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: 8758930
• 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/8758930
• 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; 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-ß) signaling regulates Oatp1a4 expression/activity. Therefore, we will evaluate effects of dorsomorphin and SB431542, two TGF-ß 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（由应用提供）：缺氧/重氧（H/R）是多种脑部疾病的“成分”，例如脑部疾病，急性呼吸综合征，阻塞性睡眠呼吸呼吸暂停，高海拔大脑水肿，急性山区疾病，心脏抑制，心脏抑制和缺血性中性疾病。当前中风疗法的目的是恢复缺血性大脑的灌注。 然而，当重新建立血流/氧气供应时，会发生相当大的脑细胞损伤和BBB功能障碍。因此，需要开发新的治疗策略，可以在H/R期间“挽救”可挽救的脑组织免受损害和/或保护BBB完整性。在这笔赠款中，我们将检验以下假设：有机阴离子转运多肽（OATP）和多药耐药蛋白（MRP）是内源性BBB转运蛋白的两个家族，可用于治疗H/R。两个具体的目标将检验这一假设。目的1：研究H/R期间由燕麦介导的CNS药物输送。在此目标中，我们将重点关注OATP1A4，这是啮齿动物BBB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）的转运减少抑制剂（即汀类药物）（AIM 1A）。然后，我们将将OATP1A4介导的他汀类药物转运的变化与神经保护和抗氧化剂效率指数相关联（AIM 1B）。转化生长因子（TGF-ß）信号传导调节OATP1A4表达/活性。因此，我们将评估两种TGF-β受体抑制剂多翼蛋白和SB431542对OATP1A4的BBB功能表达的影响，以评估该途径控制CNS药物递送的途径（AIM 1C）。目标2：评估MRP在BBB上的药物靶向是否在H/R后保护BBB完整性。谷胱甘肽（GSH）是一种临界CNS抗氧化剂，是MRP1，MRP2和MRP4的底物。因此，我们将研究体内H/R诱导的在BBB上这些MRP的表达/活性变化（AIM 2A）。然后，我们将检查GSH及其氧化形式的GSH二硫化物（GSSG）的变化，这是由于GSH传输系统和酶的改变而导致的（AIM 2B）。我们将通过核因子2相关因子2（NRF2）信号（AIM 2C）评估BBB在BBB处MRP和GSH合成/代谢酶的调节。由于NRF2信号传导被氧化物胁迫激活，因此我们的研究将在存在和不存在活性氧（ROS）清除型tempol（即4-羟基-2,6,6,6-四甲基二哌啶-N-氧基）的情况下进行，以确定治疗性抗应激的氧化应激靶标是否可以控制氧化抗性的氧化靶标。我们在这笔赠款中的目标是促进通过在BBB处的内源性转运蛋白的热靶向H/R成分来治疗疾病的新方法。