Translational Technologies for Ameliorating Brain Injury

摘要 心脏骤停（CA）对生存具有毁灭性的后果，即使在成功的脑复苏后， 伤害可能相当严重。我们研究的广泛目标是开发转化的治疗技术 用于减轻CA后全脑缺血引起的脑损伤。一个流行的解决方案是治疗性低温 （TH）.虽然TH已被证明可以改善结果，但它并不促进唤醒或减少神经功能。 炎症我们现在提出了一种新的和潜在的翻译传递方法，以促进唤醒， 鼻内递送ORXA。此外，我们还重点研究了内在的生物分布和反 树状聚合物在CA后慢性长期存活中的炎性特性。我们提议 我们希望通过这些实验，找到临床上可转化的解决方案。 这项建议是建立在令人兴奋的初步结果之上的。我们发现了一种方法， 通过Orexin-A（ORXA）的递送的食欲素能途径。我们的初步研究支持这一想法 首次表明，脑室内（ICV）ORXA治疗可减少炎症，此外， 增强性唤起这一想法进一步验证了我们的新的定量脑电图（qEEG）监测 技术.我们已经观察到脑损伤和由于CA脑损伤后神经炎症导致的不良结局。 在我们的初步研究中，我们发现树枝状聚合物的摄取，特别是树枝状聚合物-N-乙酰半胱氨酸（D-乙酰半胱氨酸）， NAC），发生在受伤的大脑区域。我们已经证明，树枝状聚合物作为一种靶向治疗， 通过减轻神经炎症、氧化应激和 兴奋性毒性此外，我们将我们的工作扩展到长期观察，并建立性别特异性模型。 对于拟议的调查，我们将利用广泛研究和验证的啮齿动物CA模型， 复苏，建议在男性和女性受试者中进行急性和慢性实验， 全身灌注监测、电生理（qEEG）监测、综合行为 检查和组织病理学分析。 我们的总体假设是鼻内ORXA将启动脑唤醒效应和早期抗炎作用。 反应，而树枝状聚合物纳米疗法，D-NAC，将减少慢性神经炎症;总之，这些 治疗将改善长期生存。该项目的具体目标是： 目的1：确定鼻内ORXA治疗对早期神经生理恢复的治疗效果， CA昏迷后的认知和行为结果。 目的2：确定鼻内ORXA立即应用的抗炎治疗效果的窗口 复苏后。 目的3：证明使用与N-乙酰基-L-乙酰基-N-乙酰基-L-乙酰基偶联的树枝状聚合物的树枝状聚合物纳米疗法的治疗 半胱氨酸（D-NAC），增加生存，改善神经行为，减少慢性神经炎症， 复苏术 目的4：通过顺序使用CA后神经炎症，实现早期唤醒和神经保护作用 鼻内ORXA和D-NAC用于持续的神经保护，从而改善长期神经功能 结果和CA后生存率。 目前用于改善全球性免疫缺陷综合征后的生存率和认知结果的治疗方案非常有限。 由CA引起的局部缺血。我们的双重方法，鼻内ORXA递送和树枝状聚合物介导的靶向， 满足未满足的促进唤醒和减轻患者CA后神经炎症的需求。此外，本发明还 鼻内给药方法一旦得到验证，就应该能够快速地临床转化。总体而言，我们 这项研究为未来的临床研究奠定了基础，这些研究旨在改善患者术后的预后。 复苏术

Abstract Cardiac arrest (CA) has devastating consequences to survival and, even after successful resuscitation brain injury can be quite severe. The broad goal of our research is to develop translational, therapeutic technologies for mitigating brain injury from global ischemia following CA. One prevailing solution is therapeutic hypothermia (TH). While TH has been shown to improve outcome, it does not promote arousal or reduce neuro- inflammation. We now propose a novel and potentially translational delivery approach to promote arousal by intranasal delivery of ORXA. In addition, we also focus on examining the intrinsic bio-distribution and anti- inflammatory properties of dendrimers in a chronic long-term survival after CA. We propose discovery experiments that, we hope to show, will lead to clinically translatable solutions. This proposal is founded on exciting preliminary results. We have discovered an approach to targeting the orexinergic pathway through the delivery of Orexin-A (ORXA). This idea is supported by our preliminary studies that first showed that intra-cerebral ventricle (ICV) ORXA treatment reduces inflammation, and in addition, rapidly enhances arousal. This idea is further validated by our novel quantitative EEG (qEEG) monitoring technology. We have observed brain injury and poor outcome due to neuro-inflammation post-CA brain injury. In our preliminary studies, we found that uptake of dendrimers, specifically Dendrimer- N-acetyl cysteine (D- NAC), occurs at injured brain regions. We have shown that dendrimers serve as a targeted therapeutic technology for neuro-inflammation by attenuating neuro-inflammation, oxidative stress and excitotoxicity. Further, we extend our work to long term observations and set up gender-specific models. For the proposed investigations, we will utilize extensively researched and validated rodent model of CA and resuscitation, propose both acute and chronic experimentation in male and female subjects and carry out the monitoring of systemic perfusion, electrophysiological (qEEG) monitoring, comprehensive behavioral examination, and histopathological analysis. Our overarching hypothesis is that intranasal ORXA will initiate brain arousal effects and early anti-inflammatory response, while dendrimer nanotherapy, D-NAC, will reduce chronic neuro-inflammation; and together, these therapies will improve long term survival. The specific aims of this project are to: Aim 1: Determine the therapeutic effects of intranasal ORXA treatment on early neurophysiological recovery, cognitive and behavioral outcome following post-CA coma. Aim 2: Determine the window of anti-inflammatory therapeutic effects of intranasal ORXA applied immediately post-resuscitation. Aim 3: Demonstrate that treatment with dendrimer nanotherapy using dendrimer conjugated to N-acetyl-L- cysteine (D-NAC), increases survival, improves neurobehavior and reduces chronic neuro-inflammation, after resuscitation. Aim 4: Achieve early arousal and neuroprotective effect from post-CA neuroinflamation by sequentially using of intranasal ORXA and D-NAC for sustained neuroprotection leading to improved long term neurological outcomes and survival post-CA. There are very limited current therapeutic solutions for improving survival and cognitive outcome after global ischemia resulting from CA. Our dual approaches, intranasal ORXA delivery and dendrimer mediated targeting, serve the unmet needs of promoting arousal, and mitigating post-CA neuro-inflammation for patients. Further, the intranasal delivery approach, once validated, should be amenable to rapid clinical translation. Overall, our research lays the groundwork for future clinical studies directed at improving the patient outcome after resuscitation.