Single nuclei RNA-sequencing to map adipose cellular populations and senescent cells in older subjects

单核 RNA 测序绘制老年受试者的脂肪细胞群和衰老细胞图谱

• 批准号: 10815427
• 负责人: Nicolas Musi
• 金额: $ 68.47万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-15 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10815427
• 关键词: Adipocytes; Adipose tissue; Adult; Affect; Age; Aging; Alzheimer' s Disease; Atherosclerosis; Atlases; Biology of Aging; Cell Aging; Cell Cycle Arrest; Cell physiology; Cell surface; Cells; Dasatinib; Disease; Endothelial Cells; Etiology; Fatty acid glycerol esters; Functional disorder; Global Change; Health; Human; Immune; Individual; Inflammation; Intervention; Laboratories; Malignant Neoplasms; Maps; Mesenchymal Stem Cells; Metabolic; Metabolic Diseases; Metabolic dysfunction; Methodology; Methods; Molecular; Molecular Profiling; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Outcome; Participant; Pharmaceutical Preparations; Phenotype; Play; Population; Property; Quercetin; Research; Resolution; Role; Stromal Cells; Subgroup; Testing; Thinness; Tissues; Variant; bioinformatics tool; cohort; cytotoxicity; glucose tolerance; improved; innovation; insight; insulin sensitivity; lifestyle intervention; novel; novel strategies; obese person; preservation; response; senescence; single nucleus RNA-sequencing; single-cell RNA sequencing; stem; theories; tool; transcriptome

Aging is characterized by an expansion in adipose tissue (i.e. obesity), which plays a key role in the pathophysiology of many aging-diseases, including type 2 diabetes, atherosclerotic vascular disease, Alzheimer’s disease, and some cancers. Adipose tissue is composed of adipocytes, immune cells, endothelial cells, stem/stromal cells, and previously uncharacterized cellular populations. Adipose tissue also contains cells that have undergone cellular senescence, a state characterized by the arrest of the cell cycle and a senescence-associated secretory phenotype (SASP), which induces inflammation, cytotoxicity, and metabolic dysfunction in other cells and tissues. Recent research suggests that aging is associated with changes in adipose cellular populations and subpopulations, and that these variations may be involved in the biology of aging and etiology of aging-related diseases. Yet, the precise profiling of adipose tissue cellular composition has been limited due to a lack of robust cell surface markers for the numerous cellular subpopulations. Single-cell (sc) RNA seq is a novel high-resolution methodology that enables global identification of cellular populations and subpopulations (known and newly identified) in tissues. We have developed a single nuclei (sn)RNA-seq based method (a variation of scRNA-seq) that allows excellent identification, separation and clustering of adipose cellular populations, including mature adipocytes, immune cells, endothelial cells, and mesenchymal stem cells. Although snRNA-seq also is a promising tool for identification and characterization of cells undergoing senescence, it has not been used to more fully elucidate the importance of such cells in the biology of aging and the etiology of aging-related metabolic diseases. In Aim 1 we will conduct adipose tissue molecular profiling via snRNA-seq in three cohorts at baseline: younger lean, older lean, and older obese individuals. We will develop molecular atlases in these subjects to test the hypothesis that both aging and aging plus obesity will have distinct adipose tissue molecular profiles. In a subgroup of older obese subjects we will also test whether a lifestyle intervention changes the global molecular profile in adipose tissue to a profile closer to that in younger and older lean individuals. In Aim 2 we will focus on the identification, quantification, and functional characterization of adipose tissue senescent cells. Sub Aim 2A will test the hypotheses that the amount and function (i.e. SASP) of adipose tissue senescent cells will be elevated with aging and further enhanced by obesity, and that the lifestyle intervention (from Aim 1) will reduce senescent cell burden in older obese subjects. In Sub Aim 2B we will administer senolytic agents to a subgroup of older obese subjects (from Aim 1) to determine whether clearance of senescent cells improves metabolic outcomes. This Sub Aim will directly test the cellular senescence theory and will serve as a positive control for the lifestyle intervention regarding its potential anti-senescence effect.

衰老的特征是脂肪组织的扩张（即肥胖症），在 许多衰老的病理生理学，包括2型糖尿病，动脉粥样硬化血管疾病， 阿尔茨海默氏病和一些癌症。脂肪组织由脂肪细胞，免疫细胞，内皮组成 细胞，茎/基质细胞和以前未表征的细胞群体。脂肪组织还包含 经历了细胞感应的细胞，以细胞周期停滞和a为特征的状态 感应相关的秘书表型（SASP），诱导注射，细胞毒性和代谢 其他细胞和组织的功能障碍。最近的研究表明，衰老与变化有关 脂肪细胞群体和亚群，这些变化可能与 与衰老有关的疾病的衰老和病因。然而，脂肪组织细胞组成的精确分析 由于缺乏众多细胞亚群缺乏健壮的细胞表面标记，因此受到限制。 单细胞（SC）RNA SEQ是一种新型的高分辨率方法，可实现细胞的全局鉴定 在组织中的种群和亚群（已知和新鉴定）。我们已经开发了一个核 （SN）基于RNA-Seq的方法（SCRNA-SEQ的变体），该方法允许出色的识别，分离和 脂肪细胞种群的聚类，包括成熟的脂肪细胞，免疫细胞，内皮细胞和 间充质干细胞。尽管SnRNA-Seq也是识别和表征的有前途的工具 经历着感应的细胞，它并未用于更充分地阐明此类细胞在 衰老的生物学和与衰老相关的代谢疾病的病因。 在AIM 1中，我们将在基线的三个队列中通过SNRNA-SEQ进行脂肪组织分子分析： 年轻的瘦，年长的精益和年长的肥胖个人。我们将在这些受试者中开发分子图谱 检验以下假设：衰老和衰老加肥胖症都将具有不同的脂肪组织分子谱。 在一个老年肥胖主题的子组中，我们还将测试生活方式干预是否改变了全球 脂肪组织中的分子谱，在年轻人和年龄较大的个体中更接近概况。 在AIM 2中，我们将专注于脂肪组织的识别，数量和功能表征 感觉细胞。 Sub Aim 2a将测试脂肪的数量和功能（即SASP）的假设 组织感觉细胞将随着衰老而升高，并通过肥胖而进一步增强，并且生活方式 干预（来自AIM 1）将减少老年肥胖受试者的感觉细胞燃烧。在Sub Aim 2B中，我们将 将鼻溶剂剂给老年肥胖受试者的亚组 感觉细胞改善了代谢结果。该子目标将直接测试细胞感应理论 并将作为其潜在的抗味作用的生活方式干预措施的积极控制。