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。获批的卒中治疗受到狭窄 治疗窗、出血性转化的风险和再灌注损伤。因此，存在一个关键的 需要神经保护药物，可以改善中风后的神经功能。目前，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）给予中风患者， 在改善认知和运动结果方面的实用性。我们实验室的研究发现了一种 使他汀类药物成为治疗中风的有效药物的生物学机制：通过血液的运输- 通过内源性摄取转运蛋白有机阴离子转运多肽1a 4（Oatp 1a 4）介导脑屏障（BBB）。 我们第一次发现，阿托伐他汀需要Oatp介导的转运来降低脑内 短暂性大脑中动脉栓塞后24小时梗死体积和感觉运动功能改善 闭塞（tMCAO）。我们还观察到，在接受tMCAO的雌性大鼠中阿托伐他汀摄取增加， 与他们的年龄匹配的男性同行相比，然而，不知道这些差异是否在Oatp。 在BBB介导的转运导致阿托伐他汀预防卒中进展的能力的变化，和/或 急性/亚急性期神经认知缺陷恶化。我们的目标是评估性作为一种 缺血性脑中他汀转运的生物学变量，并确定这些差异如何影响他汀 作为中风治疗剂的功效。该F31应用的中心假设是i）功能性的， tMCAO后，与雌性相比，雄性BBB处Oatp 1a 4的表达不同;和ii） 他汀类药物的神经保护特性和/或其对卒中后神经功能结局的影响， BBB Oatp 1a 4活性的性别依赖性差异的影响。两个目标将检验这些假设： 目的1：研究Oatp 1a 4介导的他汀类药物在BBB中转运的性别依赖性差异 中风我们将使用tMCAO模型在雄性和雌性SD大鼠中进行研究。Oatp 1a 4定位 并分别使用共聚焦显微镜和蛋白质印迹法评估蛋白质表达。血- 他汀类药物的脑转运将使用原位脑灌注进行测量，这是一种最先进的研究方法。 药物在血脑屏障中的转运。 目的2：评估他汀类药物相关神经保护和功能性神经保护的性别依赖性差异。 中风后的神经恢复在tMCAO手术的雄性和雌性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。我们亦会评估 tMCAO-动物中的运动和认知表现（即，旋转杆分析，莫里斯 水迷宫，新物体识别测试）。 总体而言，该项目将提供关于他汀类药物作为脑卒中神经保护药物的疗效的关键机制数据。 此外，该项目直接与NIH研究性别作为中风生物学变量的目标相一致。