The Next Step: Regadenoson mitigates ischemia reperfusion injury and is renal protective in a 48 hour porcine model of resuscitative ECMO

这是一份职业发展奖的申请，旨在为PI提供翻译研究方面的培训。 在他的导师进行的缺血再灌注损伤研究的基础上，PI将使用他的急性 复苏的猪动物模型，并将其延长至48小时以观察终末器官损伤/恢复。这将 进一步将他的职业道路与他的导师分开，并为他提供临床试验的资金。 心脏骤停是世界范围内死亡的主要原因之一， 氧合（ECMO）已成为难治性心脏骤停的治疗选择， 2017年全球约有4,000名患者。当前提案的重点是在我们成功的6- 4小时ECMO猪模型，并将其延长至48小时，以充分评估器官损伤和恢复。我们也 使用FDA批准的A2 AR激动剂（Regadenoson），以最大限度地提高临床可翻译性， 治疗患者。该项目代表了ECMO管理从支持性 作用，积极治疗的根本问题，IRI器官损伤。复苏型ECMO是最极端的 IRI的例子，但这种治疗将适用于所有未来的复苏机械循环支持。 具体目标1A将显示类伽腺苷在48-复苏模型中减弱全身器官损伤。 体外膜肺氧合我们将研究中枢神经系统、心脏、肝脏、肾脏和肠道损伤以及细胞因子的几种标志物。 程度. CD 3+和CD 4 + T细胞、单核细胞和中性粒细胞以及几种损伤相关分子模式 将评估DAMP分子，包括HMGB 1、游离DNA片段和线粒体DNA。目的 图1B显示在用Regadenoson治疗的组中改善的器官功能。心脏功能将 通过Swan-Ganz导管和超声心动图（ECHO）测量的心脏指数以及 正性肌力支持量、IV液体需求量和ECMO流量。将采集尿液排出量， 测量肠通透性，并通过异硫氰酸荧光素（FITC）管饲法测定肠通透性。具体 Aim 2将显示Regadenoson如何通过几种途径减轻器官损伤并改善器官功能 机制包括中性粒细胞活化减弱，细胞凋亡减少，细胞膜 屏障渗透性和肝、肾、肠、心脏和脑中较少的活性氧。这 这一机制的目的将为使用A2 AR激活来减弱IRI提供额外的科学支持。 多器官系统Laubach博士是一名高级研究员，他研究IRI已有二十多年， 将作为该补助金的这一目标的导师。具体目标3将提出一项临床试验，以确定是否 Regadenoson为停搏后患者的多器官系统提供了持久的临床改善 再灌注复苏ECMO。我将使用从我的课程工作和指导中获得的技能， Drs. Lau和Kron在该项目的第四年提交FDA研究性新药申请， 这样我就可以在最后一年提交R 01临床试验资金申请。

This is an application for a career development award to provide the PI with training in translation research. Building on ischemia reperfusion injury research performed by his mentors, the PI will use his acute resuscitative porcine animal model and extend it to 48 hours to look at end organ injury/recovery. This will further separate his career path from his mentors and set him up for funding for a clinical trial. Cardiac arrest is one of the leading causes of death worldwide, and resuscitative extracorporeal membrane oxygenation (ECMO) has emerged as a therapeutic option for refractory cardiac arrest, which was reported in almost 4,000 patients worldwide in 2017. The focus of the current proposal is to build upon our successful 6- hour ECMO porcine model and extend it to 48 hours to fully assess organ injury and recovery. We are also using an FDA-approved A2AR agonist (Regadenoson) to maximize the clinical translatability and to bring this therapy to patients. This project represents a paradigm shift in the management of ECMO from a supportive role to active treatment of the underlying problem, IRI organ injury. Resuscitatve ECMO is the most extreme example of IRI, but this treatment will be applied to all future resuscitative mechanical circulatory support. Specific Aim 1A will show that Regadenoson attenuates systemic organ injury in a 48-model of resuscitative ECMO. We will look at several markers of CNS, cardiac, liver, kidney, and intestinal injury as well as cytokine levels. CD3+ and CD4+ T cells, monocytes and neutrophils, and several damage-associated molecular pattern (DAMP) molecules will be assessed including HMGB1, cell-free DNA fragments and mitochondrial DNA. Aim 1B will show improved organ function in the groups treated with Regadenoson. Cardiac function will be assessed by cardiac index measured by a Swan-Ganz catheter and echocardiographs (ECHO) as well as the amount of inotropic support, IV fluid requirement and ECMO flows. Urine output will be collected and measured, and intestinal permeability will be determined via fluorescein isothiocyanate (FITC) gavage. Specific Aim2 will show how Regadenoson attenuates organ injury and improves organ function through several mechanisms including attenuation of neutrophil activation with reduced apoptosis, decreased membrane barrier permeability and fewer reactive oxygen species in the liver, kidney, intestine, heart, and brain. This mechanistic aim will provide additional scientific support for the use of A2AR activation to attenuate IRI in multiple organ systems. Dr Laubach is a senior researcher who has studied IRI for more than two decades and will serve as a mentor for this aim of the grant. Specific Aim3 will propose a clinical trial to determine if Regadenoson provides a durable clinical improvement in multiple organ systems in patients with post-arrest reperfusion with resuscitative ECMO. I will use the skills obtained from my course work and mentoring from Drs. Lau and Kron to submit a FDA investigational new drug application during the fourth year of this project, so I can submit an R01 proposal for clinical trial funding during the final year.