Investigating Oatp-Mediated Delivery of Statins to the Brain in Males and Females: Relevance to Neuroprotective Treatment for Ischemic Stroke

研究 Oatp 介导的他汀类药物向男性和女性大脑的递送：与缺血性中风神经保护治疗的相关性

• 批准号: 10682409
• 负责人: Erica Iris Williams
• 金额: $ 4.77万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-22 至 2024-10-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10682409
• 关键词: Acute; Affect; Age; Age Years; Alteplase; Animals; Behavioral; Biological; Blood; Blood - brain barrier anatomy; Brain; Brain Ischemia; Cause of Death; Cerebral Infarction; Cerebral cortex; Clinical; Coenzyme A; Cognitive; Confocal Microscopy; Data; Development; Drug Transport; Edema; Effectiveness; FDA approved; Female; Fibrinolytic Agents; Goals; Gonadal Steroid Hormones; Hemorrhage; Hour; In Situ; Ischemic Stroke; Laboratories; Male Castration; Marketing; Measures; Mechanics; Mediating; Menopause; Methodology; Methods; Middle Cerebral Artery Occlusion; Modeling; Motor; Neurocognitive; Neurocognitive Deficit; Neurologic; Neurological outcome; Neuroprotective Agents; OATP Transporters; Operative Surgical Procedures; Organic Anion Transporters; Outcome; Oxidoreductase; Performance; Perfusion; Pharmaceutical Preparations; Pharmacological Treatment; Pharmacotherapy; Phase III Clinical Trials; Pravastatin; Property; Public Health; Rattus; Recovery of Function; Reperfusion Injury; Research Design; Risk; Role; Severities; Sex Differences; Sprague-Dawley Rats; Stroke; Stroke prevention; Testing; Therapeutic; Time; Tissues; United States; United States National Institutes of Health; Variant; Western Blotting; Woman; Work; age group; atorvastatin; behavior test; cognitive performance; endovascular thrombectomy; experience; experimental study; fexofenadine; improved; indexing; inhibitor; male; men; molecular marker; morris water maze; neurological recovery; neuroprotection; novel; object recognition; pharmacologic; post stroke; preclinical study; protective effect; protein expression; sex; stroke incidence; stroke outcome; stroke patient; stroke therapy; therapeutic effectiveness; therapeutically effective; treatment strategy; uptake

PROJECT SUMMARY Stroke is the fifth leading cause of death in the United States. Approved stroke therapies are limited by narrow treatment windows, the risk of hemorrhagic transformation, and reperfusion injury. Therefore, there is a critical need for neuroprotective drugs that can improve post-stroke neurological performance. Currently, 3-hydroxy-3- methylglutaryl coenzyme A (HMG CoA) reductase inhibitors (i.e., statins) are given to stroke patients due to their proven utility in improving cognitive and motor outcomes. Studies in our laboratory have uncovered a specific biological mechanism that enables statins to be effective drugs for stroke treatment: transport across the blood- brain barrier (BBB) via the endogenous uptake transporter organic anion transporting polypeptide 1a4 (Oatp1a4). We have shown, for the first time, that Oatp-mediated transport is required for atorvastatin to reduce cerebral infarction volume and improve sensorimotor performance at 24 h following transient middle cerebral artery occlusion (tMCAO). We also observed increased atorvastatin uptake in female rats subjected to tMCAO as compared with their age-matched male counterparts; however, it is unknown if these differences in Oatp- mediated transport at the BBB cause variations in atorvastatin’s ability to prevent stroke progression and/or worsening of neurocognitive deficits in the acute/subacute period. Our goals are to assess the role of sex as a biological variable on statin transport in the ischemic brain and to determine how these differences affect statin efficacy as stroke therapeutics. The central hypotheses of this F31 application are i) that functional expression of Oatp1a4 at the BBB is different in males as compared to females following tMCAO; and ii) that statin neuroprotective properties and/or their effects on post-stroke neurological outcomes are influenced by sex-dependent differences in BBB Oatp1a4 activity. Two aims will test these hypotheses: Aim 1: Investigate sex-dependent differences in Oatp1a4-mediated transport of statins at the BBB in stroke. We will perform our studies in male and female SD rats using the tMCAO model. Oatp1a4 localization and protein expression will be assessed using confocal microscopy and western blotting, respectively. Blood-to- brain transport of statins will be measured using in situ brain perfusion, a state-of-the-art methodology to study drug transport at the BBB. Aim 2: Evaluate sex-dependent differences in statin-associated neuroprotection and functional neurological recovery in stroke. In tMCAO operated male and female SD rats, we will use confocal microscopy and western blot analysis to examine molecular biomarkers associated with neuroprotection. We will also assess motor and cognitive performance in tMCAO-animals using robust behavioral tests (i.e., rotarod analysis, Morris Water Maze, Novel Object Recognition test). Overall, this project will provide critical mechanistic data on efficacy of statins as neuroprotective drugs for stroke. Furthermore, this project directly aligns with NIH goals in studying sex as a biological variable in stroke.

项目摘要 中风是美国第五大死亡原因。批准的中风疗法受到狭窄的限制 治疗窗户，出血转化的风险和再灌注损伤。因此，有一个关键 需要可以改善触觉后神经功能表现的神经保护药物。目前，3-羟基-3-- 甲基戊二酰辅酶A（HMG COA）降低了抑制剂（即他汀类药物），因为他们的中风患者因其抑制剂而降低了抑制剂 在改善认知和运动结果方面有证明的实用性。我们实验室的研究发现了特定的 使他汀类药物成为中风治疗的有效药物的生物学机制：跨血液的运输 通过内源摄取转运蛋白有机阴离子运输多肽1A4（OATP1A4）的脑屏障（BBB）。 我们首次表明，Atorvastatin需要OATP介导的运输来减少脑 暂时性大脑伪影后24小时的梗塞体积并改善感觉运动性能 闭塞（TMCAO）。我们还观察到在接受TMCAO AS的雌性大鼠中的阿托伐他汀摄取量增加 与年龄匹配的男性相比；但是，尚不清楚OATP-的这些差异是否存在 BBB的介导运输会导致阿托伐他汀预防中风进展和/或 在急性/亚急性期间神经认知缺陷的恶化。我们的目标是评估性的作用 缺血性大脑中他汀类药物转运的生物变量，并确定这些差异如何影响他汀类药物 作为中风疗法的功效。该F31应用程序的中心假设是i）功能 与TMCAO之后的女性相比，男性在BBB上的OATP1A4表达不同。和II） 他汀类药物神经保护特性和/或它们对中风后神经系统结果的影响是 受BBB OATP1A4活性的性依赖性差异的影响。两个目标将检验以下这些假设： AIM 1：研究OATP1A4介导的他汀类药物在BBB中的性别依赖性差异 中风。我们将使用TMCAO模型对男性和女性SD大鼠进行研究。 OATP1A4本地化 将分别使用共聚焦显微镜和蛋白质印迹评估蛋白质表达。鲜血 他汀类药物的脑运输将使用原位脑灌注来测量，这是一种研究的最先进方法论 BBB的药物运输。 目标2：评估与他汀类药物相关神经保护和功能的性别依赖性差异 中风中的神经恢复。在TMCAO操作的男性和女性SD大鼠中，我们将使用共聚焦显微镜 和蛋白质印迹分析以检查与神经保护相关的分子生物标志物。我们还将评估 使用强大的行为测试（即rotarod分析，莫里斯 水迷宫，新颖的对象识别测试）。 总体而言，该项目将提供有关他汀类药物作为中风的神经保护药有效性的关键机理数据。 此外，该项目直接与NIH目标相吻合，以研究性作为中风的生物变量。