Targeting blood-brain barrier transporters to treat hypoxia/reoxygenation stress

靶向血脑屏障转运蛋白治疗缺氧/复氧应激

• 批准号: 9094712
• 负责人: Patrick Thomas Ronaldson
• 金额: $ 31.5万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9094712
• 关键词: ABCC1 gene; Acute; Altitude; Altitude Sickness; Animal Model; Animals; Antioxidants; Apoptosis; Applications Grants; Autophagocytosis; Biological Assay; Blood; Blood - brain barrier anatomy; Blood Preservation; Blood flow; Brain; Brain Diseases; Brain Injuries; Cause of Death; Cells; Cerebral Edema; Coenzyme A; Cysteine; Development; Disease; Disulfides; Drug Delivery Systems; Drug Transport; Endothelium; Enzymes; Esters; Exhibits; Family; Functional disorder; Glutamate-Cysteine Ligase; Glutathione; Glutathione Disulfide; Goals; Grant; Health; Heart Arrest; Human; Hypoxia; In Situ; Ischemic Stroke; Laboratories; Ligase; Mediating; Metabolic; Methionine; Modeling; Molecular Target; Morbidity - disease rate; Necrosis; OATP Transporters; Obstructive Sleep Apnea; Oxidation-Reduction; Oxidative Stress; Oxidoreductase; Oxygen; P-Glycoproteins; Pathway interactions; Perfusion; Pharmaceutical Preparations; Prevention; Property; Protein Isoforms; Public Health; Reactive Oxygen Species; Regulation; Rodent; Rodent Model; Role; Signal Transduction; Stress; Stroke; System; Testing; Therapeutic; Time; Tissues; Transforming Growth Factors; Traumatic Brain Injury; United States; base; brain tissue; clinically relevant; improved; in vivo; indexing; inhibitor/antagonist; mind control; neuroprotection; novel; novel strategies; novel therapeutics; nuclear factor-erythroid 2; oxidative damage; receptor; respiratory; stroke therapy; targeted treatment; tempol; therapeutic target; tool; transcription factor; treatment strategy; uptake

DESCRIPTION (provided by applicant): Hypoxia/reoxygenation (H/R) is a "component" of several brain diseases such as traumatic brain injury, acute respiratory syndrome, obstructive sleep apnea, high altitude cerebral edema, acute mountain sickness, cardiac arrest and ischemic stroke. The objective of current stroke therapy is to restore perfusion to ischemic brain; however, considerable brain cellular damage and BBB dysfunction occurs when blood flow/oxygen supply is re-established. Therefore, there is a critical need for development of novel treatment strategies that can "rescue" salvageable brain tissue from damage and/or protect BBB integrity during H/R. In this grant, we will test the hypothesis that organic anion transporting polypeptides (Oatps) and multidrug resistance proteins (Mrps), two families of endogenous BBB transporters, can be targeted for treatment of H/R. Two specific aims will test this hypothesis. Aim 1: To investigate CNS drug delivery mediated by Oatps during H/R. In this aim, we will focus on Oatp1a4, the primary drug transporting Oatp at the rodent BBB. We will investigate, in vivo, Oatp1a4 mediated transport of 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase inhibitors (i.e., statins) (Aim 1A). We will then correlate changes in Oatp1a4-mediated statin transport with indices of neuroprotective and antioxidant efficacy (Aim 1B). Transforming growth factor (TGF-�) signaling regulates Oatp1a4 expression/activity. Therefore, we will evaluate effects of dorsomorphin and SB431542, two TGF-� receptor inhibitors, on BBB functional expression of Oatp1a4 to evaluate targeting of this pathway for control of CNS drug delivery (Aim 1C). Aim 2: To evaluate if pharmacological targeting of Mrps at the BBB protects BBB integrity following H/R. Glutathione (GSH), a critical CNS antioxidant, is a substrate for Mrp1, Mrp2, and Mrp4. Therefore, we will study in vivo H/R-induced changes in expression/activity of these Mrps at the BBB (Aim 2A). We will then examine changes in GSH and its oxidized form GSH disulfide (GSSG) at the BBB resulting from alterations in GSH transport systems and enzymes (Aim 2B). We will evaluate regulation of Mrps and GSH synthetic/metabolic enzymes at the BBB by nuclear factor erythroid 2-related factor 2 (Nrf2) signaling (Aim 2C). Since Nrf2 signaling is activated by oxidative stress, our studies will be conducted in the presence and absence of the reactive oxygen species (ROS) scavenger TEMPOL (i.e., 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl) to determine if therapeutic targeting of oxidative stress can control BBB Mrp-mediated transport. Our goal in this grant is to facilitate discovery of novel approaches for treating diseases with an H/R component by therapeutic targeting of endogenous transporters at the BBB.

描述（申请人提供）：缺氧/复氧（H/R）是多种脑部疾病的“组成部分”，如创伤性脑损伤、急性呼吸综合征、阻塞性睡眠呼吸暂停、高原脑水肿、急性高山病、心脏骤停和缺血性中风。目前中风治疗的目标是恢复缺血脑部的灌注； 然而，当血流/氧气供应重新建立时，会发生相当大的脑细胞损伤和血脑屏障功能障碍。因此，迫切需要开发新的治疗策略，以“拯救”可挽救的脑组织免受损伤和/或在 H/R 期间保护 BBB 完整性。在这笔资助中，我们将测试以下假设：有机阴离子转运多肽 (Oatps) 和多药耐药蛋白 (Mrps) 这两个内源性 BBB 转运蛋白家族可以作为 H/R 治疗的靶标。有两个具体目标将检验这一假设。目标 1：研究 H/R 期间 Oatps 介导的 CNS 药物递送。在此目标中，我们将重点关注 Oatp1a4，它是啮齿动物 BBB 上运输 Oatp 的主要药物。我们将在体内研究 Oatp1a4 介导的 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）的转运（目标 1A）。然后，我们将 Oatp1a4 介导的他汀类药物转运的变化与神经保护和抗氧化功效指数相关联（目标 1B）。转化生长因子 (TGF-�) 信号调节 Oatp1a4 表达/活性。因此，我们将评估dorsomorphin和SB431542（两种TGF-β受体抑制剂）对Oatp1a4 BBB功能表达的影响，以评估该途径控制CNS药物输送的靶向性（目标1C）。目标 2：评估在 H/R 后以 BBB 为靶标的 Mrps 的药理学是否可以保护 BBB 完整性。谷胱甘肽 (GSH) 是一种重要的中枢神经系统抗氧化剂，是 Mrp1、Mrp2 和 Mrp4 的底物。因此，我们将研究体内 H/R 诱导的这些 Mrps 在 BBB 表达/活性的变化（目标 2A）。然后，我们将检查 BBB 处 GSH 及其氧化形式 GSH 二硫化物 (GSSG) 的变化，这些变化是由 GSH 转运系统和酶的变化引起的（目标 2B）。我们将评估核因子红细胞 2 相关因子 2 (Nrf2) 信号传导对 BBB 处的 Mrps 和 GSH 合成/代谢酶的调节 (Aim 2C)。由于 Nrf2 信号传导是由氧化应激激活的，因此我们的研究将在存在和不存在活性氧 (ROS) 清除剂 TEMPOL（即 4-羟基-2,2,6,6-四甲基哌啶-N-氧基）的情况下进行，以确定氧化应激的治疗靶向是否可以控制 BBB Mrp 介导的转运。我们这笔资助的目标是通过针对 BBB 的内源性转运蛋白进行治疗，促进发现治疗具有 H/R 成分的疾病的新方法。