression Profile; Goals; Health; Heterogeneity; Hippocampus; Human; Image; Individual; Intervention; Knowledge; Longevity; Maps; Mediating; Metabolic; Metals; Methods; Microglia; Molecular; Morphology; Mus; Neurons; Pathology; Pathway interactions; Phase; Phenotype; Prefrontal Cortex; Process; Property; Proteins; RNA; Regenerative capacity; Resolution; Slice; Standardization; Structure of choroid plexus; Suspensions; Technology; Tissues; Work; age related; aged; antibody conjugate; biological heterogeneity; biomarker panel; body system; brain cell; brain parenchyma; cell type; design; digital; experience; high dimensionality; human tissue; imaging biomarker; innovation; innovative technologies; male; method development; molecular phenotype; new technology; novel; preservation; programs; response; senescence; single cell sequencing; spatial integration; therapeutic development; therapeutically effective; therapy development; transcriptome; transcriptomic profiling; transcriptomics; white matter

PROJECT SUMMARY The goal of this project is to generate a novel technological pipeline to phenotype senescent cell identity (defined by biomarkers, function-related transcriptional profiles, morphology, and microenvironment) and composition (defined by quantity, diversity, and distribution) across any mouse or human tissue. Use of diverse profiling methods has revealed that senescent cells are highly heterogeneous and adversely influence tissue health and function. However, due to biological heterogeneity and reliance on diverse methods, we do not comprehensively understand the identities of senescent cells and extent to which they contribute to age-related decline. This limits the ability to devise effective therapeutics that could have important societal benefit. To fill these significant knowledge gaps, we require new technologies that accurately characterize heterogenous senescent cell states in aged tissues. We will pioneer iterative and integrated use of imaging mass cytometry (IMC) and transcriptomic digital spatial profiling (DSP) to molecularly phenotype senescent cells in aged tissues. Our preliminary data generated from high-dimensional spatially-resolved and suspension-based mapping technologies demonstrate that microglia, neurons, and additional cell types display distinct senescent profiles in the aged mouse brain. Unique properties of the brain include substantial cell and regional heterogeneity, limited regenerative capacity, age-vulnerability, and pleiotropic presentation of senescence-related biomarkers, all within well-defined micro- environments. These features support an initial experimental focus on the brain for feasible and comprehensive resolution of the anticipated panoply of senescent identities and contexts. Thus, we will develop this novel tech- nology to map senescent cells, first, in the hippocampus and cortex of aged mice (UG3 phase) and subsequently, across brain regions in female and male mice throughout the lifespan (UH3 phase). Critically, our innovative technological pipeline, by design, will be broadly applicable to any tissues through customization of cell-identity and senescence biomarkers. We will leverage experience studying cell senescence and the SASP in diverse tissues to collaboratively adapt and scale IMC and DSP to generate mouse and human senescent cell atlases. High-dimensional multimarker imaging and spatial transcriptomics are anticipated to revolutionize the ability to rigorously and comprehensively characterize and map senescent cells in distinct tissue contexts. Ultimately, use of this novel technology may fundamentally advance understanding of how cell senescence contributes to age- related tissue dysfunction and may reveal new strategies to disrupt senescence-mediated pathology.

项目概要 该项目的目标是开发一种新颖的技术管道来对衰老细胞的身份进行表型分析（定义为 通过生物标志物、功能相关的转录谱、形态学和微环境）和组成 （由数量、多样性和分布定义）跨越任何小鼠或人体组织。使用不同的分析 方法表明，衰老细胞具有高度异质性，会对组织健康产生不利影响 功能。然而，由于生物异质性和对不同方法的依赖，我们并没有全面地 了解衰老细胞的特性以及它们对与年龄相关的衰退的影响程度。这限制了 设计出可能产生重要社会效益的有效疗法的能力。为了填补这些重要 知识差距，我们需要能够准确表征异质衰老细胞状态的新技术 在老化的组织中。我们将率先迭代和集成使用成像质量流式细胞仪 (IMC) 和转录组学 数字空间分析（DSP）对衰老组织中的衰老细胞进行分子表型分析。我们的初步数据 由高维空间分辨率和基于悬浮的测绘技术生成表明 小胶质细胞、神经元和其他细胞类型在老年小鼠大脑中表现出独特的衰老特征。 大脑的独特特性包括大量的细胞和区域异质性、有限的再生能力、 年龄脆弱性和衰老相关生物标志物的多效性表现，都在明确的微观范围内 环境。这些特征支持最初的实验重点放在大脑上，以实现可行和全面的研究 解决预期的衰老身份和环境的问题。因此，我们将开发这项新技术 首先，在老年小鼠的海马体和皮层（UG3 期）中绘制衰老细胞图谱，然后， 雌性和雄性小鼠整个生命周期（UH3 阶段）的大脑区域。至关重要的是，我们的创新 通过设计，技术管道将通过细胞身份的定制广泛适用于任何组织 和衰老生物标志物。我们将利用在不同领域研究细胞衰老和 SASP 的经验 组织协作调整和扩展 IMC 和 DSP，以生成小鼠和人类衰老细胞图谱。 高维多标记成像和空间转录组学预计将彻底改变以下能力： 严格、全面地表征和绘制不同组织环境中衰老细胞的图谱。最终，使用 这项新技术可能会从根本上促进对细胞衰老如何影响年龄的理解 相关的组织功能障碍，并可能揭示破坏衰老介导的病理学的新策略。