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.

项目摘要心脏骤停（CA）仍然是美国最普遍的残疾和死亡原因之一在过去的一年中，超过550,000个院外和院外CA。只有12％的院外CA患者存活出院和85％的人经历了轻度至严重的神经认知缺陷。缺血 - 重新灌注（I/R）引起的神经元死亡发生在CA后数小时到几天，这可能是其高发病率和死亡率的原因。不幸的是，I/R已证明对大多数治疗方法具有抗药性，一个例外是针对性的温度管理。目前尚无有效的基于药学的干预措施来治疗大脑I/R。识别事实证明，可吸毒的目标很困难。目标来源是一个有望的来源线粒体，因为线粒体功能障碍是再灌注损伤的标志。 I/R诱导线粒体聚（ADP-核糖）聚合酶1（MT-PARP1），加剧ATP部署和细胞死亡。有趣的是，可以在数小时之前观察到MT-PARP1活性和相应的线粒体功能障碍检测经典的核PARP1活性。鉴于氧化受损的DNA是关键触发因素，这是有道理的对于PARP激活和损伤的线粒体是氧化物的关键来源。与pro-DNA修复活动不同核PARP1，pARP1 - / - 动物的麦特纳完整性和线粒体功能增加。所以，我们假设选择性抑制MT-PARP1将通过降低能量来增加神经元存活 CA后的耗竭和改善线粒体功能 - 同时，核PARP1的有益活性。在此提案中，我们旨在检验这一假设。我们已经概念化并综合了一本小说线粒体特异性PARP1抑制剂（MT-Veliparib）。我们将验证MT-的线粒体靶向特异性 Veliparib（AIM 1.1），确定其初步药代动力学和毒性概况（AIM 1.2，1.3），确认其体外功效（AIM 2.1）并验证假设的作用机理（AIM 2.2）。早期实验具有显示MT-Veliparib显着（在统计学上和统计上）显着减少了氧气 - 葡萄糖的剥夺和谷氨酸兴奋性诱导的神经元死亡。随访体内实验将测量MT-的效率 Veliparib在CA的鼠模型中，评估其对生存和功能结果的影响（AIM 3）。这些实验不仅将确定MT-PARP1在I/R中的作用，而且还将证明它们的可行性我们可推广的半ramicidin介导的线粒体靶向策略。最重要的是MT-PARP1 抑制提供了阐明神经元I/R和其他线粒体的有效吸毒靶标目前不存在的疾病。