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

基本信息

批准号：
10615029
负责人：
Basak Icli
金额：
$ 61.09万
依托单位：
TUFTS MEDICAL CENTER
依托单位国家：
美国
项目类别：
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/10615029
关键词：
3&apos Untranslated Regions 3T3-L1 Cells Ablation Acceleration Adipocytes Adipose tissue Age Angiogenesis Inhibition Angiogenesis Inhibitors Animal Model Binding Biological Assay 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 Resistance Intravenous Length MAP3K1 gene 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 Transcript Untranslated RNA Vascular Endothelial Growth Factors Vascularization Waste Products Western Blotting Zinc Fingers Zinc deficiency angiogenesis cell growth cell motility diabetic diet-induced obesity disorder risk endothelial dysfunction glucose tolerance improved in vivo inhibitor insight insulin sensitivity insulin tolerance knock-down locked nucleic acid miRNA expression profiling non-diabetic obesity development overexpression programs response subcutaneous transcriptome sequencing

项目摘要

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.

项目概要/摘要白色和棕色脂肪组织是高度血管化的器官，能够基于然而，这些组织的适应不良调节可能会导致代谢需求和能量消耗。我们对血管生成如何影响脂肪组织的理解仍存在重大差距。 MicroRNA (miR) 与血管生成的调节有关。对病理生理刺激的反应 miR 在调节饮食诱导的血管生成反应中的作用。人们对胰岛素抵抗知之甚少。使用 miRNA-Seq 方法，我们发现 miR-409-3p 表达在饮食诱导肥胖 (DIO) 小鼠和人类糖尿病患者棕色脂肪组织 (BAT) 的内皮细胞 (EC) 与非糖尿病患者的血浆样本相比，miR-409-3p 的过度表达显着抑制 EC。生长和迁移，而 miR-409-3p 抑制则具有相反的作用。 miR-409-3p 靶向锌指 E 盒结合同源盒 1 (ZEB1) 和丝裂原激活的 3’UTR 蛋白激酶激酶激酶激酶 3 (MAP4K3) 的过度表达会降低 ZEB1 和 ECs 中 MAP4K3 的表达，而 ZEB1 或 siRNA 的抑制则产生相反的效果。 EC 中的 MAP4K3 表达反映了 miR-409-3p 过表达的影响，并且显着共培养的 3T3-L1 细胞或人皮肤脂肪类器官的 EC 增殖和迁移减少。从过表达 miR-409-3p 的 EC 中收获的上清液降低了棕色脂肪细胞的表达通过 RT-qPCR 和蛋白质印迹分析标记（UCP1、Cidea），而从 EC 收获上清液 miR-409-3p 缺陷会增加棕色脂肪细胞标志物的表达。通过 CD31 染色，DIO 小鼠的 LNA-抗 miR-409-3p 抑制剂显着增加血管生成， RT-qPCR、Western blot 和免疫组织化学显示 BAT 和 swAT 中 UCP-1 较高分析，同时改善葡萄糖和胰岛素耐受性以及整体代谢。 miR-409-3p 作为脂肪组织中 EC 生长和血管生成的关键调节因子，可能改善 DIO 的代谢功能障碍为了探索这一点，我们首先在 Aim1 中提出研究分子机制。 miR-409-3p 调节 EC 生长和血管生成的机制在 Aim2 中，我们将描述最后，在 Aim3 中，我们将研究 EC 中 miR-409-3p 调节脂肪组织褐变的机制。探讨 miR-409-3p 中和对脂肪组织脉管系统和 DIO 发育的影响小鼠胰岛素抵抗的成功完成将有助于揭示其调节作用。饮食诱导的肥胖中血管生成受损和脂肪组织功能障碍之间的 miR-409-3p 的关系，可以利用这一效应对肥胖引起的胰岛素抵抗进行治疗干预。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

MicroRNA-409-3p/BTG2 signaling axis improves impaired angiogenesis and wound healing in obese mice.

DOI：
10.1096/fj.202302124rr
发表时间：
2024-02-08
期刊：
The FASEB Journal
影响因子：
0
作者：
Furkan Bestepe;George F Ghanem;Colette M Fritsche;James Weston;Sumedha Sahay;Am;a K Mauro;a;Parul S
通讯作者：
Parul S