Single Cell Characterization of Adipose Tissue in the Setting of HIV and Aging

HIV 和衰老背景下脂肪组织的单细胞表征

• 批准号: 10613754
• 负责人: Heidi J Zapata
• 金额: $ 22.66万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10613754
• 关键词: Acute-Phase Proteins; Adipocytes; Adipose tissue; Adult; Affect; Age; Aging; Architecture; Biopsy; Blood Coagulation Factor; Body mass index; Cells; Central obesity; Characteristics; Chronic; Clinical; Complex; Cytometry; Data; Dendritic Cells; Development; Diabetes Mellitus; Disease; Dyslipidemias; Elderly; Endothelial Cells; Epithelial Cells; Evaluation; Fatty acid glycerol esters; Flow Cytometry; Funding; Genetic Transcription; HIV; HIV Infections; HIV Seronegativity; HIV Seropositivity; Hypertension; Image; Immune; Immunologic Receptors; Individual; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory; Insulin Resistance; Interleukin-1; Interleukin-10; Interleukin-18; Interleukin-6; Link; Metabolic Diseases; Metabolic syndrome; Molecular; Obesity; Output; Paraffin Embedding; Pattern; Population; Process; Proteomics; Sampling; Signal Transduction; Source; TNF gene; TimeLine; Tissues; age group; cell community; comorbidity; cytokine; inflammatory milieu; innate immune mechanisms; innate immune pathways; macrophage; metabolic rate; monocyte; pathogen; protein expression; single-cell RNA sequencing; stem cells; systemic inflammatory response; tissue/cell culture; transcriptomics

Project Summary of Supplement: Both the process of aging and HIV-infection are associated with pro-inflammatory environments that are characterized by elevated levels of cytokines and inflammatory mediators. Stimulation of innate immune receptors by both pathogen-associated molecular patterns and damage associated molecular patterns likely is contributing to the systemic inflammation seen with advancing age, and HIV-infection. There is increasing evidence that adipose tissue-derived inflammation likely contributes to this pro-inflammatory environment. Both obesity and redistribution of adipose tissue occur with the process of aging and HIV-infection and are strongly linked to metabolic syndrome. However, the mechanisms whereby adipose tissue contributes to the development of metabolic syndrome are not fully understood. Adipose tissue is a cellular community that includes adipocytes, endothelial cells, pre-adipocytes, and innate immune cells such as macrophages, dendritic cells, and monocytes. The purpose of this proposal is to characterize both immune and non- immune cell populations within adipose tissue using single cell RNA sequencing, in the context of aging, and HIV-infection in order to understand how they contribute to systemic inflammation. In Aim 1, fat pad biopsies will be collected from HIV-positive, and HIV-negative adults in the following age groups (21- 40) and (≥ 60 years). Multicolor flow cytometry and single cell RNA seq will be used to characterize both immune (macrophages, dendritic cells, monocytes) and non-immune cell populations within adipose tissue by evaluation of innate immune pathways, such as the NLRP3 inflammasome, cytokines (IL-1, IL-18, IL-6, TNF- , IL-10 etc.), and transcriptomics of specific cells to evaluate cellular signaling networks and characterize new cell populations. Aim 2 seeks to characterize protein expression in the context of adipose tissue architecture by using CyTOF imaging mass cytometry. The supernatant of adipose tissue cell cultures will also be collected and used to perform proteomic analysis of the secretome of adipose tissue cells in order to better understand the inflammatory output of these cells. Clinical characteristics (e.g., BMI) and co-morbidities (e.g., diabetes) will be evaluated in conjunction with the experimental data. Our findings will allow us to better understand the underlying innate immune mechanisms within specific cell populations that contribute to adipose tissue derived inflammation and ultimately affect the pro-inflammatory environment that is seen in the setting of aging, and HIV-infection. 2

补充项目摘要： 衰老和艾滋病毒感染的过程都与促炎环境有关， 其特征在于细胞因子和炎症介质水平升高。刺激先天免疫 受体的病原体相关的分子模式和损伤相关的分子模式可能是 导致随着年龄增长而出现的全身性炎症和HIV感染。人们日益 证据表明脂肪组织衍生的炎症可能有助于这种促炎环境。两 肥胖和脂肪组织的重新分布随着衰老和HIV感染的过程而发生， 与代谢综合征有关然而，脂肪组织促进脂肪代谢的机制并不清楚。 代谢综合征的发展尚未完全了解。脂肪组织是一个细胞群落， 包括脂肪细胞、内皮细胞、前脂肪细胞和先天免疫细胞如巨噬细胞， 树突细胞和单核细胞。该提案的目的是描述免疫和非免疫的特点， 使用单细胞RNA测序的脂肪组织内的免疫细胞群体， 衰老和艾滋病毒感染，以了解它们如何导致全身炎症。在目标1中， 将从以下年龄组的HIV阳性和HIV阴性成年人中收集脂肪垫活组织检查（21- 40)（≥ 60岁）。多色流式细胞术和单细胞RNA测序将用于表征 免疫细胞（巨噬细胞、树突细胞、单核细胞）和非免疫细胞群， 评估先天免疫途径，例如NLRP 3炎性体、细胞因子（IL-1 β、IL-18、IL-6、TNF-α） IL-10等），和特定细胞的转录组学来评估细胞信号网络和表征新的 细胞群目的2旨在描述脂肪组织结构中蛋白质表达的特征 通过使用CyTOF成像质谱仪。脂肪组织细胞培养物的上清液也将被纯化。 收集并用于进行脂肪组织细胞分泌组的蛋白质组学分析，以便更好地 了解这些细胞的炎症输出。临床特征（例如，BMI）和合并症（例如， 糖尿病）将结合实验数据进行评估。我们的发现将使我们能够更好地 了解特定细胞群内的潜在先天免疫机制， 脂肪组织来源的炎症，并最终影响促炎环境，这是在 衰老和艾滋病感染的环境。 2