MiR-150 regulated adiose tissue B cells in obesity

MiR-150 调节肥胖中的脂肪组织 B 细胞

基本信息

批准号：
10357914
负责人：
Anthony T Vella
金额：
$ 41万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-09 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10357914
关键词：
Adipocytes Adipose tissue Antigen Presentation Antigen Presentation Pathway B-Cell Activation B-Lymphocytes Biological Assay Cell Communication Cell physiology Cells Chronic Clinical Research Computational algorithm Data Development Diabetes Mellitus Ectopic Expression Fatty acid glycerol esters Gene Expression Gene Targeting Genes Goals Health Hematopoiesis Homeostasis Immune Immune response Immunoglobulins Immunologics Inflammation Inflammatory Inflammatory Response Insulin Resistance Intervention Knock-out Knowledge Mature B-Lymphocyte Mediating Metabolic Metabolic Diseases Metabolic syndrome MicroRNAs Modeling Molecular Mouse Strains Mus Obesity Organ Outcome Pathway interactions Patients Physiological Population Process Production Publishing Regulation Reporter Reporting Risk Role Small RNA Stress System T-Lymphocyte Testing Therapeutic Thinness Tissues Work adoptive B cell transfer cell behavior cell type diet-induced obesity in vivo knock-down leukemogenesis macrophage mouse model novel obese patients overexpression preclinical study response screening small hairpin RNA stem cells transcriptome transcriptome sequencing translational potential

项目摘要

Adipose tissue B cells are the second most abundant immune cells found in adipose tissue, and they account for more than 20% of the stromal population within adipose tissue during obesity. Pre-clinical and clinical studies have only recently suggested that adipose tissue B cells can modulate adipose tissue function and contribute to the metabolic syndromes suffered during obesity. However, our understanding of the molecular mechanisms underlying adipose tissue B cell function is limited, and this has significantly hampered the potential translational benefit of targeting B cells that dwell in adipose tissue to mitigate obesity induced metabolic disease. The central goal of our project is to define the key regulators for these B cells during obesity. Nevertheless, our new data has opened a novel point of intervention to control the function of these inflammation-promoting B cells through the action of microRNAs that include miR-150. Our data support the central hypothesis suggesting that: Under the stress of obesity, miR-150 is a key regulator of adipose tissue B cell activation where dysregulation of miR-150 facilitates the pro-inflammatory response of adipose tissue B cells and corrupts their ability to interact with other cells in the adipose tissue niche, which results in exacerbated adipose tissue inflammation and insulin resistance. We will critically test this hypothesis in three specific aims by first demonstrating how miR-150 regulates normal B cell function. This will be achieved with the creation of new and unpublished mouse strains that will allow exquisite control of miR-150 action in B cells during different stages of development. Specifically, we will utilize our novel miR-150 B-cell specific knockout and overexpression mice to define how adipose tissue B cells function under lean and obese conditions. In the second aim we will determine the regulatory role of the gene targets of miR-150 in controlling adipose tissue B cell behavior and function. Specifically, miR-150 regulates B cell pathways through specific target genes and we have screened more than 30 predicted genes and successfully identified several B cell specific miR-150 targets. We will utilize ectopic expression and shRNA knockdown to determine their roles in miR-150-mediated ATB activation and subsequent in vivo function in obese adipose tissue. In aim 3 we will test the hypothesis that disruption of miR-150 in adipose tissue B cells interferes with their ability to control cell-to-cell interactions. We will determine the mechanistic regulation of miR-150 during crosstalk with other cell populations including T cells and macrophages derived from adipose tissue and also adipocytes themselves. We have postulated that part of the mechanism used by the pro-inflammatory B cells is through the MHC II pathway, which will be tested under various physiological conditions. Collectively, these three aims will elucidate the critical regulatory mechanism of microRNA-regulated adipose tissue B cells and their contribution in orchestrating the adipose tissue metabolic/immunologic response to obesity.

脂肪组织 B 细胞是脂肪组织中发现的第二丰富的免疫细胞，它们占肥胖期间脂肪组织内基质细胞群的 20% 以上。临床前和临床研究最近才表明，脂肪组织 B 细胞可以调节脂肪组织功能，并导致肥胖期间遭受的代谢综合征。然而，我们对脂肪组织 B 细胞功能的分子机制的了解是有限的，这极大地阻碍了靶向脂肪组织中的 B 细胞来减轻肥胖引起的代谢疾病的潜在转化益处。我们项目的中心目标是确定肥胖期间这些 B 细胞的关键调节因子。尽管如此，我们的新数据开辟了一个新的干预点，通过包括 miR-150 在内的 microRNA 的作用来控制这些促进炎症的 B 细胞的功能。我们的数据支持以下中心假设：在肥胖压力下，miR-150 是脂肪组织 B 细胞激活的关键调节因子，其中 miR-150 的失调会促进脂肪组织 B 细胞的促炎反应，并破坏它们的能力。与脂肪组织生态位中的其他细胞相互作用，导致脂肪组织炎症和胰岛素抵抗加剧。我们将通过首先证明 miR-150 如何调节正常 B 细胞功能，在三个具体目标中严格检验这一假设。这将通过创建新的未发表的小鼠品系来实现，这些小鼠品系将允许在不同发育阶段精确控制 B 细胞中 miR-150 的作用。具体来说，我们将利用新型 miR-150 B 细胞特异性敲除和过表达小鼠来定义脂肪组织 B 细胞在瘦和肥胖条件下如何发挥作用。第二个目标是确定 miR-150 基因靶标在控制脂肪组织 B 细胞行为和功能中的调节作用。具体而言，miR-150通过特定靶基因调节B细胞通路，我们筛选了30多个预测基因，并成功鉴定了多个B细胞特异性miR-150靶点。我们将利用异位表达和 shRNA 敲低来确定它们在 miR-150 介导的 ATB 激活以及随后在肥胖脂肪组织中的体内功能中的作用。在目标 3 中，我们将测试以下假设：脂肪组织 B 细胞中 miR-150 的破坏会干扰其控制细胞间相互作用的能力。我们将确定 miR-150 在与其他细胞群（包括源自脂肪组织的 T 细胞和巨噬细胞以及脂肪细胞本身）串扰期间的机制调节。我们假设促炎B细胞使用的部分机制是通过MHC II途径，该途径将在各种生理条件下进行测试。总的来说，这三个目标将阐明 microRNA 调节的脂肪组织 B 细胞的关键调节机制及其在协调脂肪组织对肥胖的代谢/免疫反应中的贡献。