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

项目摘要 细胞衰老，其特征在于稳定的增殖停滞和促炎因子的分泌， 不仅是衰老的标志，也是人类年龄相关疾病的关键因素。随着美国 随着人口老龄化的加剧，更好地了解细胞衰老的紧迫性增加了。 不同的组织。不幸的是，我们仍然缺乏明确定义 衰老在分子和细胞水平，由于其异质性表型。为了解决这一重大问题， 知识的差距，我们建议建立一个组织映射中心，重点是识别和 在健康小鼠脑、骨髓、乳腺、结肠和肝脏中的衰老细胞的表征。我们 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性变化的知识， 组蛋白修饰沿着细胞衰老标记基因的基因表达水平。我们将 部署尖端的单细胞原位和组织分离多组学工具，这些工具已经得到很好的建立 在我们的中心，以产生全面的单细胞分辨率地图的转录组和表观基因组， 雄性和雌性小鼠脑、骨髓、乳腺、结肠和肝脏，并提供定性和 这些重要器官中衰老细胞的正常负荷的定量空间图，在生命周期中， 两种小鼠品系。我们将严格验证新定义的衰老细胞群， 药理学和遗传学方法来根除衰老细胞或抑制它们的炎症 表型，以及用于细胞衰老的正交最新技术水平和常规测定。我们将 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，衰老细胞有益与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综合单细胞图谱将使我们能够识别和 表征不同组织背景和老化过程中的细胞衰老。 我们预计， 这项研究将为未来研究衰老细胞的特征和靶向提供参考 与以下疾病相关或之前的疾病 大脑、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.