Translational Technologies for Ameliorating Brain Injury

摘要 心脏骤停(CA)对生存和脑复苏具有破坏性后果，即使在成功复苏后 伤势可能会相当严重。我们研究的广泛目标是开发可翻译的治疗技术 用于减轻CA后全脑缺血所致的脑损伤。一种流行的解决方案是治疗性低温。 (Th)。虽然TH已被证明可以改善预后，但它并不能促进觉醒或减少神经功能。 发炎。我们现在提出了一种新的潜在的翻译交付方法来促进唤醒 ORXA鼻腔给药。此外，我们还重点考察了其内在的生物分布和抗 树枝状大分子在CA术后慢性长期存活中的炎症特性。我们建议发现者 我们希望展示的实验将导致临床上可翻译的解决方案。 这项建议是建立在令人振奋的初步结果之上的。我们发现了一种将目标对准 食欲素-A(Orxine-A，ORXA)的递送途径。这个想法得到了我们的初步研究的支持 这首先表明，脑室内(ICV)ORXA治疗可以减少炎症，此外，还可以迅速 提高唤醒能力。我们的新型定量脑电(QEEG)监测进一步验证了这一思想 技术我们观察到CA脑损伤后由于神经炎症造成的脑损伤和不良预后。 在我们的初步研究中，我们发现树状大分子，特别是树状大分子-N-乙酰半胱氨酸(D- NAC)，发生在受损的大脑区域。我们已经证明树枝状大分子具有靶向治疗作用。 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，并建立了针对性别的模型。 在拟议的调查中，我们将利用经过广泛研究和验证的CA和 复苏，建议在男性和女性受试者中进行急性和慢性实验，并进行 全身血流监测、电生理(QEEG)监测、综合行为监测 检查和组织病理学分析。 我们的主要假设是，鼻腔内的ORXA将启动大脑唤醒效应和早期抗炎 反应，而树枝状大分子纳米疗法，D-NAC，将减少慢性神经炎症；总的来说，这些 治疗将提高长期存活率。该项目的具体目标是： 目的1：观察鼻腔给药对早期神经生理学恢复的影响。 CA昏迷后的认知和行为结局。 目的2：确定鼻腔即刻应用ORXA的抗炎疗效窗口 复苏后。 目的3：论证N-乙酰-L偶联的树枝状大分子纳米治疗 半胱氨酸(D-NAC)，增加存活率，改善神经行为，减少慢性神经炎症，在 复苏。 目的4：在CA术后神经炎性反应的早期唤醒和神经保护作用 鼻腔应用ORXA和D-NAC进行持续神经保护可改善长期神经功能 CA术后的预后和存活率。 目前用来改善存活率和认知结果的治疗方案非常有限 CA引起的缺血。我们的两种方法，鼻腔给药和树枝状大分子介导的靶向， 服务于促进唤醒和减轻CA术后神经炎症的未得到满足的需求。此外， 鼻腔给药方法一旦得到验证，应适用于快速临床翻译。总的来说，我们的 研究为未来的临床研究奠定了基础，旨在改善术后患者的预后 复苏。

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.