Therapeutic Targeting of the Class IB PI3-Kinase Gamma for Treatment of Acute Ischemic Stroke

IB 类 PI3 激酶 Gamma 的治疗靶向治疗急性缺血性中风

• 批准号: 9344701
• 负责人: Guohong Li
• 金额: $ 33.97万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9344701
• 关键词: 1-Phosphatidylinositol 3-Kinase; Acute; Adverse effects; Alteplase; Animals; Blood - brain barrier anatomy; Brain; Brain Infarction; Brain Injuries; Brain hemorrhage; Cause of Death; Cerebral Ischemia; Cerebral hemisphere hemorrhage; Cerebrum; Clinical; Data; Deposition; Endothelium; FDA approved; Fibrin; Gelatinase B; Genetic; Hemorrhage; Hour; Immune response; Infarction; Inflammation; Inhibition of NF-KB activation; Intervention; Investigation; Ischemia; Ischemic Stroke; Laboratories; Lesion; Measurement; Mediating; Microglia; Middle Cerebral Artery Occlusion; Modeling; Molecular; Mus; NF-kappa B; Neurologic Deficit; Neurological outcome; Outcome; Phosphatidylinositols; Phosphotransferases; Plasminogen Activator Inhibitor 1; Protein Isoforms; Proteins; Publishing; Rattus; Reperfusion Therapy; Reporting; Risk; Role; Signal Transduction; Source; Stroke; Stromelysin 1; Testing; Therapeutic; Thromboplastin; Thrombosis; Thrombus; Time; Tissues; United States; Work; acute stroke; animal mortality; attenuation; behavior test; brain endothelial cell; brain tissue; cell type; cerebrovascular; density; disability; dosage; experimental study; improved; inhibitor/antagonist; insight; neurovascular injury; neutrophil; novel; novel therapeutics; public health relevance; small molecule; stroke therapy; therapeutic target

DESCRIPTION (provided by applicant): Therapeutic targeting of the class IB PI3-kinase gamma for treatment of acute ischemic stroke. Recombinant tPA remains the only FDA approved and the most beneficial proven intervention for treatment of ischemic stroke. However, clinical use of tPA is limited due to its narrow therapeutic window and increased risk of hemorrhagic transformation (HT). In this proposal, we hypothesize that therapeutic inhibition of PI3Kg with AS605240, alone or in combination with tPA, protects against the ischemia- and tPA-induced brain injury and HT and increases therapeutic window for tPA treatment of ischemic stroke. The molecular basis for our idea is that genetic deletion of PI3Kg not only reduces stroke-induced BBB disruption and tissue infarction but also almost completely blocked cerebral hemorrhage induced by tPA administered at 6 hours after stroke. Our data are promising and suggest a critical role of PI3Kg in tPA-induced HT and brain injury after ischemic stroke. Further, our pilot data show that inhibition of PI3Kg with AS605240 significantly protects against brain injury and reduces fibrin deposition/thrombosis in the ischemic brain after embolic stroke in rats. These exciting preliminary data suggest that PI3Kg may represent a promising therapeutic target for treating acute ischemic stroke. In this application, we propose to develop a new stroke therapy by therapeutic targeting of PI3Kg with AS605240 for treatment of acute ischemic stroke and to investigate molecular mechanisms underlying the AS605240-mediated beneficial effects. We now propose three aims to investigate this new stroke therapy. In Aim 1, we will determine the effects of AS605240, alone and in combination with tPA, treated at 6 hrs after focal embolic stroke in rats. We will identify the optimal dosage and the extended therapeutic time window for AS605240 alone and in combination with tPA for treatment of acute stroke. Brain infarction, neurological deficits, brain hemorrhage, and animal mortality will be examined 72h after stroke. In Aim 2, we will define the mechanisms that underlie the beneficial effects of AS605240 alone or in combination with tPA in the treatment of acute stroke. BBB damage will be assessed by the spatial and temporal analysis of key BBB proteins and potential correlation with changes in activation/expression of NF-κB, CD147, MMP-9 and/or MMP-3 on cerebral vessels. Thrombus formation in the ischemic brain after stroke will be assessed by analyzing fibrin deposition within and around the cerebral vessel wall and the expression of prothrombotic factors (PAI-1, tissue factor-TF) in the ischemic brain. In Aim 3, we will assess long term outcomes of AS605240 alone and in combination with tPA treated at 6 hrs after embolic stroke. Animals will be followed over 5 weeks with a battery of behavioral tests. At the end, brains are removed for measurements of lesion size, fibrin deposition/thrombosis, cerebral vascular density/potency, and their correlation with the expression of proangiogenic factors (VEGF, Ang-1) and prothrombotic factors (PAI-1, tissue factor-TF). If successful, these proposed experiments should provide new insights of how the class IB PI3Kg signaling contributes to the ischemia- and tPA-induced HT and brain injury and may ultimately yield new therapies to treat acute ischemic stroke and to minimize the adverse effect of tPA on ischemic neurovascular damage, leading to improved long term outcomes after ischemic stroke.

描述（由申请人提供）：IB类PI3激酶γ的治疗靶向用于治疗急性缺血性中风。重组 tPA 仍然是 FDA 唯一批准的、经证实的治疗缺血性中风最有益的干预措施。然而，tPA 的临床使用由于其治疗窗狭窄和出血性转化 (HT) 风险增加而受到限制。在该提案中，我们假设单独使用 AS605240 或与 tPA 联合使用 AS605240 对 PI3Kg 进行治疗性抑制，可防止缺血性和 tPA 诱导的脑损伤和 HT，并增加 tPA 治疗缺血性中风的治疗窗口。我们的想法的分子基础是，PI3Kg 的基因缺失不仅可以减少中风引起的血脑屏障破坏和组织梗塞，而且几乎可以完全阻止中风后 6 小时施用 tPA 引起的脑出血。我们的数据很有希望，表明 PI3Kg 在 tPA 诱导的 HT 和缺血性中风后脑损伤中发挥着关键作用。此外，我们的试验数据表明，用 AS605240 抑制 PI3Kg 可显着防止大鼠栓塞性中风后脑损伤并减少缺血性脑中的纤维蛋白沉积/血栓形成。这些令人兴奋的初步数据表明，PI3Kg 可能代表治疗急性缺血性中风的有希望的治疗靶点。在此应用中，我们建议开发一个 通过使用 AS605240 治疗性靶向 PI3Kg 来治疗急性缺血性中风，并研究 AS605240 介导的有益作用背后的分子机制。我们现在提出三个目标来研究这种新的中风疗法。在目标 1 中，我们将确定 AS605240 单独使用以及与 tPA 联合使用在大鼠局灶性栓塞性中风后 6 小时治疗的效果。我们将确定 AS605240 单独使用以及与 tPA 联合治疗急性卒中的最佳剂量和延长的治疗时间窗。中风后72小时检查脑梗塞、神经功能缺损、脑出血和动物死亡率。在目标 2 中，我们将定义 AS605240 单独或与 tPA 联合治疗急性中风的有益作用的机制。通过对关键 BBB 蛋白的空间和时间分析以及与脑血管上 NF-κB、CD147、MMP-9 和/或 MMP-3 激活/表达变化的潜在相关性来评估 BBB 损伤。通过分析脑血管壁内和周围的纤维蛋白沉积以及缺血脑中促血栓因子（PAI-1、组织因子-TF）的表达来评估中风后缺血脑中的血栓形成。在目标 3 中，我们将评估单独使用 AS605240 以及在栓塞性中风后 6 小时联合使用 tPA 治疗的长期结果。将通过一系列行为测试对动物进行为期 5 周的跟踪。最后，取出大脑以测量病变大小、纤维蛋白沉积/血栓形成、脑血管密度/效力及其与促血管生成因子（VEGF、Ang-1）和促血栓因子（PAI-1、组织因子-TF）表达的相关性。如果成功，这些拟议的实验应该为 IB 类 PI3Kg 信号传导如何导致缺血和 tPA 诱导的 HT 和脑损伤提供新的见解，并可能最终产生治疗急性缺血性中风的新疗法，并最大限度地减少 tPA 对缺血性神经血管损伤的不利影响，从而改善缺血性中风后的长期结果。