Spatial mapping senescent cells across the mouse lifespan by multiplex transcriptomics and epigenomics

项目总结 细胞衰老，以稳定的增殖停滞和促炎因子的分泌为特征， 它不仅是衰老的标志，也是人类衰老相关疾病的关键因素。作为美国 随着人口老龄化，更迫切地需要更好地了解 生命周期中不同的组织。不幸的是，我们仍然缺乏明确定义 由于其表型的异质性，在分子和细胞水平上的衰老。要解决这一重大问题 在知识缺口方面，我们建议建立一个组织图谱中心，专注于识别和 健康小鼠脑、骨髓、乳房、结肠和肝脏衰老细胞的特征。我们的 研究策略建立在我们团队开发的单细胞表观基因组学技术的最新进展之上 以及衰老细胞在染色质景观中表现出特征变化的知识 组蛋白修饰与细胞衰老标记基因的基因表达水平。我们会 部署已经成熟的尖端单细胞原位和组织分离多组体工具 在我们的中心制作转录组和表观基因组的全面单细胞分辨率图谱 雄性和雌性小鼠的脑、骨髓、乳房、结肠和肝脏，并提供定性和 这些重要器官中衰老细胞正常负荷的定量空间图，跨越 两个品系的老鼠。我们将严格验证新定义的衰老细胞群 消除衰老细胞或抑制其炎症的药物和遗传方法 表型，以及细胞衰老的正交最新和常规分析方法。我们会 生成全基因组单细胞DNA甲基化数据，将我们的空间图谱与 表观遗传年龄，衰老细胞有益或有害影响的候选预测因子。我们预计 表观基因组和转录组的全面单细胞图谱将使我们能够识别和 描述细胞在不同组织环境和衰老过程中的衰老。 我们预计计划中的 这项研究将为未来寻求表征和靶向衰老细胞的研究提供参考 与疾病有关的或先于疾病的 脑、骨髓、乳房、结肠和肝脏。

PROJECT SUMMARY Cellular senescence, characterized by stable proliferation arrest and secretion of pro-inflammatory factors, is not only a hallmark of aging, but also a key contributor to age-associated diseases in humans. As the US population is aging, there is an added urgency to gain a better understanding of cellular senescence in different tissues over the lifespan. Unfortunately, we still lack the knowledge to unambiguously define senescence at the molecular and cellular levels, due to its heterogeneous phenotypes. To address this major gap in knowledge, we propose to establish a tissue mapping center that focuses on the identification and characterization of senescent cells in healthy mouse brain, bone marrow, breast, colon and liver. Our research strategy builds on recent advances in single cell epigenomics technologies that our team developed and the knowledge that senescent cells exhibit characteristic changes in the chromatin landscapes and histone modifications along with gene expression levels at marker genes of cellular senescence. We will deploy cutting-edge single cell in situ and tissue dissociative multi-omic tools that have been well established in our center to produce comprehensive single cell resolution maps of the transcriptome and epigenome in male and female mouse brain, bone marrow, breast, colon and liver, and to provide qualitative and quantitative spatial maps of the normal burden of senescent cells in these vital organs, across the lifespan of two mouse strains. We will rigorously validate the newly defined senescent cell populations using pharmacologic and genetic approaches to eradicate senescent cells or suppress their inflammatory phenotype, and orthogonal state-of-the-art and conventional assays for cellular senescence. We will generate whole genome single cell DNA methylation data to link our spatial atlas to measurement of epigenetic age, a candidate predictor of beneficial versus detrimental effects of senescent cells. We expect that comprehensive single cell atlases of epigenome and transcriptome will enable us to identify and characterize cellular senescence in different tissue contexts and during aging. We expect that the planned research will provide a reference for future studies that seek to characterize and target senescent cells associated with or preceding disease in brain, bone marrow, breast, colon and liver.