Targeting Blood-Brain Barrier Transporters to Treat Ischemic Stroke

靶向血脑屏障转运蛋白治疗缺血性中风

基本信息

批准号：
10531891
负责人：
Patrick Thomas Ronaldson
金额：
$ 55.96万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10531891
关键词：
Activin Receptor Age Agonist Alteplase Animal Model Animals Apoptosis Autophagocytosis Biological Markers Blood Blood - brain barrier anatomy Blood Vessels Brain Brain Ischemia Central Nervous System Central Nervous System Agents Clinic Coenzyme A Data Development Dose Drug Delivery Systems Effectiveness Female Goals Grant Hypoxia Incidence Injections Intervention Intravenous Ischemic Stroke Laboratories Measures Mediating Middle Cerebral Artery Occlusion Modeling Motor Necrosis Neurocognitive Neurologic Neurological outcome Neuroprotective Agents OATP Transporters Organic Anion Transporters Oxidative Stress Oxidoreductase Patients Performance Pharmaceutical Preparations Pharmacotherapy Phase III Clinical Trials Pravastatin Process Public Health Publishing Rattus Reperfusion Therapy Rodent Role Signal Transduction Smad Proteins Sprague-Dawley Rats Stress Stroke TGF Beta Signaling Pathway TGF-beta type I receptor Tail Testing Therapeutic Thrombolytic Therapy Time Transforming Growth Factor beta Transforming Growth Factors Translating United States Vascular Endothelium Veins Work activin receptor-like kinase 1 antagonist atorvastatin bone morphogenetic protein 9 brain endothelial cell clinically relevant disability drug discovery endovascular thrombectomy experience experimental study functional outcomes improved in vivo in vivo Model indexing inhibitor intravenous administration male neuroprotection novel novel therapeutics post stroke pre-clinical preclinical study receptor response stroke model stroke outcome stroke patient stroke recovery stroke therapy therapeutic effectiveness thrombolysis treatment strategy uptake

项目摘要

PROJECT SUMMARY Ischemic stroke is a significant public health concern in the United States. Current therapeutic approaches for stroke involve thrombolytic therapy with recombinant tissue plasminogen activator (r-tPA) or endovascular treatments; however, many patients still experience disability. The goal of improving post-stroke outcomes requires novel neuroprotective drugs for stroke treatment. While many such compounds have been identified in preclinical stroke studies, none of these have been successfully translated to the clinic. In contrast, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (i.e., statins) are routinely given to stroke patients due to an inherent ability of these drugs to improve post-stroke functional outcomes. In this grant, we will test the mechanistic hypothesis that neuroprotection from statins results from transport across the blood-brain barrier (BBB) that is mediated by the critical uptake transporter organic anion transporting polypeptide 1a4 (Oatp1a4). This hypothesis will be tested by two aims. Aim 1: To investigate CNS delivery of statins mediated by Oatp1a4 in ischemic stroke. To evaluate the role of Oatp1a4 in CNS drug delivery during stroke, we will investigate Oatp1a4-mediated statin transport at the BBB using the transient middle cerebral artery occlusion (tMCAO) model. In these studies, age-matched male and female rats will be subjected to tMCAO for 90 min (Aim 1A). We will then demonstrate that Oatp1a4- mediated statin delivery improves both biomarkers of neuroprotection and BBB integrity (Aim 1B). We will also perform neurocognitive, sensorimotor, and motor performance studies (i.e., functional neurological tests) in animals administered statins and subjected to tMCAO with reperfusion times of up to 21 days (Aim 1C). In all studies, statins will be administered intravenously either at the time of reperfusion or after 2 h of reperfusion to show that early administration of neuroprotective drugs can improve post-stroke outcomes. Aim 2: Transforming Growth Factor-b (TGF-b) signaling can be targeted to control Oatp1a4- mediated CNS statin delivery in ischemic stroke. In these experiments, we will perform dose-response studies and multiple-dosing experiments in age-matched male and female Sprague-Dawley rats using the TGF- b/ALK1 agonist bone morphogenetic protein (BMP)-9 and the TGF-b/ALK5 antagonist SB431542. We will study the activation of specific Smad proteins that control TGF-b signaling in brain microvascular endothelial cells (Aim 2A). We will also determine the time course of Oatp1a4 expression and activity changes following BMP-9 or SB431542 treatment and their effects on CNS delivery of statins (Aim 2B). Additionally, we will measure indices of neuroprotection, markers of BBB protection, and neurological outcomes in rats subjected to tMCAO, administered BMP-9 or SB431542, and injected intravenously with a statin (Aim 2C). Overall, these studies are clinically relevant because they will demonstrate the effective BBB transport mechanisms are required to confer effectiveness of neuroprotective drugs in stroke.

项目摘要缺血性中风在美国是一个重大的公共卫生问题。当前的治疗性中风的方法涉及重组组织纤溶酶原激活剂（R-TPA）或血管内治疗；但是，许多患者仍然患有残疾。改善后击的目的结果需要新型的神经保护药进行中风治疗。虽然许多这样的化合物已经在临床前中风研究中鉴定出来，这些都没有成功地转化为诊所。相比之下， 3-羟基-3-甲基戊二酰辅酶A（HMG-COA）还原酶抑制剂（即他汀类药物）通常被赋予中风患者由于这些药物的固有能力改善了中风后功能结果。在这笔赠款中，我们将测试汀类药物结果神经保护的机理假设从关键吸收蛋白介导的血脑屏障（BBB）的运输有机阴离子运输多肽1A4（OATP1A4）。该假设将通过两个目标进行检验。目的1：研究由OATP1A4在缺血性中风中介导的汀类药物的CNS递送。评估 OATP1A4在中风期间CNS药物递送中的作用，我们将研究OATP1A4介导的他二介导的他汀类药物的转运使用瞬时脑动脉闭塞（TMCAO）模型的BBB。在这些研究中，年龄匹配雄性和雌性大鼠将接受TMCAO 90分钟（AIM 1A）。然后，我们将证明OATP1A4- 介导的他汀类药物的递送改善了神经保护的生物标志物和BBB完整性（AIM 1B）。我们也会在对神经认知，感觉运动和运动性能研究（即功能性神经系统测试）进行动物服用他汀类药物，并接受TMCAO的再灌注时间长达21天（AIM 1C）。总的来说研究，他汀类药物将在再灌注时静脉注射或再灌注2小时后表明神经保护药的早期给药可以改善触摸后结果。 AIM 2：转化生长因子-B（TGF-B）信号传导可以针对控制OATP1A4- 缺血性中风中的CNS他汀类药物递送。在这些实验中，我们将执行剂量反应使用TGF-的研究和对年龄匹配的男性和女性Sprague-Dawley大鼠进行的研究和多剂量实验 B/ALK1激动品骨形态发生蛋白（BMP）-9和TGF-B/ALK5拮抗剂SB431542。我们将学习在脑微血管内皮细胞中控制TGF-B信号传导的特定SMAD蛋白的激活（AIM 2a）。我们还将在BMP-9或 SB431542治疗及其对他汀类药物中枢神经系统递送的影响（AIM 2B）。此外，我们将测量指数神经保护作用，BBB保护的标志物以及受TMCAO的大鼠的神经系统结局，施用BMP-9或SB431542，并用他汀类药物静脉注射（AIM 2C）。总体而言，这些研究在临床上是相关的，因为它们将证明有效的BBB 需要运输机制来赋予神经保护药在中风中的有效性。