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

基本信息

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
项目类别：
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

项目摘要

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

项目概要补充：衰老过程和艾滋病毒感染都与促炎症环境有关，其特征是细胞因子和炎症介质水平升高。病原体相关分子模式和损伤相关分子模式的受体可能是随着年龄的增长，导致全身炎症和艾滋病毒感染的情况不断增加。有证据表明脂肪组织来源的炎症可能导致这种促炎症环境。肥胖和脂肪组织的重新分布是随着衰老和艾滋病毒感染的过程而发生的，并且与然而，脂肪组织导致代谢综合征的机制。代谢综合征的发展尚不完全清楚。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，例如巨噬细胞，该提案的目的是表征免疫细胞和非细胞。使用单细胞 RNA 测序分析脂肪组织内的免疫细胞群衰老和艾滋病毒感染，以了解它们如何导致全身炎症。脂肪垫活检将从以下年龄组的 HIV 阳性和 HIV 阴性成年人中采集（21- 40) 和 (≥ 60 年) 将使用多色流式细胞术和单细胞 RNA seq 来表征两者。脂肪组织内的免疫细胞（巨噬细胞、树突状细胞、单核细胞）和非免疫细胞群先天免疫途径的评估，例如 NLRP3 炎性体、细胞因子（IL-1、IL-18、IL-6、TNF- 、IL-10 等）和特定细胞的转录组学，以评估细胞信号网络并表征新的目标 2 旨在表征脂肪组织结构中的蛋白质表达。通过使用 CyTOF 成像质谱流式细胞术，脂肪组织细胞培养物的上清液也将被分离。收集并用于对脂肪组织细胞的分泌组进行蛋白质组学分析，以便更好地了解这些细胞的炎症输出（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进行评估。了解特定细胞群中潜在的先天免疫机制，这些机制有助于脂肪组织衍生的炎症并最终影响促炎环境老龄化和艾滋病毒感染的环境。 2