Spatially-resolved protein and transcriptome mapping of senescent cells

项目摘要 该项目的目标是产生一种新的技术管道，以表型衰老细胞的身份（定义）， 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）。使用不同的特征分析 方法已经揭示衰老细胞是高度异质的，并且不利地影响组织健康， 功能然而，由于生物异质性和依赖于不同的方法，我们不全面 了解衰老细胞的特性以及它们对年龄相关衰退的贡献程度。这限制 能够设计出具有重要社会效益的有效疗法。为了填补这些重要的 由于知识上的差距，我们需要新的技术来准确地表征异质衰老细胞状态， 在老化的组织中。我们将开创成像质谱细胞术（IMC）和转录组学的迭代和综合使用， 数字空间分析（DSP）对衰老组织中衰老细胞的分子表型的研究。我们的初步数据 从高维空间分辨和基于悬浮的映射技术产生的演示 小胶质细胞、神经元和其他细胞类型在老年小鼠大脑中显示出不同的衰老特征。 大脑的独特性质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，以及衰老相关生物标志物的多效性表现，所有这些都在明确的微观范围内， 环境.这些特征支持了对大脑进行可行和全面的初步实验重点 解决预期的衰老的身份和背景的全套。因此，我们将开发这种新技术- nology首先在老年小鼠的海马和皮质（UG 3期）中绘制衰老细胞，随后， 在雌性和雄性小鼠的整个生命周期（UH 3期）中，关键是，我们的创新 通过设计，技术管道将通过定制细胞身份而广泛适用于任何组织 和衰老生物标志物。我们将利用研究细胞衰老和SASP的经验， 组织，以协同适应和缩放IMC和DSP，以生成小鼠和人类衰老细胞图谱。 预计高维多标记成像和空间转录组学将彻底改变 严格和全面地表征和映射不同组织环境中的衰老细胞。最终，使用 这项新技术的应用可能会从根本上推进对细胞衰老如何影响年龄的理解- 相关的组织功能障碍，并可能揭示新的策略，破坏衰老介导的病理。

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.