Targeting mitochondrial PARP1 in neuronal ischemia-reperfusion injury

靶向线粒体 PARP1 在神经元缺血再灌注损伤中的作用

基本信息

批准号：
9917829
负责人：
Andrew Michael Lamade
金额：
$ 5.05万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-04-30 至 2023-04-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9917829
关键词：
ANXA5 gene Acute Animal Model Animals Bioenergetics Blood - brain barrier anatomy Blood flow Brain Brain Ischemia Brain region C57BL/6 Mouse Cause of Death Cell Culture Techniques Cell Death Cell Nucleus Cell division Cell model Cells Cessation of life Clinical Data Critical Care DNA DNA Damage DNA Repair DNA Repair Enzymes Data Detection Drug Kinetics Electron Transport Energy Metabolism Evaluation Fright Glucose Glutamates Heart Arrest Histologic Hospitals Hour Image In Vitro Induced Heart Arrest Injury Intervention Ischemia Learning Link Mass Spectrum Analysis Measures Mediating Medicine Mitochondria Mitochondrial DNA Mitochondrial Diseases Mitochondrial Proteins Modeling Morbidity - disease rate Neocortex Nervous System Physiology Neurocognitive Deficit Neurons Nuclear Oxidants Oxygen Pathology Patients Pharmaceutical Preparations Pharmacology Physicians Plasma Poly(ADP-ribose) Polymerases Polymerase Process Property Proteins Renal function Reperfusion Injury Reperfusion Therapy Research Resistance Role Scientist Source Specificity Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Temperature Testing Therapeutic Toxic effect Training United States Withdrawal apoptosis inducing factor base career circulating biomarkers clinically relevant conditioned fear deprivation design disability drug discovery druggable target endonuclease G experience experimental study fluoro jade follow-up functional outcomes improved in vivo inhibitor/antagonist injured innovation life-sustaining therapy liver function member mitochondrial dysfunction morris water maze mortality mouse model natural hypothermia neuron loss neuronal survival novel novel therapeutics out-of-hospital cardiac arrest oxidative DNA damage pharmacokinetics and pharmacodynamics preservation prevent programs release factor repaired response societal costs stroke model survival outcome therapeutic target

项目摘要

PROJECT SUMMARY Cardiac arrest (CA) remains one of the most prevalent causes of disability and death in the United States with more than 550,000 in- and out-of-hospital CA in the past year. Only 12% of out-of-hospital CA patients survive to discharge and 85% experience mild to severe neurocognitive deficits. Ischemia-reperfusion (I/R)-induced neuronal death occurs hours to days post-CA and is the likely cause of its high morbidity and mortality. Unfortunately, I/R has proven resistant to most therapeutic approaches, an exception is targeted temperature management. There are currently no effective pharmacologic-based interventions to treat brain I/R. Identifying druggable targets for intervention has proven difficult. One ever promising source of targets is the mitochondria, as mitochondrial dysfunction is a hallmark of reperfusion injury. I/R induces hyperactivation of mitochondrial poly(ADP-ribose)polymerase 1 (mt-PARP1), exacerbating ATP depletion and cell death. Interestingly, mt-PARP1 activity and corresponding mitochondrial dysfunction can be observed hours before detection of classic nuclear PARP1 activity. This makes sense given oxidatively damaged DNA is a key trigger for PARP activation and injured mitochondria are a key source of oxidants. Unlike the pro-DNA repair activity of nuclear PARP1, PARP1-/- animals have increased mtDNA integrity and mitochondrial function. Therefore, we hypothesize that selective inhibition of mt-PARP1 will increase neuronal survival by reducing energetic depletion and improving mitochondrial function post-CA – all while sparing beneficial activity of nuclear PARP1. In this proposal, we aim to test this hypothesis. We have conceptualized and synthesized a novel mitochondria-specific PARP1 inhibitor (mt-veliparib). We will verify the mitochondrial targeting specificity of mt- veliparib (Aim 1.1), determine its preliminary pharmacokinetic and toxicity profile (Aim 1.2, 1.3), confirm its efficacy in vitro (Aim 2.1) and verify the hypothesized mechanisms of action (Aim 2.2). Early experiments have shown mt-veliparib significantly (both in effect size and statistically) reduces oxygen-glucose deprivation and glutamate excitotoxity-induced neuronal death. Follow-up in vivo experiments will measure the efficacy of mt- veliparib in a murine model of CA, assessing its effects on survival and functional outcome (Aim 3). These experiments will not only determine the role of mt-PARP1 in I/R, but they will also demonstrate the feasibility of our generalizable hemigramicidin-mediated mitochondria targeting strategy. Most importantly, mt-PARP1 inhibition offers a chance to elucidate an effective druggable target for neuronal I/R and other mitochondrial disorders where none currently exists.

项目概要心脏骤停（CA）仍然是美国最常见的残疾和死亡原因之一去年有超过 550,000 名院内和院外 CA 患者只有 12% 存活。出院，85% 经历轻度至重度缺血再灌注 (I/R) 引起的神经认知缺陷。神经元死亡发生在 CA 后数小时至数天，可能是其高发病率和死亡率的原因。不幸的是，I/R 已被证明对大多数治疗方法具有抵抗力，目标温度是一个例外目前尚无有效的基于药物的干预措施来治疗脑 I/R。事实证明，可用于干预的药物靶标是困难的。线粒体，因为线粒体功能障碍是再灌注损伤的一个标志。线粒体聚（ADP-核糖）聚合酶 1 (mt-PARP1)，加剧 ATP 耗竭和细胞死亡。提示，mt-PARP1 活性和相应的线粒体功能障碍可以在数小时前观察到。检测经典的核 PARP1 活性是有道理的，因为氧化损伤的 DNA 是一个关键的触发因素。与 DNA 修复活性不同，PARP 激活和受损线粒体是氧化剂的关键来源。核 PARP1、PARP1-/- 动物的 mtDNA 完整性和线粒体功能有所增加。我们加强了选择性抑制 mt-PARP1 将通过减少能量来增加神经元存活率 CA 后消耗并改善线粒体功能，同时保留核 PARP1 的有益活性。在这个提案中，我们旨在测试这个假设，我们已经概念化并综合了一个小说。线粒体特异性 PARP1 抑制剂 (mt-veliparib) 我们将验证 mt- 的线粒体靶向特异性。 veliparib（目标 1.1），确定其初步药代动力学和毒性特征（目标 1.2、1.3），确认其体外功效（目标 2.1）并验证已开发的作用机制（目标 2.2）。显示 mt-veliparib 显着（无论是在效应大小还是在统计上）减少氧葡萄糖剥夺和后续体内实验将测量谷氨酸兴奋性毒性诱导的神经元死亡。在 CA 小鼠模型中使用 veliparib，评估其对生存和功能结果的影响（目标 3）。实验不仅将确定 mt-PARP1 在 I/R 中的作用，还将证明以下方法的可行性：我们的通用半短杆菌肽介导的线粒体靶向策略最重要的是 mt-PARP1。抑制为阐明神经元 I/R 和其他线粒体的有效药物靶标提供了机会目前不存在的疾病。