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.

衰老的特点是脂肪组织的扩张（即肥胖），这在 许多衰老疾病的病理生理学，包括 2 型糖尿病、动脉粥样硬化性血管疾病、 阿尔茨海默病和一些癌症 脂肪组织由脂肪细胞、免疫细胞、内皮细胞组成。 细胞、干/基质细胞和以前未表征的细胞群还含有脂肪组织。 已经经历细胞衰老的细胞，这种状态的特征是细胞周期停滞和 衰老相关分泌表型（SASP），可诱导炎症、细胞毒性和代谢 最近的研究表明，衰老与其他细胞和组织的变化有关。 脂肪细胞群和亚群，并且这些变异可能与脂肪细胞的生物学有关 然而，脂肪组织细胞组成的精确分析。 由于缺乏针对众多细胞亚群的可靠的细胞表面标记，该研究受到了限制。 单细胞 (sc) RNA seq 是一种新颖的高分辨率方法，可实现细胞的全局识别 组织中的群体和亚群（已知的和新识别的）我们已经开发出单个细胞核。 基于 (sn)RNA-seq 的方法（scRNA-seq 的变体）可实现出色的识别、分离和 脂肪细胞群的聚集，包括成熟脂肪细胞、免疫细胞、内皮细胞和 尽管 snRNA-seq 也是一种很有前途的鉴定和表征工具。 尽管细胞正在经历衰老，但它尚未被用来更充分地阐明此类细胞在衰老过程中的重要性。 衰老生物学和衰老相关代谢疾病的病因学。 在目标 1 中，我们将在基线时通过 snRNA-seq 对三个队列进行脂肪组织分子分析： 我们将开发这些受试者的分子图谱以 检验以下假设：衰老和衰老加肥胖都会具有不同的脂肪组织分子特征。 在老年肥胖受试者的亚组中，我们还将测试生活方式干预是否会改变全球 脂肪组织中的分子谱更接近年轻和年长瘦人的分子谱。 在目标 2 中，我们将重点关注脂肪组织的识别、定量和功能表征 子目标 2A 将测试脂肪的数量和功能（即 SASP）的假设。 组织衰老细胞会随着进一步衰老而升高，并因肥胖而进一步增强，并且生活方式 干预（来自目标 1）将减少老年肥胖受试者的衰老细胞负担。在子目标 2B 中，我们将减少衰老细胞的负担。 对老年肥胖受试者亚组（来自目标 1）施用 senolytics 药物以确定是否清除 该子目标将直接测试细胞衰老理论。 并将作为生活方式干预潜在抗衰老作用的积极对照。