MiR-409-3p Regulates Angiogensis, 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: 10371707
• 负责人: Basak Icli
• 金额: $ 63.59万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10371707
• 关键词: 3' Untranslated Regions; 3T3-L1 Cells; Ablation; Adipocytes; Adipose tissue; Age; Angiogenesis Inhibition; 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; Harvest; High Fat Diet; Homeobox; Hormones; Human; Immunohistochemistry; Impairment; Insulin; Insulin Resistance; Intravenous; Lead; Length; MAP Kinase Kinase Kinase; Measures; Metabolic; Metabolic Diseases; Metabolic dysfunction; Metabolism; MicroRNAs; Molecular; Mus; Non-Insulin-Dependent Diabetes Mellitus; Nucleotides; Nutrient; Obese Mice; Obesity; Organ; Organoids; PECAM1 gene; Patients; Phenocopy; Phosphotransferases; Plasma; Play; Proteins; Regulation; Role; Sampling; Signal Pathway; Signal Transduction; Skin; Small Interfering RNA; Stains; Stimulus; Testing; Therapeutic Intervention; Thermogenesis; Tissues; Untranslated RNA; Vascular Endothelial Growth Factors; Vascularization; Waste Products; Western Blotting; Zinc Fingers; Zinc deficiency; angiogenesis; base; cell growth; cell motility; diabetic; diet-induced obesity; disorder risk; endothelial dysfunction; glucose tolerance; improved; in vivo; inhibitor/antagonist; insight; insulin sensitivity; insulin tolerance; knock-down; locked nucleic acid; miRNA expression profiling; non-diabetic; obesity development; overexpression; programs; response; subcutaneous; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT White and brown adipose tissues are highly vascularized organs, capable of plasticity based on metabolic demands and energy expenditure. However maladaptive regulation of these tissues can lead to insulin resistance. 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. 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). Overexpression of miR-409-3p decreased ZEB1 and MAP4K3 expression in ECs, whereas inhibition had the opposite effect. 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 supernatant harvested from ECs overexpressing miR-409-3p had decreased expression of brown adipocyte markers (UCP1, Cidea) by RT-qPCR and Western blot analyses, whereas supernatant harvested from ECs deficient in miR-409-3p increased expression of brown adipocyte markers. Systemic intravenous delivery of LNA-anti-miR-409-3p inhibitor to DIO mice significantly increased angiogenesis by CD31 staining, accompanied by higher UCP-1 in BAT and sWAT by RT-qPCR, Western blot, and immunohistochemistry analyses, while improving glucose and insulin tolerance and overall metabolism. 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. To explore this, we first propose in Aim1 to 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。但是，这些组织的适应不良调节可能导致 胰岛素抵抗。临界缺口仍然是我们对血管生成如何影响脂肪组织的理解 功能障碍和整体代谢。在血管生成的调节中实现了microRNA（mirs） 对病理生理刺激的反应。 miR在控制饮食诱导的血管生成反应中的作用 胰岛素抵抗知之甚少。 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靶向锌指E盒结合的3'Utrs homeobox 1（ZEB1）和有丝分裂原激活 蛋白激酶激酶激酶激酶激酶激酶激酶3（MAP4K3）。 miR-409-3p的过表达改善了Zeb1和 EC中的MAP4K3表达，而抑制作用具有相反的作用。 Zeb1或 ECS中的Map4K3表达表达了miR-409-3p过表达的影响，并显着 EC扩散和迁移。 3T3-L1细胞或人类皮肤脂肪器官与 从过表达miR-409-3p的EC收获的上清液改善了棕色脂肪细胞的表达 由RT-QPCR和Western Blot分析的标记（UCP1，CIDEA），而从EC收获的上清液 miR-409-3p缺乏增加棕色脂肪细胞标记的表达。全身静脉输送 LNA-ANTI-MIR-409-3P抑制剂对DIO小鼠的CD31染色显着增加了血管生成， 伴随RT-QPCR，Western印迹和免疫组织化学的BAT和SWAT中的UCP-1较高 分析，同时改善葡萄糖和胰岛素耐受性以及总体代谢。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，可能 改善DIO的代谢功能障碍。为了探讨这一点，我们首先提出AIM1研究分子 miR-409-3p调节EC生长和血管生成的机制。在AIM2中，我们将描绘 EC中miR-409-3p调节脂肪组织中褐变的机制。最后，在AIM3中，我们将 探索miR-409-3p谈判在脂肪组织脉管系统中的影响和DIO的发展 和小鼠的胰岛素抵抗。这些研究的成功完成将使有关监管角色的见解 miR-409-3p在饮食诱导的肥胖症和脂肪组织功能障碍中的血管生成受损之间 可以探索肥胖引起的胰岛素抵抗的治疗干预的作用。