Novel Peptides for Resuscitation

心脏骤停（SCA）是美国的主要死亡原因。它影响了超过50万人 总生存率为7%。与其他主要死因不同的是， 以提高SCA的存活率。在心肺复苏（CPR）过程中， 临床前研究对SCA损伤具有高度保护作用，并且似乎是由增强的Akt信号传导介导的。 然而，CPR冷却难以在临床上物理实施。开发新的心肺复苏药物， 不需要物理冷却的冷却保护可以是非常有效和平移的。提出 一项工作使用了两种新的细胞渗透性肽，设计用于抑制PHLPP磷酸酶（TAT-PHLPP 9 c）， 分别激活PDK 1（TAT-PIF）。它们在不到5分钟内到达心脏和大脑等重要器官 当静脉内给药于小鼠时，在延长12小时后协同改善4小时SCA存活率。 心搏停止分钟。猪的初步研究表明，静脉注射达特- CPR期间PHLPP 9 c快速改善ROSC后的心功能恢复，并显著改善24 心室除颤（VF）5分钟后神经系统完整存活1小时。我们假设心肺复苏术 TAT-PHLPP 9 c和TAT-PIF的施用协同诱导了细胞的快速和最大活化， Akt随后增强PDH活性，减少葡萄糖向山梨糖醇的分流，并增强葡萄糖代谢。 利用导致能量的早期补充，减少渗透损伤和牛磺酸的释放 和谷氨酸进入血液，并改善心脏功能和神经系统完好的生存。这项建议 将采取系统的努力来测试一种新的治疗策略及其作用机制， 在心脏骤停后CPR期间给予细胞可渗透肽（TAT-PHLPP 9 c和TAT-PIF）， 小鼠和猪，并进一步验证生物标志物和视神经鞘的非侵入性测量 直径（ONSD）来预测人类SCA结果，如以下三个目标所示。 目标1。确定TAT-PHLPP 9 c（一种新型的PHLPP磷酸酶生物抑制剂）和TAT-PIF（一种 PDK 1的新型生物激活剂，在延长的停滞（12分钟）小鼠SCA模型中改善SCA存活。 目标2.确定这些新的肽是否改善猪SCA生存时，静脉注射给药， CPR，并将这些肽与SCA的主动冷却和ECMO疗法的疗效进行比较。 目标3.用超声波检测渗透压和代谢恢复的生物标志物（牛磺酸和谷氨酸） 测量渗透压相关ONSD以预测SCA结果。 猪和人类在解剖学和生理学上有许多相似之处，因此猪已被广泛应用于 被认为是测试治疗的高度转化模型。如果这两种肽能起到降温的作用， 在SCA之后的死猪中，它们很有可能在人体中起冷却作用，而不需要 这将是SCA复苏的一个重大进步。

Sudden cardiac arrest (SCA) is a leading cause of death in the United States. It affects over 500,000 people annually with an overall survival rate of 7%. Unlike other leading causes of death, no pharmacological drugs exist to improve SCA survival. Cooling a few degrees Celsius during cardiopulmonary resuscitation (CPR) in pre-clinical studies is highly protective against SCA injury and appears mediated by enhanced Akt signaling. However, CPR cooling is difficult to physically implement clinically. Development of new CPR drugs that mimic cooling protection without the need for physical cooling could be highly effective and translational. Proposed work uses two novel cell-permeable peptides designed to inhibit PHLPP phosphatase (TAT-PHLPP9c) and to activate PDK1 (TAT-PIF) respectively. They reach critical organs such as heart and brain in less than 5 min when administering to the mouse intravenously and synergistically improve 4 h SCA survival after prolonged 12 min asystole cardiac arrest. Preliminary work in swine demonstrates that intravenous administration of TAT- PHLPP9c during CPR rapidly improves recovery of cardiac function after ROSC and significantly improves 24 hour neurologically intact survival after 5 min ventricular defibrillation (VF). We hypothesize that CPR administration of TAT-PHLPP9c and TAT-PIF collaboratively induce a rapid and maximal activation of Akt, subsequently enhances PDH activity, reduces glucose shunting to sorbitol, and enhances glucose utilization leading to an early replenishment of energy, diminished osmotic injury and release of taurine and glutamate into blood, and improved cardiac function and neurologically intact survival. This proposal will take systematic efforts to test a novel therapeutic strategy and their mechanisms of action by intravenously administration of cell-permeable peptides (TAT-PHLPP9c and TAT-PIF) during CPR following cardiac arrest in mouse, and swine, and further validate biomarkers and a non-invasive measurement of optic nerve sheath diameter (ONSD) to predict SCA outcome in human as illustrated in the following three aims. Aim 1. Determine whether TAT-PHLPP9c, a novel biological inhibitor of PHLPP phosphatase, and TAT-PIF, a novel biological activator of PDK1, improves SCA survival in a prolonged arrest (12 min) mouse SCA model. Aim 2. Determine whether these novel peptides improve swine SCA survival when given intravenously during CPR and compare the efficacy of these peptides to active cooling and ECMO therapies for SCA. Aim 3. Test biomarkers (taurine and glutamate) of osmotic stress and metabolic recovery with ultrasound measurement of osmotic stress-related ONSD to predict SCA outcome. Swine and human have a number of similarities in anatomy and physiology, therefore swine has been widely accepted as a highly translational model in testing therapies. If these two peptides can work as cooling and bring dead swine back after SCA, they have a high possibility to work in the human as cooling without a need to physically cool the patients which would be a major advance in SCA resuscitation.