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

衰老细胞随着年龄的增长而积累，并越来越多地与各种与年龄相关的疾病有关。目前， 衰老细胞的操作特征是基于一组形态、生化和分子 属性。然而，衰老不太可能代表一个统一的实体。更确切地说，类似于 是从研究肿瘤、细胞类型和特定的内源性驱动因素中学到的 衰老，可能会决定特定衰老细胞的独特分子指纹。理解这一点 指纹，就像我们对特定肿瘤中特定基因变化的理解一样，反过来也会告诉我们预后 对治疗的反应。我们对衰老生物学的大部分了解都来自于实验 动物模型，而我们对人类细胞和组织衰老的理解充其量也是不完整的。 类似地，虽然一些外源压力可以触发培养中细胞的衰老表型，但 人类衰老的生理相关驱动因素尚不清楚。最后，对于增殖的细胞来说， 衰老的特征是永久性地退出细胞周期，相关的衰老细胞标志物 而关键的有丝分裂后细胞(例如，心肌细胞、神经元)的生物学还没有很好的建立。开始，开始 解决这些重要的知识差距，TriState Sennet组织测绘中心(TMC)将分析 人体肺和心脏组织，提供这些器官中衰老细胞群的高分辨率地图。 与匹兹堡大学和俄亥俄州立卡内基梅隆大学的专业知识相结合 大学和罗切斯特大学医学院，该财团将提供全面的 评估与各种年龄相关疾病有关的两个器官，其中一种主要是有丝分裂后疾病。我们的 分析将包括使用已建立的有针对性的分析以及高含量的无偏见的原位测绘 涉及单细胞RNA/ATAC测序、蛋白质组学和空间转录组学的方法。此外，AS 作为我们测绘工作的一部分，TriState Sennet TMC将分析发起人和 触发衰老以及随后衰老细胞的生物学和表型。这种关系将会 使用精密切割的人肺和心脏组织切片进行探测，这些组织切片将在体外受到 特定的衰老诱因。这一程序将使我们能够提高检测的精确度和灵敏度 衰老细胞在肺和心脏的定位。最后，我们将提纯和分离衰老和非衰老的 从单个供体肺中提取衰老细胞，并使用这些同基因的原代细胞去卷曲 与人类衰老相关的生理驱动因素，并描绘了观察到的 裂解疗法的选择性。总而言之，这种对来自同一组织的组织和组织切片/细胞的联合分析 个人将提供无与伦比的、史无前例的、深入的、高分辨率的衰老细胞地图 在人类的肺脏和心脏中，定义了衰老的生理驱动因素，并提供了合理的 了解感觉神经溶解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.