TriState SenNET (Lung and Heart) Tissue Map and Atlas consortium

衰老细胞随着年龄的增长而积累，并且越来越多地与各种与年龄相关的疾病有关。目前， 衰老细胞是基于一组形态学、生物化学和分子生物学特征来操作性地表征的。 特性.然而，衰老不太可能代表一个单一的统一实体。相反，类似于 从研究肿瘤中了解到，细胞类型，结合特定的内源性驱动因素， 衰老，将可能决定给定衰老细胞的独特分子指纹。理解这一 指纹，就像我们对特定肿瘤中特定基因改变的理解一样，将反过来告知预后 和对治疗的反应我们对衰老生物学的理解大多来自实验 动物模型，而我们对人体细胞和组织衰老的理解充其量仍然是不完整的。 类似地，虽然许多外源性应激可以触发培养物中细胞的衰老表型，但是细胞衰老的表型可能是由细胞衰老引起的。 人类衰老的生理相关驱动因素尚不清楚。最后，对于增殖细胞， 衰老的特征是从细胞周期中永久退出，相关的衰老细胞标志物 和关键有丝分裂后细胞的生物学（例如，心肌细胞、神经元）尚未完全建立。开始 解决这些重大的知识差距，三州SenNet组织映射中心（TMC）将分析 人肺和心脏组织，以提供这些器官中衰老细胞群的高分辨率图谱。 凭借匹兹堡大学和卡内基梅隆大学、俄亥俄州州立大学的综合专业知识， 该财团将与美国大学和罗切斯特大学医学院合作， 评估与各种年龄相关疾病有关的两个器官，其中一个主要是有丝分裂后。我们 分析将涉及使用已建立的靶向测定的原位作图，以及高含量的无偏 方法涉及单细胞RNA/ATAC测序、蛋白质组学和空间转录组学。另外作为 作为我们测绘奋进的一部分，三态SenNet TMC将分析发起 衰老的触发以及衰老细胞的后续生物学和表型。这种关系将 使用来自人类肺和心脏的精确切割组织切片进行探测，这些组织切片将被离体扰动， 特定的衰老诱因这一程序将使我们能够提高检测的精度和灵敏度， 肺和心脏中衰老细胞的定位。最后，我们将纯化和分离衰老的和非衰老的， 来自单个供体肺的衰老细胞，并使用这些等基因原代细胞去卷积， 生理学相关的人类衰老的驱动因素，并描绘所观察到的分子基础， 衰老清除疗法的选择性。总之，这种对来自同一组织的组织和组织切片/细胞的组合分析可以用于组织切片/细胞的分析。 个人将提供一个无与伦比的，前所未有的，深入的，高分辨率的衰老细胞图 人口在人类肺部和心脏，定义衰老的生理驱动因素，并提供一个合理的 了解衰老清除疗法的治疗潜力的方法。

Senescent cells accumulate with age and are increasingly linked to a variety of age-related diseases. At present, senescent cells are operationally characterized based on a set of morphological, biochemical, and molecular properties. However, senescence is unlikely to represent a single uniform entity. Rather, analogous to what has been learned from studying tumors, the cell type, in conjunction with the specific endogenous driver of senescence, will likely determine the unique molecular fingerprint of a given senescent cell. Understanding this fingerprint, like our understanding of the specific genetic alterations in a given tumor, will, in turn, inform prognosis and response to therapy. Much of our understanding of the biology of senescence has come from experimental animal models, while our understanding of senescence in human cells and tissues remains, at best, incomplete. Similarly, while a number of exogenous stresses can trigger a senescent phenotype for cells in culture, the physiological relevant drivers of human senescence are not known. Finally, while for proliferating cells, senescence is characterized by a permanent withdrawal from the cell cycle, the relevant senescent cell markers and biology for critical post-mitotic cells (e.g., cardiac myocytes, neurons) is not well established. To begin to address these significant knowledge gaps, the TriState SenNet Tissue Mapping Center (TMC) will analyze human lung and heart tissue to provide a high-resolution map of the senescent cell population in these organs. With the combined expertise of the University of Pittsburgh and Carnegie Mellon University, Ohio State University, and the University of Rochester School of Medicine, this consortium will provide a comprehensive assessment of two organs linked to a variety of age-related diseases, one of which is largely post-mitotic. Our analysis will involve in situ mapping using established targeted assays, as well as high-content unbiased approaches involving single cell RNA/ATAC sequencing, proteomics, and spatial transcriptomics. In addition, as part of our mapping endeavor, the TriState SenNet TMC will analyze the relationship between the initiating trigger for senescence and the subsequent biology and phenotype of the senescent cell. This relationship will be probed using precision cut tissue sections from human lungs and hearts that will be perturbed ex vivo by specific senescent triggers. This procedure will allow us to increase the precision and sensitivity of detection and localization of senescent cells in the lung and heart. Finally, we will purify and isolate senescent and non- senescent cells from an individual donor lung and used these isogenic primary cells to deconvolute the physiologically relevant driver of human senescence and delineate the molecular basis for the observed selectivity of senolytic therapy. Together, this combined analysis of tissue and tissue slices/cells from the same individual will provide an unrivaled, unprecedented, in-depth, high-resolution map of the senescent cell population in the human lung and heart, define the physiological drivers of senescence, and provide a rational approach to understand the therapeutic potential of senolytic therapy.