Spatially-resolved protein and transcriptome mapping of senescent cells

项目总结 该项目的目标是产生一种新的技术管道来识别衰老细胞的表型(定义 通过生物标志物、与功能相关的转录图谱、形态和微环境)和组成 (由数量、多样性和分布定义)在任何老鼠或人类组织中。使用不同的侧写 方法揭示了衰老细胞高度异质性，并对组织健康和 功能。然而，由于生物的异质性和对多样化方法的依赖，我们没有全面地 了解衰老细胞的特性以及它们在多大程度上促进了与年龄相关的衰退。这限制了 设计出可能具有重要社会效益的有效疗法的能力。以填补这些重要的 知识差距，我们需要新的技术来准确地表征不同的衰老细胞状态 在老化的组织中。我们将开创成像质量细胞术(IMC)和转录术的迭代和集成使用 对衰老组织中衰老细胞分子表型的数字空间轮廓分析。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的演示 在衰老的小鼠大脑中，小胶质细胞、神经元和其他细胞类型显示出不同的衰老特征。 大脑的独特特性包括大量的细胞和区域的异质性，有限的再生能力， 年龄易损性和衰老相关生物标记物的多向性呈现，所有这些都在定义明确的微观 环境。这些特征支持了对大脑的初步实验重点，以实现可行性和全面性 对预期的所有老化的身份和背景的解决。因此，我们将开发这项新技术-- 首先，在老年小鼠的海马区和皮质(UG3期)定位衰老细胞，随后， 在雌性和雄性小鼠的整个生命周期(UH3阶段)中的大脑区域。关键的是，我们的创新 根据设计，技术流水线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。我们将利用在不同领域研究细胞衰老和SASP的经验 组织，以协作适应和扩展IMC和数字信号处理器，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技术有望彻底改变 严格和全面地描述和绘制不同组织环境中衰老细胞的特征和图谱。最终，使用 这项新技术可能从根本上提高对细胞衰老如何影响衰老的理解-- 相关的组织功能障碍，并可能揭示破坏衰老介导的病理的新策略。

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.