MiR-409-3p regulates angiogenesis, brown fat adiposity, and insulin resistance

MiR-409-3p 调节血管生成、棕色脂肪肥胖和胰岛素抵抗

• 批准号: 10052898
• 负责人: Basak Icli
• 金额: $ 60.33万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2020-11-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10052898
• 关键词: 3' Untranslated Regions; 3T3-L1 Cells; Ablation; Adipocytes; Adipose tissue; Age; Angiogenesis Inhibition; Angiogenic Factor; Animal Model; Binding; Biological Assay; Blood Vessels; Brown Fat; Cell Proliferation; Coculture Techniques; Code; Development; Diet; Endothelial Cells; Energy Metabolism; Excision; Fatty acid glycerol esters; Functional disorder; Gene Expression; Genes; Growth; Growth Factor; High Fat Diet; Homeobox; Hormones; Human; Impairment; Insulin; Insulin Resistance; Knock-out; Knockout Mice; Length; MAP Kinase Kinase Kinase; Measures; Mediating; Metabolic; Metabolic Diseases; Metabolic dysfunction; Metabolism; MicroRNAs; Molecular; Mus; Non-Insulin-Dependent Diabetes Mellitus; Nucleotides; Nutrient; Obese Mice; Obesity; Organ; Organoids; PECAM1 gene; PGF gene; Patients; Phenocopy; Phosphotransferases; Plasma; Play; Proteins; Regulation; Reporter; Role; Sampling; Signal Pathway; Signal Transduction; Skin; Small Interfering RNA; Stains; Stimulus; Temperature; Testing; Therapeutic Intervention; Thermogenesis; Tissues; Untranslated RNA; Vascular Endothelial Growth Factors; Vascularization; Waste Products; Zinc Fingers; angiogenesis; base; cadherin 5; cell growth; cell motility; conditional knockout; diabetic; disorder risk; endothelial dysfunction; glucose tolerance; improved; in vivo; inhibitor/antagonist; insight; insulin sensitivity; insulin tolerance; knock-down; miRNA expression profiling; mouse model; non-diabetic; obesity development; overexpression; programs; promoter; response; subcutaneous; transcriptome sequencing; uncoupling protein 1

PROJECT SUMMARY/ABSTRACT White and brown adipose tissues are highly vascularized organs, capable of plasticity based on metabolic demands and energy expenditure. Critical gaps remain in our understanding of how angiogenesis impacts adipose tissue dysfunction and overall metabolism. MicroRNAs (miRs) are implicated in the regulation of the angiogenic response to pathophysiological stimuli. However, the role of miRs in regulating the angiogenic response in diet-induced insulin resistance is poorly understood. Using a miRNA-Seq approach, we identified that miR-409-3p expression was significantly increased in endothelial cells (ECs) of brown adipose tissue (BAT) of diet-induced obese (DIO) mice and in human diabetic plasma samples compared to non-diabetic patients. Overexpression of miR-409-3p markedly inhibited EC growth and migration, whereas miR-409-3p inhibition had the opposite effects. Preliminary studies indicate that miR-409-3p targets the 3'UTRs of Zinc Finger E-box binding Homeobox 1 (ZEB1) and Mitogen-activated protein kinase kinase kinase kinase 3 (MAP4K3). SiRNA knockdown of ZEB1 or MAP4K3 expression in ECs phenocopied the effects of miR-409-3p overexpression and significantly decreased EC proliferation and migration. 3T3-L1 cells or human skin fat organoids co-cultured with conditioned media from ECs overexpressing miR-409-3p had decreased expression of BAT markers (UCP1, Cidea), whereas conditioned media from ECs deficient in miR-409-3p markedly increased expression of BAT gene expression. Mechanistically, Placental Growth Factor (PLGF), secreted from ECs in response to miR-409-3p inhibition promoted 3T3-L1 differentiation towards brown-like adipocytes as measured by increased UCP1 expression. Systemic delivery of LNA-anti-miR-409-3p inhibitor to DIO mice significantly increased angiogenesis as measured by CD31 staining, UCP-1 expression in BAT and subcutaneous white adipose tissue (sWAT), while improving glucose and insulin tolerance and energy expenditure. Therefore, we hypothesize that miR-409-3p serves as a critical regulator of EC growth and angiogenesis in adipose tissue and may improve metabolic dysfunction in DIO. Thus, in Aim1 we will investigate the molecular mechanisms by which miR-409-3p regulates EC growth and angiogenesis. In Aim2, we will delineate the mechanisms by which miR-409-3p in ECs regulates browning in adipose tissues. Finally, in Aim3, we will explore the effect of miR-409-3p neutralization in the vasculature of adipose tissues and development of DIO and insulin resistance in mice. Successful completion of these studies will shed insights on the regulatory role of miR-409-3p between impaired angiogenesis in diet-induced obesity and adipose tissue dysfunction, an effect that could be exploited for therapeutic intervention in obesity-induced insulin resistance. !

项目摘要/摘要 白色和棕色脂肪组织是高度血管化器官，能够基于可塑性 代谢需求和能源消耗。关键的差距在我们对血管生成的理解中仍然存在 影响脂肪组织功能障碍和总体代谢。 microRNA（mir）与调节有关 对病理生理刺激的血管生成反应。但是，mir在调节 饮食诱导的胰岛素抵抗中的血管生成反应知之甚少。 使用miRNA-seq方法，我们确定miR-409-3p表达在 饮食诱导的肥胖（DIO）小鼠的棕色脂肪组织（BAT）的内皮细胞（EC）和人类糖尿病 与非糖尿病患者相比，血浆样品。 miR-409-3p的过表达明显抑制EC 生长和迁移，而miR-409-3p抑制作用具有相反的影响。初步研究表明 miR-409-3p靶向3'UTRS的3'UTRS锌指EBOX结合同源物1（ZEB1）和有丝分裂原激活 蛋白激酶激酶激酶激酶3（MAP4K3）。 EC中Zeb1或Map4K3表达的siRNA敲低 MiR-409-3p过表达的作用，显着降低了EC的增殖和 迁移。 3T3-L1细胞或人类皮肤脂肪器官与EC的条件培养基共培养 过表达miR-409-3p的蝙蝠标记表达降低（UCP1，CIDEA），而条件 来自MiR-409-3p缺乏EC的培养基显着增加了BAT基因表达的表达。 从机械上讲，胎盘生长因子（PLGF），从EC分泌，以响应miR-409-3p抑制作用 通过增加的UCP1表达测量，促进了向棕色脂肪细胞的3T3-L1分化。 LNA-ANTI-MIR-409-3P抑制剂到DIO小鼠的全身递送显着增加了血管生成 通过CD31染色测量，在BAT和皮下白色脂肪组织（SWAT）中表达UCP-1，而 改善葡萄糖和胰岛素耐受性和能量消耗。因此，我们假设miR-409-3p 是脂肪组织中EC生长和血管生成的关键调节剂，可以改善代谢 DIO功能障碍。因此，在AIM1中，我们将研究miR-409-3p的分子机制 调节EC生长和血管生成。在AIM2中，我们将描述mir-409-3p的机制 EC在脂肪组织中调节褐变。最后，在AIM3中，我们将探索miR-409-3p的效果 小鼠脂肪组织脉管系统中和DIO和胰岛素抵抗的发展。 这些研究的成功完成将洞悉miR-409-3p的调节作用 饮食诱导的肥胖和脂肪组织功能障碍中血管生成受损，可以利用这种作用 用于肥胖引起的胰岛素抵抗的治疗干预。 呢