Protective role of let-7 microRNAs in brain endothelial dysfunction during ischemia/reperfusion injury

let-7 microRNA在缺血/再灌注损伤期间脑内皮功能障碍中的保护作用

• 批准号: 10307621
• 负责人: Slava ROM
• 金额: $ 34.67万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10307621
• 关键词: Acute; Address; Adhesions; Alzheimer' s Disease; Animal Model; Anti-Inflammatory Agents; Antiinflammatory Effect; Atherosclerosis; Attenuated; Bioinformatics; Blood - brain barrier anatomy; Blood Vessels; Brain; Brain Injuries; Brain Ischemia; Brain imaging; CCL2 gene; CXCL10 gene; CXCR3 gene; Cell Adhesion Molecules; Cells; Central Nervous System Diseases; Cephalic; Characteristics; Clinical Trials; Complementary therapies; Development; Endothelial Cells; Endothelium; Epilepsy; Event; Extravasation; FDA approved; Family; Fibrinolytic Agents; Functional disorder; Future; Gene Expression; Genes; Glucose; Heterogeneous-Nuclear Ribonucleoproteins; Human; Hypoxia; IL8 gene; In Vitro; Incidence; Infarction; Infection; Infectious Encephalitis; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Injury; Ischemia; Ischemic Stroke; Knock-out; Lead; Leukocytes; Light; Literature; Maintenance; MicroRNAs; Middle Cerebral Artery Occlusion; Modeling; Morbidity - disease rate; Multiple Sclerosis; Neuraxis; Neurologic; Neurologic Deficit; Neurological outcome; Neutrophil Infiltration; Oxygen; Parkinson Disease; Participant; Pathogenesis; Patients; Pericytes; Permeability; Phase; Physiology; Plasminogen Activator; Play; Prevention; Process; Property; Proteins; Publishing; RANTES; RNA Interference; Rattus; Regulation; Regulator Genes; Reperfusion Injury; Reperfusion Therapy; Reporting; Role; Secondary to; Site; Stroke; System; Technology; Testing; Therapeutic; Tight Junctions; Transfection; Traumatic Brain Injury; Umbilical vein; Vascular Dementia; Vascular Diseases; Work; alternative treatment; attenuation; base; blood-brain barrier permeabilization; brain endothelial cell; cerebral microvasculature; cytokine; deprivation; endothelial dysfunction; experimental study; gene therapy; hnRNP A1; improved; in vitro Model; in vivo; in vivo Model; in vivo monitoring; innovation; intravital microscopy; migration; monocyte; mortality; nervous system disorder; neuroinflammation; neurovascular; novel; overexpression; post stroke; prevent; recruit; response; restoration; stroke model; stroke outcome; systemic inflammatory response; therapeutic development; tool; trafficking; treatment strategy

It has been recognized that blood brain barrier (BBB) dysfunction exists in most neurological diseases, including stroke, multiple sclerosis, Alzheimer's and Parkinson's diseases, brain infections and epilepsy. Inflammation plays a significant role in BBB injury, secondary to pro-inflammatory factors produced in the brain or blood and leukocyte engagement of brain endothelium. Brain microvascular endothelial cells (BMVEC) are active participants and regulators of inflammatory processes at a site of inflammation. Inflammatory responses in brain endothelium involve hundreds of genes whose expression requires fine-tuned regulation. microRNAs (miRNAs) recently emerged as major regulators of gene expression. Very limited information exists about their participation in inflammatory responses caused by ischemia/reperfusion (I/R) events in brain endothelial cells. We propose to investigate the role of miRNAs in brain endothelium using well-established in vitro systems for functional studies of the BBB and imaging of brain microvasculature in an in vivo model of neuroinflammation. Based on our studies of BBB dysfunction during neuroinflammation, we propose that barrier protection is best achieved when the intervening agents possess anti-inflammatory properties and can stabilize tight junctions. Recently we have identified highly modified miRNAs, belonging to the let-7 miRNA family, which are important for endothelial maintenance. let-7 and miR-98 were predicted to target the inflammatory molecules, CCL2, CCL5, IL8 and IP10/CXCL10. Overexpression of let-7 and miR-98 in vitro and in vivo resulted in reduced leukocyte adhesion to and migration across brain endothelium and diminished expression of pro-inflammatory cytokines. In oxygen glucose deprivation (OGD) followed by reperfusion (OGD/R), an in vitro I/R model, overexpression of these miRNAs led to increased BBB tightness, thereby attenuating barrier `leakiness'. Overexpression of these miRNAs resulted in decreased infarct volume and neutrophil infiltration in the brain in tMCAO, an in vivo I/R stroke animal model. In our proposed study, we will test the overexpression or inhibition of selected miRNAs on BBB tightness and leukocyte-endothelial cell engagement (adhesion/migration). Using bioinformatics, we will identify other targets for miRNAs. Next, we will perform miRNA transfection in vivo and monitor how the presence of miRNAs will change leukocyte adhesion/migration in an animal model of stroke. Proposed experiments will provide identification and functional assessment of miRNAs in brain endothelium, and lead to future therapeutic developments for prevention of deleterious effects of ischemia/reperfusion on the brain.

血脑屏障（BBB）功能障碍在大多数神经系统疾病中存在， 包括中风、多发性硬化症、阿尔茨海默氏症和帕金森氏症、脑感染和癫痫。 炎症在BBB损伤中起重要作用，继发于脑中产生的促炎因子 或血液和白细胞参与脑内皮。脑微血管内皮细胞（BMVEC）是 炎症部位炎症过程的积极参与者和调节者。炎症反应 在大脑内皮中，涉及数百个基因，其表达需要微调调节。微rna （miRNAs）最近成为基因表达的主要调节因子。关于他们的信息非常有限。 参与脑内皮细胞缺血/再灌注（I/R）事件引起的炎症反应。 我们建议使用成熟的体外系统研究miRNAs在脑内皮细胞中的作用， 在神经炎症的体内模型中的BBB的功能研究和脑微血管的成像。 基于我们对神经炎症过程中血脑屏障功能障碍的研究，我们认为屏障保护是最好的 当介入剂具有抗炎特性并且可以稳定紧密连接时，可以实现。 最近，我们发现了高度修饰的miRNA，属于let-7 miRNA家族，这是重要的 维持内皮细胞。let-7和miR-98被预测为靶向炎症分子，CCL 2， CCL 5、IL 8和IP 10/CXCL 10。let-7和miR-98在体外和体内的过表达导致细胞凋亡减少。 白细胞粘附和迁移穿过脑内皮，并减少促炎因子的表达 细胞因子在体外I/R模型--氧糖剥夺（OGD）再灌注（OGD/R）中， 这些miRNA的过表达导致BBB紧密性增加，从而减弱屏障“渗漏”。 这些miRNAs的过表达导致脑梗死体积减少和中性粒细胞浸润。 tMCAO，体内I/R卒中动物模型。在我们提出的研究中，我们将测试过表达或抑制 选择的miRNAs对BBB紧密性和白细胞-内皮细胞接合（粘附/迁移）的影响。使用 生物信息学，我们将确定其他目标的miRNA。接下来，我们将在体内进行miRNA转染， 监测miRNAs的存在如何改变中风动物模型中白细胞的粘附/迁移。 拟议的实验将提供脑内皮中miRNA的鉴定和功能评估， 并导致未来的治疗发展，用于预防缺血/再灌注对 个脑袋

项目成果

Hyperglycemia-Driven Neuroinflammation Compromises BBB Leading to Memory Loss in Both Diabetes Mellitus (DM) Type 1 and Type 2 Mouse Models.

• DOI: 10.1007/s12035-018-1195-5
• 发表时间: 2019-03
• 期刊: Molecular neurobiology
• 影响因子: 5.1
• 作者: Rom S;Zuluaga-Ramirez V;Gajghate S;Seliga A;Winfield M;Heldt NA;Kolpakov MA;Bashkirova YV;Sabri AK;Persidsky Y
• 通讯作者: Persidsky Y

let-7 microRNAs: Their Role in Cerebral and Cardiovascular Diseases, Inflammation, Cancer, and Their Regulation.

• DOI: 10.3390/biomedicines9060606
• 发表时间: 2021-05-26
• 期刊: Biomedicines
• 影响因子: 4.7
• 作者: Bernstein DL;Jiang X;Rom S
• 通讯作者: Rom S