Targeting glutamate carboxypeptidase in perinatal brain injury

靶向谷氨酸羧肽酶在围产期脑损伤中的作用

• 批准号: 9263554
• 负责人: Sujatha Kannan
• 金额: $ 44.34万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9263554
• 关键词: Acids; Adult; Adverse effects; Affect; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Astrocytes; Autistic Disorder; Binding; Brain; Brain Injuries; Cells; Cerebral Palsy; Chemistry; Child; Chronic; Clinical; Cognitive deficits; Dendrimers; Development; Disease; Dose; Drug Kinetics; Effectiveness; Ensure; Enzymes; Extracellular Space; FOLH1 gene; Female; Glutamates; Health; Health Care Costs; Hour; Hydrolysis; Immigration; Infection; Inflammation; Injury; Intravenous; Knowledge; Learning Disorders; Life; Measures; Mediating; Membrane; Metalloproteases; Microglia; Mission; Modeling; Morphology; Motor; Nanotechnology; Neonatal; Neonatal Brain Injury; Neurodevelopmental Disorder; Neuroglia; Neurons; Neuropeptides; Newborn Infant; Organ; Oryctolagus cuniculus; Pathogenesis; Pathologic; Pathway interactions; Patients; Pediatric Brain Injury; Perinatal; Perinatal Brain Injury; Peripheral; Pharmaceutical Preparations; Pharmacodynamics; Pharmacology; Phenotype; Play; Public Health; Research; Rest; Role; Safety; Sensory; Serum; Sex Characteristics; Site; Slice; Staging; Synapses; Testing; Therapeutic; Time; Transforming Growth Factor beta; Transforming Growth Factor beta Receptors; Translational Research; Translations; Treatment Efficacy; Work; abstracting; base; cognitive function; design; disability; disorder prevention; excitotoxicity; extracellular; glial activation; glutamate carboxypeptidase; improved; in utero; in vivo; inhibitor/antagonist; innovation; insight; male; metabotropic glutamate receptor 3; migration; motor deficit; multidisciplinary; nanomedicine; nanoparticle; nanotherapy; neuroinflammation; neuroprotection; novel; novel therapeutic intervention; postnatal; prevent; protective effect; receptor; repaired; response; targeted delivery; targeted treatment; white matter injury

Targeting glutamate carboxypeptidase in perinatal brain injury Project summary/abstract Neuroinflammation and excitotoxicity, mediated by activated astrocytes and microglia, are major pathophysiological mechanisms implicated in maternal inflammation induced brain injury in the perinatal period, resulting in neurodevelopmental disorders such as cerebral palsy (CP). We hypothesize that targeting both these mechanisms will provide maximum neuroprotection and enable normal development. We propose to achieve this by targeting the enzyme glutamate carboxypeptidase II (GCPII), using the potent, selective GCPII inhibitor 2-(3-mercaptopropyl) pentanedioic acid (2-MPPA), in our established rabbit model of maternal inflammation induced CP. Capitalizing on our novel finding that GCPII is selectively upregulated in activated microglia in newborn rabbits with CP, we will deliver 2-MPPA using dendrimer nanoparticles (D-MPPA), specifically to these cells, thereby improving efficacy and reducing peripheral side effects. Inhibition of GCPII will not only prevent hydrolysis of glutamate from the neuropeptide, but will also increase NAAG levels which can independently exert neuroprotective effects by preventing pre-synaptic glutamate release and by increasing synthesis and release of TGF-β1, which in turn promotes normal microglial function and exerts anti- inflammatory effects. Our preliminary studies demonstrate that: (1) Neonatal rabbits with CP have increased brain glutamate levels, microglial `activation' with restrictive microglial migration, and decreased TGF-β1 in the brain, (2) D-MPPA selectively localizes in activated microglia and astrocytes in newborn rabbits with CP but not in controls (3) D-MPPA is more effective than the drug alone in decreasing extracellular glutamate and in increasing TGF-β1 levels in ex vivo brain slices and in mixed glial cultures exposed to LPS, and in improving short-term motor deficits in rabbit kits with CP. Based on these promising results we hypothesize that, inhibition of GCPII by D-MPPA specifically in activated microglia and astrocytes will decrease inflammation, improve microglial morphology and migration, and decrease neuronal and white matter injury, resulting in improvement in motor and cognitive deficits that persist until adulthood, in newborn rabbits with CP. To test this we will (1) Assess the dose-dependent pharmacokinetics and pharmacodynamics in CP and control rabbit kits. (2) Determine the role of D-MPPA and TGF-β1 in improving microglial morphology and migration in ex vivo brain slices from rabbits with CP and (3) Evaluate the short term and long term efficacy of treatment with intravenous D-MPPA in male and female rabbit kits with CP. The proposed work is innovative because it is the first study that is focused on a nanotechnology based approach to target GCPII specifically in activated microglia following brain injury. It takes advantage of the selective localization of dendrimers at sites of inflammation, to develop therapeutic applications in the postnatal period. This Multi-PI proposal will bring new insights to the role of GCPII in neonatal brain injury and will offer a novel therapeutic approach, bringing complementary expertise in neonatal/pediatric brain injury, GCPII chemistry and pharmacology, and dendrimer nanomedicine.

靶向围产期脑损伤中的谷氨酸羧肽酶 项目摘要/摘要 由活化的星形胶质细胞和小胶质细胞介导的神经炎症和兴奋性毒性是主要的 孕产妇炎症中实施的病理生理机制在围产期诱导的脑损伤 周期，导致神经发育障碍，例如脑瘫（CP）。我们假设目标 这两种机制都将提供最大的神经保护作用并使正常发育。我们建议 通过使用有效的选择性，通过靶向谷氨酸羧肽酶II（GCPII）来实现这一目标 GCPII抑制剂2-（3-甲基丙基）戊二酸（2-MPPA），在我们已建立的母体兔模型中 炎症引起的CP。利用我们的小说发现，即GCPII在激活中有选择地更新 带有CP的新生兔子的小胶质细胞，我们将使用树突聚合物纳米颗粒（D-MPPA）提供2-MPPA， 抑制GCPII对这些细胞的抑制，从而提高效率并降低外周副作用。 不仅会防止神经肽水解谷氨酸的水解，而且还会增加NAAG水平 可以通过防止突触前谷氨酸释放和通过 TGF-β1的合成和释放增加，进而促进正常的小胶质细胞功能并发挥抗 炎症作用。我们的初步研究表明：（1）具有CP的新生儿兔子增加了 脑谷氨酸水平，小胶质细胞的“激活”，具有限制性小胶质细胞迁移，并改善了TGF-β1 大脑，（2）D-MPPA在活化的小胶质细胞和星形胶质细胞中有选择性地定位于具有CP的新生兔 在对照组（3）中，D-MPPA在降低细胞外谷氨酸和中的D-MPPA中比单独的药物更有效 在离体脑切片和暴露于LPS的混合神经胶质培养物中增加TGF-β1水平，并改善 短期电动机在用CP的兔子套件中定义。基于这些承诺结果，我们假设抑制 D-MPPA在活化的小胶质细胞和星形胶质细胞中专门通过D-MPPA进行的GCPII会减少注射，改进 小胶质细胞形态和迁移，并减少神经元和白质损伤，从而改善 在持续到成年的运动和认知缺陷中，在带有CP的新生兔子中。为了测试这一点，我们将（1） 评估CP和对照兔子试剂盒中剂量依赖的药代动力学和药效学。 （2） 确定D-MPPA和TGF-β1在改善离体脑中小胶质形态和迁移中的作用 来自CP的兔子的切片和（3）评估静脉内治疗的短期和长期效率 带有CP的男性和女性兔子套件中的D-MPPA。拟议的工作是创新的，因为这是第一个研究 这重点是基于纳米技术的方法，用于针对激活的小胶质细胞专门针对GCPII 脑损伤后。它利用了在炎症部位的选择性定位，以 在产后开发理论应用。该多PI提案将为您带来新的见解 GCPII在新生儿脑损伤中的作用，将提供一种新型的治疗方法，带来完整性 新生儿/小儿脑损伤，GCPII化学和药理学以及树枝状聚合物纳米医学方面的专业知识。