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

基本信息

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
项目类别：
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

项目摘要

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治疗的目的是恢复缺血性脑部的灌注。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，迫切需要开发能够“拯救”可挽救的脑组织免受损伤和/或保护血脑屏障完整性的新型治疗策略。在 H/R 期间，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治疗的目标。 H/R。目标 1：研究 H/R 期间 Oatp 介导的 CNS 药物递送。在这个目标中，我们将重点关注 Oatp1a4，即啮齿动物 BBB 上的主要药物转运。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-β) 信号调节 Oatp1a4 表达/活性。 dorsomorphin 和 SB431542（两种 TGF-β 受体抑制剂）对 Oatp1a4 的 BBB 功能表达进行评估，以评估该通路的靶向性，以控制CNS 药物递送（目标 1C）：评估 Mrps 在 BBB 上的药理学靶向是否可以保护 H/R 后的 BBB 完整性，谷胱甘肽 (GSH) 是一种重要的 CNS 抗氧化剂，是 Mrp1、Mrp2 和 Mrp4 的底物。因此，我们将研究体内 H/R 诱导的这些 Mrps 在 BBB 表达/活性的变化（目标 2A），然后我们将检查 GSH 及其变化。由于 GSH 转运系统和酶的改变而导致 BBB 上的氧化形式 GSH 二硫化物 (GSSG)（目标 2B）我们将评估核因子红细胞 2 相关因子 2 (Nrf2) 对 BBB 上的 Mrps 和 GSH 合成/代谢酶的调节。 ) 信号传导（目标 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的情况下进行。 (ROS) 清除剂 TEMPOL（即 4-羟基-2,2,6,6-四甲基哌啶-N-氧基）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。发现通过治疗靶向 BBB 内源性转运蛋白来治疗具有 H/R 成分的疾病的新方法。