NANOFORMULATIONS OF REDOX ENZYMES FOR TREATMENT OF ISCHEMIC STROKE

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 这项建议的长期目标是开发自由基清除剂氧化还原酶向大脑的递送系统，以减轻缺血性中风患者的神经炎症并增强神经保护。中风的初步治疗包括解除阻塞和恢复血液流动。当急性期结束时，治疗的重点是恢复脑功能和细胞存活。在这个阶段，将氧化还原酶输送到大脑以减少炎症是非常重要的。然而，血脑屏障(BBB)严重限制了治疗性多肽向大脑的输送，是成功治疗许多毁灭性的中枢神经系统(CNS)疾病的主要障碍。为了改善治疗性多肽向大脑的转运，保持酶的活性，降低免疫原性，治疗性氧化还原酶将与相反电荷的合成聚电解质交联，形成稳定的多离子络合物胶束，即纳米酶。我们假设，1)酶纳米容器在生理条件下将是稳定的，2)将增强酶对体外血脑屏障的通透性，3)将增加体内循环时间和/或对血脑屏障的通透性，以及4)将改善体外脑缺氧和体内中风模型的生物活性。其具体目标是1)组装氧化还原纳米酶(超氧化物歧化酶、过氧化氢酶)并定制其组成，以增加稳定性、循环时间和/或跨血脑屏障的通透性；2)确定由SA1合成的纳米酶在体外和体内跨脑微血管内皮细胞(BMVEC)的渗透性和转运机制；3)确定在SA2中筛选出的最有希望的纳米酶在体外脑缺氧和体内中风模型中是否可以减轻神经炎症和提供神经保护。预计这些研究将为跨血脑屏障的再生治疗提供一个新的平台。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The long-term objective of this proposal is to develop delivery system of free-radical scavengers, RedOx enzymes, to the brain to attenuate neuroinflammation and increase neuroprotection in patients with ischemic stroke. Initial treatment for stroke involves removing the blockage and restoring blood flow. When acute phase is over, the treatment focused on restoration of brain function and cell survival. At this stage, delivery of RedOx enzymes to the brain to decrease inflammation is of great importance. However, the blood brain barrier (BBB) severely limits the delivery of therapeutic polypeptides to the brain and is a major obstacle to the successful treatment of many devastating central nervous system (CNS) diseases. To improve the therapeutic polypeptide transport to the brain, preserve enzyme activity, and reduce immunogenecity, the therapeutic RedOx enzymes will be cross-linked with a synthetic polyelectrolyte of opposite charge to form a stable polyion complex micelle, "nanozyme". We hypothesize that 1) enzyme incorporated nanocontainers will be stable at physiological conditions, 2) will enhance permeability of the enzyme across the in vitro BBB, 3) will increase circulation time and/or permeability across the BBB in vivo, and 4) will improve biological activity in in vitro brain hypoxia and in vivo stroke models. Specific Aims are 1) to assemble RedOx nanozymes (SOD, catalase) and tailor the composition for increased stability, circulation time and/or permeability across the BBB, 2) to determine permeability and transport mechanisms of the nanozymes synthesized in SA1 across the brain microvascular endothelial cells (BMVEC) in vitro and in vivo, 3) to determine whether the most promising nanozymes selected in SA2 can attenuate neuroinflammation and provide neuroprotection in an in vitro brain hypoxia and in vivo stroke models. It is anticipated that these studies will provide a novel platform for the delivery of therapeutic proteins across the BBB for regenerative therapy.