Single-cell proteomic identification of novel markers of senescence

新型衰老标志物的单细胞蛋白质组学鉴定

• 批准号: 10907052
• 负责人: Zhixun Dou
• 金额: $ 90.18万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10907052
• 关键词: 3-Dimensional; Address; Aging; Autophagocytosis; Behavior; Biological Models; Biology; CDKN2A gene; Cell Aging; Cell Cycle Arrest; Cells; Chromatin; Cytolysis; Cytoplasm; Data Analyses; Detection; Disease; Disease Progression; Embryonic Development; Flow Cytometry; Gamma-H2AX; General Hospitals; Goals; Harvest; Heterogeneity; Human; Impairment; Inflammation; Inflammatory; Inflammatory Response; Institutional Review Boards; Intervention; Lamin B1; Lung; Lung diseases; Macrophage; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Massachusetts; Mission; Mus; New England; Nuclear; Pathologic; Pathway interactions; Phase; Population; Procedures; Proteins; Proteome; Proteomics; Protocols documentation; Research Personnel; Resolution; SIRT1 gene; Services; Specimen; Stimulator of Interferon Genes; Structure of parenchyma of lung; Technology; Testing; Tissues; aged; beta-Galactosidase; biomarker identification; biomarker panel; cell type; cohort; data acquisition; detection sensitivity; embryonic stem cell; human disease; human tissue; improved; innovation; invention; new technology; novel; novel marker; protein complex; senescence; single-cell RNA sequencing; stem cell differentiation; telomere; tissue mapping; tissue preparation; tissue regeneration; tool; tumorigenesis; wound healing

PROJECT SUMMARY Cellular senescence is a stable form of cell cycle arrest associated with pro-inflammatory responses. On the one hand, senescent cells are a barrier for tumorigenesis and promote wound healing and embryogenesis. On the other hand, senescent cells accumulate in aged and diseased tissues, where they impair tissue renewal and contribute to inflammation and disease progression. Identification and characterization of senescent cells in human tissues will contribute to our understanding of human diseases. Thus, mapping senescent cells at the 3-dimensional level and single-cell resolution in human tissues is an important biomedical objective. Accurate mapping of senescent cells requires reliable markers to specifically identify senescent cells. Currently the senescence field has limited markers to unambiguously distinguish between senescent cells and cells in other pathological states. In addition, the available markers do not address the heterogeneity of senescent cells in tissues. To overcome these limitations, we propose to employ a novel single-cell proteomic technology to investigate the proteomes of senescent cells at the single-cell level, with the goal to reveal novel markers of senescence which can be used to identify and map senescent cells in human tissues. Our group has recently developed a novel technology termed Single-Cell ProtEomics by Mass Spectrometry (SCoPE2). This platform combines automated cell lysis, improved detection sensitivity, and optimized data acquisition and analyses, allowing detection and quantification of thousands of proteins within a single cell. We have applied this technology to study embryonic stem cell differentiation and macrophage polarization, revealing their heterogeneity and alterations of proteomes at the single-cell level. In this application, we will use the lung as a model system, which accumulate senescent cells that contribute to aging and lung diseases. In the UG3 phase, we will isolate senescent cells from the lungs of naturally aged mice. The isolated senescent cells together with control cells will be subjected to SCoPE2 procedure to quantify their proteomes at the single-cell level, allowing us to create unique signatures for potentially diverse senescent cell populations. This will help discover novel markers of senescent cells that are not possible with traditional technologies. In the UH3 phase, we will apply this technology to human lungs, to validate and to identify new markers of senescence. In aggregate, we will establish new ways of identifying senescent cells that should offer new tools to probe senescent cells in human tissues, facilitating tissue mapping of senescent cells of the SenNet initiative. In addition, this study has the potential to reveal new biology of senescence, addressing the heterogeneity and proteome alterations at the single-cell level.

项目摘要 细胞衰老是与促炎反应有关的细胞周期停滞形式。在一个 手，衰老细胞是肿瘤发生并促进伤口愈合和胚胎发生的障碍。另一方面 手，衰老细胞积聚在老年和患病的组织中，它们会损害组织的更新并有助于 炎症和疾病进展。人类组织中衰老细胞的鉴定和表征将 有助于我们对人类疾病的理解。因此，在3维水平上绘制衰老细胞， 人体组织中的单细胞分辨率是重要的生物医学物镜。 准确的衰老细胞映射需要可靠的标记以明确识别衰老细胞。 目前，衰老场的标记有限，可以明确区分衰老细胞和 其他病理状态中的细胞。此外，可用标记不能解决衰老的异质性 组织中的细胞。为了克服这些限制，我们建议采用一种新型的单细胞蛋白质组学技术 在单细胞水平上研究衰老细胞的蛋白质组织，目的是揭示新的标志物 衰老可用于识别和绘制人体组织中的衰老细胞。 我们的小组最近开发了一种新型技术，称为单细胞蛋白质组学，按质量学 光谱法（Scope2）。该平台结合了自动细胞裂解，提高检测灵敏度，并 优化的数据获取和分析，允许在A内检测和定量数千种蛋白 单细胞。我们已将这项技术应用于研究胚胎干细胞分化和巨噬细胞 极化，揭示了它们在单细胞水平上蛋白质组织的异质性和改变。 在此应用中，我们将使用肺部作为模型系统，该系统会积累衰老细胞 有助于衰老和肺部疾病。在UG3阶段，我们将从自然的肺中分离衰老细胞 老鼠。分离的衰老细胞与对照细胞一起将进行范围2 在单细胞水平上量化其蛋白质组织，使我们能够为潜在的多样化创建独特的签名 衰老细胞群体。这将有助于发现衰老细胞的新颖标记 传统技术。在UH3阶段，我们将将这项技术应用于人类肺，以验证和验证 确定衰老的新标记。总体而言，我们将建立识别衰老细胞的新方法 这应该提供新的工具来探测人体组织中的衰老细胞，从而促进衰老的组织图 Sennet倡议的细胞。此外，这项研究有可能揭示衰老的新生物学， 解决单细胞水平的异质性和蛋白质组改变。