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.

项目概要 中风是美国第五大死因。批准的中风疗法受到狭窄的限制 治疗窗口、出血转化风险和再灌注损伤。因此，有一个关键的 需要可以改善中风后神经功能的神经保护药物。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）由于其 已证明在改善认知和运动结果方面具有实用性。我们实验室的研究发现了一个特定的 使他汀类药物成为治疗中风的有效药物的生物学机制：跨血液转运 通过内源性摄取转运蛋白有机阴离子转运多肽 1a4 (Oatp1a4) 进入脑屏障 (BBB)。 我们首次证明，Oatp 介导的转运是阿托伐他汀减少脑损伤所必需的。 大脑中动脉短暂性损伤后 24 小时，可减少梗塞体积并改善感觉运动性能 闭塞（tMCAO）。我们还观察到接受 tMCAO 治疗的雌性大鼠阿托伐他汀摄取量增加 与同龄男性相比；然而，尚不清楚 Oatp 中的这些差异是否 BBB 介导的转运会导致阿托伐他汀预防中风进展的能力发生变化和/或 急性/亚急性期神经认知缺陷恶化。我们的目标是评估性别作为 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 作为中风治疗的功效。该 F31 应用的中心假设是 i) 函数 tMCAO 后，男性与女性 BBB 处 Oatp1a4 的表达不同；和 ii) 他汀类药物的神经保护特性和/或其对中风后神经系统结果的影响是 受 BBB Oatp1a4 活性的性别依赖性差异的影响。有两个目标将检验这些假设： 目标 1：研究 Oatp1a4 介导的他汀类药物在 BBB 转运中的性别依赖性差异 中风。我们将使用 tMCAO 模型对雄性和雌性 SD 大鼠进行研究。 Oatp1a4 本地化 和蛋白质表达将分别使用共聚焦显微镜和蛋白质印迹进行评估。血到- 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 BBB 的药物运输。 目标 2：评估他汀类药物相关神经保护和功能的性别依赖性差异 中风的神经功能恢复。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 和蛋白质印迹分析来检查与神经保护相关的分子生物标志物。我们还将评估 使用稳健的行为测试（即旋转分析、Morris 水迷宫、新物体识别测试）。 总体而言，该项目将为他汀类药物作为中风神经保护药物的功效提供关键的机制数据。 此外，该项目与 NIH 研究性别作为中风生物学变量的目标直接一致。