Combination drug treatment to inhibit multiple cell death pathways after TBI

抑制 TBI 后多种细胞死亡途径的联合药物治疗

• 批准号: 7985713
• 负责人: ALAN Ira FADEN
• 金额: $ 48.62万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-11-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7985713
• 关键词: Cell Death; Drug Combinations; Pathway interactions; Cell Death; Drug Combinations; Pathway interactions

DESCRIPTION (provided by applicant): This application entitled "Attenuation of Diverse Cell Death Pathways after Traumatic Brain Injury by Multi- drug Combination Therapy" addresses Broad Challenge Area (15): Translational Science and Specific Challenge Topic: 15-HD-104, Multi-drug Combination Therapy for TBI and Stroke Treatment. The purpose of the proposed studies is to evaluate a novel combination treatment strategy in experimental traumatic brain injury (TBI). Traditional neuroprotective treatment strategies for TBI aim to prevent delayed (secondary) neuronal cell death, generally by inhibiting one proposed cell death mechanism. Yet considerable research indicates that multiple pathways and mechanisms of cell death contribute to tissue loss. In focal TBI, the central injury site is thought to largely reflect necrotic cell death, which primarily occurs within the first 6-8 hours after trauma and is associated with severe bioenergetic compromise. Our recent work, however, indicates that in several rodent TBI models -mouse controlled cortical impact (CCI) or rat lateral fluid percussion (LFP)- cell death in the central core region also includes a substantial component of caspase-independent apoptosis, whereas the better known caspase-dependent cell death is detected in the more peripheral regions where ATP and ADP levels are largely preserved. The caspase-independent programmed cell death (PCD) is due in large part to the activation/translocation of apoptosis-inducing factor (AIF). Such cell death occurs relatively late after injury (24-72h) and can be inhibited by delayed treatment hours after the insult. These data raise serious questions about classical assumptions regarding mechanism of post-traumatic brain injury, suggesting the possibility of newer treatment approaches with extended therapeutic windows and explaining why, if used exclusively, caspase inhibitors may have only a partial protective effect. Ideally, treatment for TBI should attempt to inhibit both caspase-independent and caspase-dependent PCD. Recent studies in ischemia, as well as unpublished work from our laboratory, suggest two intriguing approaches for limiting the three major pathways of cell death after injury. One approach is to up-regulate heat shock protein 70 (HSP70), which binds both Apaf-1 and AIF at distinct sites, thereby neutralizing their pro- apoptotic functions by preventing the formation of the apoptosome (and caspase 3) and attenuating AIF mediated actions. Our preliminary data show that TBI causes up-regulation of HSP70 in many neurons within the injury zone; those neurons expressing HSP 70 show neither caspase-3 activation nor AIF translocation. Up-regulation of HSP70 in cerebral ischemia is strongly protective. The other approach is to inhibit PARP-1. PARP-1, through the release of poly ADP ribose (PAR), is a critical upstream activator AIF release from neuronal mitochondria; it also more recently has been shown to be a critical activator of microglia. Inhibition of PARP-1 after TBI strongly attenuates both caspase dependent and independent forms of PCD, as well as microglial activation, with markedly improved outcome. The advantage of each of these strategies is that their therapeutic window should be very broad, at least 24h. By combining these distinct therapeutic strategies, additive or synergistic protective effects may potentially be achieved. Should our hypotheses be supported, concepts regarding treatment of TBI will be markedly altered and target populations for therapeutic intervention considerably expanded. Combination treatment evaluation is proposed in two pathobiologically different models in different species, with the assumption that potent treatment effects duplicated across models and species makes ultimate clinical translation more likely. Specific hypotheses include: 1) HSP-70 inducers or PARP-1 inhibitors attenuate caspase-independent and caspase-dependent PCD after TBI, reducing long-term neurological dysfunction; 2) Each treatment approach has a long therapeutic window of at least 24h; 3) combined therapy with HSP-70 inducers and PARP-1 inhibitors demonstrate additive and/or synergistic effects in both mouse CCI and rat LFP models. We propose the following specific aims: 1) to compare the efficacy, dose response, and therapeutic window of two structurally distinct HSP-70 inducers with regard to attenuation of post-traumatic neuronal cell death and improved functional recovery after moderate CCI injury in mice; 2) to compare the efficacy, dose response and therapeutic window of two structurally distinct PARP-1 inhibitors with regard to attenuation of post-traumatic neuronal cell death and improved functional recovery after moderate CCI injury in mice; 3) to determine whether combined multi-drug therapy with the best HSP-70 inducer and best PARP-1 inhibitor, at optimal doses, has additive or synergistic effects on cell death, microglial activation, and neurodegenerative conditions up to 3 months post-injury. The Centers for Disease Control and Prevention (CDC) defines traumatic brain injury (TBI) as craniocerebral trauma associated with a decreased level of consciousness, amnesia, other neurologic or neuropsychological abnormalities, skull fracture, intracranial lesions, or death. It has been reported that the combined incidence of fatal and hospitalized TBI among all age groups has a median annual incidence of 101 per 100,000. Approximately 20% of TBIs cause death either immediately or during acute hospital care, with estimated annual rates of mild TBI treated only in outpatient facilities or hospital emergency departments (EDs) in the United States were 392 and 540 visits per 100,000, respectively. A disability prevalence of 37% has been reported for TBI patients followed more than one year after hospitalization; based on this figure, the CDC has estimated that nearly 2% of the entire US population has TBI-related disabilities. Traumatic brain injury (TBI) may occur in as many as 22% of troops deployed in Afghanistan and Iraq. Recent studies in ischemia, as well as unpublished work from our laboratory, suggest two intriguing approaches for limiting the three major pathways of cell death after injury. One approach is to up-regulate heat shock protein 70 (HSP70), which binds both Apaf-1 and AIF at distinct sites, thereby neutralizing their pro-apoptotic functions by preventing the formation of the apoptosome (and caspase 3) and attenuating AIF mediated actions. Our preliminary data show that TBI causes up- regulation of HSP70 in many neurons within the injury zone; those neurons expressing HSP 70 show neither caspase-3 activation nor AIF translocation. Up-regulation of HSP70 in cerebral ischemia is strongly protective. The other approach is to inhibit PARP-1. PARP-1, through the release of poly ADP ribose (PAR), is a critical upstream activator AIF release from neuronal mitochondria; it also more recently has been shown to be a critical activator of microglia. Inhibition of PARP-1 after TBI strongly attenuates both caspase dependent and independent forms of PCD, as well as microglial activation, with markedly improved outcome. The advantage of each of these strategies is that their therapeutic window should be very broad, at least 24h. By combining these distinct therapeutic strategies, additive or synergistic protective effects may potentially be achieved. Should our hypotheses be supported, concepts regarding treatment of TBI will be markedly altered and target populations for therapeutic intervention considerably expanded. Combination treatment evaluation is proposed in two pathobiologically different models in different species, with the assumption that potent treatment effects duplicated across models and species makes ultimate clinical translation more likely.

描述（由申请人提供）： 该应用名为“通过多药结合疗法对创伤性脑损伤后各种细胞死亡途径的衰减”解决了广泛的挑战领域（15）：转化科学和特定挑战主题：15-HD-104，用于TBI和中风治疗的多药结合疗法。拟议研究的目的是评估实验性创伤性脑损伤（TBI）中的新型组合治疗策略。 TBI的传统神经保护治疗策略旨在防止延迟（继发性）神经元细胞死亡，通常通过抑制一种建议的细胞死亡机制。然而，大量研究表明，细胞死亡的多种途径和机制有助于组织损失。在局灶性TBI中，中央损伤部位被认为主要反映了坏死细胞死亡，该死亡主要发生在创伤后的头6-8小时内，并且与严重的生物能妥协有关。 Our recent work, however, indicates that in several rodent TBI models -mouse controlled cortical impact (CCI) or rat lateral fluid percussion (LFP)- cell death in the central core region also includes a substantial component of caspase-independent apoptosis, whereas the better known caspase-dependent cell death is detected in the more peripheral regions where ATP and ADP levels are largely preserved.与caspase无关的编程细胞死亡（PCD）很大程度上归因于诱导凋亡因子的激活/易位（AIF）。这种细胞死亡发生在受伤后（24-72H）后相对较晚，可以通过侮辱后的延迟治疗来抑制。这些数据引发了有关创伤后脑损伤机制的经典假设的严重问题，这表明具有扩展治疗窗口的新治疗方法的可能性，并解释了为什么如果仅使用，caspase抑制剂可能只有部分保护作用。理想情况下，对TBI的治疗应尝试抑制caspase独立和caspase依赖性PCD。 最新的缺血研究以及我们实验室未发表的工作提出了两种有趣的方法来限制受伤后细胞死亡的三个主要途径。一种方法是上调热休克蛋白70（HSP70），该方法在不同的位点结合了APAF-1和AIF，从而通过防止形成凋亡组（和caspase 3）并衰减AIF MEDIADED作用来中和它们的凋亡功能。我们的初步数据表明，TBI在损伤区内许多神经元中引起HSP70的上调。那些表达HSP 70的神经元既不显示caspase-3激活也不显示AIF易位。脑缺血中HSP70的上调具有强烈的保护性。另一种方法是抑制PARP-1。 PARP-1通过释放Poly ADP核糖（PAR）是从神经元线粒体中释放的关键上游激活剂AIF释放。最近，它也被证明是小胶质细胞的关键激活剂。 TBI后对PARP-1的抑制严重减弱了caspase依赖性和独立形式的PCD以及小胶质细胞激活，并明显改善的结果。每种策略的优点是它们的治疗窗口应该非常广泛，至少应该24小时。通过结合这些独特的治疗策略，可以实现添加剂或协同保护作用。如果支持我们的假设，有关TBI治疗的概念将受到明显改变，则针对治疗干预措施的目标大大扩展。在不同物种的两个病理学不同模型中提出了组合治疗评估，假设在模型和物种之间重复的有效治疗效果使最终的临床翻译更有可能。 特定的假设包括：1）HSP-70诱导剂或PARP-1抑制剂减弱了TBI后caspase非依赖性和caspase依赖性PCD，从而减少了长期神经功能障碍； 2）每种治疗方法的治疗窗口至少为24小时； 3）与HSP-70诱导剂和PARP-1抑制剂结合治疗在小鼠CCI和大鼠LFP模型中都表现出添加剂和/或协同作用。 我们提出以下具体目的：1）比较两个结构上不同的HSP-70诱导剂的功效，剂量反应和治疗窗口，以减轻创伤后神经元细胞死亡的衰减，并改善小鼠中等CCI损伤后功能恢复； 2）在比较两种结构上不同的PARP-1抑制剂的功效，剂量反应和治疗窗口，以减轻小鼠后CCI损伤后的创伤后神经元细胞死亡的衰减以及改善功能恢复； 3）确定以最佳剂量的最佳HSP-70诱导剂和最佳的PARP-1抑制剂将多药疗法与最佳剂量相结合的，对细胞死亡，小胶质细胞激活以及伤害后的神经退行性疾病具有添加剂或协同作用。疾病控制与预防中心（CDC）将创伤性脑损伤（TBI）定义为颅骨创伤，与意识水平降低相关，健忘症，其他神经系统或神经心理学异常，颅骨骨折，内部病变或死亡。据报道，所有年龄段的致命性和住院性TBI的综合发生率的中位年发生率为每100,000个101。大约20％的TBI立即或急性医院护理期间死亡，估计仅在美国门诊设施或医院急诊部门（EDS）的年度轻度TBI率分别为每100,000次的392和540次访问。据报道，TBI患者的残疾患病率为37％。基于这一数字，疾病预防控制中心估计，整个美国人口中，近2％的人患有与TBI相关的残疾。在阿富汗和伊拉克部署的部队中，可能有多达22％的部队发生创伤性脑损伤（TBI）。最新的缺血研究以及我们实验室未发表的工作提出了两种有趣的方法来限制受伤后细胞死亡的三个主要途径。一种方法是上调热激蛋白70（HSP70），该方法在不同的位点结合了APAF-1和AIF，从而通过防止形成凋亡组（和caspase 3）并衰减AIF MEDIADID CANCATIO，从而中和它们的促凋亡功能。我们的初步数据表明，在损伤区内许多神经元中，TBI导致HSP70的调节。那些表达HSP 70的神经元既不显示caspase-3激活也不显示AIF易位。脑缺血中HSP70的上调具有强烈的保护性。另一种方法是抑制PARP-1。 PARP-1通过释放Poly ADP核糖（PAR）是从神经元线粒体中释放的关键上游激活剂AIF释放。最近，它也被证明是小胶质细胞的关键激活剂。 TBI后对PARP-1的抑制严重减弱了caspase依赖性和独立形式的PCD以及小胶质细胞激活，并明显改善的结果。每种策略的优点是它们的治疗窗口应该非常广泛，至少应该24小时。通过结合这些独特的治疗策略，可以实现添加剂或协同保护作用。如果支持我们的假设，有关TBI治疗的概念将受到明显改变，则针对治疗干预措施的目标大大扩展。在不同物种的两个病理学不同模型中提出了组合治疗评估，假设在模型和物种之间重复的有效治疗效果使最终的临床翻译更有可能。