Bioanalysis Core

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 健康雄性和雌性小鼠脑、骨髓、乳腺、结肠和 肝脏我们的研究策略建立在单细胞表观基因组学技术的最新进展之上， 衰老细胞在染色质中表现出特征性变化的知识 景观，组蛋白修饰和细胞衰老标记基因的表达。我们将部署 尖端的单细胞原位和组织解离多组学工具，已在我们的 中心生产的转录组和表观基因组在男性的全面单细胞分辨率地图 和雌性小鼠脑、骨髓、乳腺、结肠和肝脏，并提供定性和定量 这些重要器官中衰老细胞的正常负荷的空间图，在两只小鼠的寿命期间 菌株我们将严格验证新定义的衰老细胞群，使用药理学和生物学方法。 遗传方法根除衰老细胞或抑制其炎症表型，以及正交 用于细胞衰老的现有技术和常规测定。我们将产生全基因组单细胞 DNA甲基化数据将我们的空间图谱与表观遗传年龄的测量联系起来，表观遗传年龄是一个候选的预测因子。 衰老细胞的有益和有害影响。我们希望全面的单细胞图谱， 表观基因组和转录组将使我们能够识别和表征细胞衰老的不同 组织环境和老化过程中。 我们期望计划中的研究将为未来提供参考 这些研究试图表征和靶向与疾病相关或之前的衰老细胞， 大脑， 骨髓乳腺结肠和肝脏

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 male and female 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, histone modifications and expression of 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.