Hypothermia for Cardiac Arrest: Optimizing Akt-Nitric Oxide Synthase Signaling

描述(由申请人提供)：这份提案是应聘者在急诊医学和生物医学工程方面背景的自然延伸。他对翻译科学的独特背景和热情已经为动物复苏研究做出了贡献。这项创新的工作模拟了我们患者心脏骤停后再灌注损伤的早期，即尽管初始恢复自主循环(ROSC)，但心血管衰竭。该培训计划将通过正式培训申请者学习研究一氧化氮信号和解偶联的最佳分子生物学方法来加强已经完成的小鼠建模。申请者将受益于紧急复苏中心内的多学科指导，建立在心脏氧化剂损伤和适应以及自由基生物学的专业知识基础上。发起人Vanden Hoek博士和McNally博士都是经验丰富的内科科学家。他们也是氧化剂介导的心肌细胞和小鼠模型心脏收缩功能障碍和死亡的合作者。这个职业发展项目建立在申请者的优势、培训计划以及机构支持和指导的基础上。我们假设，心脏骤停后的保护性低温增加了Akt相关的结构性一氧化氮合酶(CNOS)NO信号，同时减弱了CNOS解偶联和破坏性活性氧(ROS)的产生。具体地说，在心脏骤停的小鼠模型中，我们将：(目标1)基于心脏Akt和cNOS活性优化保护性低温的目标温度；(目标2)测试NOS1/NOS3在心脏骤停后损伤、NO和ROS产生以及使用选择性抑制剂和基因敲除策略的低温保护中的相对重要性；以及(目标3)。测试是否可以通过一氧化氮合酶底物/辅因子补充策略复制低温保护。芝加哥大学医学部和急诊医学部有使用K08医生-科学家培训计划进行成功指导的记录。拟议中的培训将为Beiser博士提供更多技能，使其成为该领域领先的独立内科医生兼科学家。从普通人的角度来看，心脏骤停(当心脏停止跳动时)与电视或电影中的描述不同--大多数最初复苏的人都不会活下来。这项工作将有助于理解身体降温几度实际上是如何增加一种帮助身体自我康复的氧化应激的。

DESCRIPTION (provided by applicant): This proposal is a natural extension of the candidate's background in Emergency Medicine and Biomedical Engineering. His unique background and passion for translational science has already contributed to animal resuscitation research. This innovative work models early post-cardiac arrest reperfusion injury in our patients, i.e. cardiovascular collapse despite initial return of spontaneous circulation (ROSC). The training program will enhance the mouse modeling already done by formally training the applicant in the best molecular biology approaches for study of nitric oxide signaling and uncoupling. The applicant will benefit from multi-disciplinary mentorship within the Emergency Resuscitation Center, building upon expertise in heart oxidant injury and adaptation, and free radical biology. The sponsors Drs. Vanden Hoek and McNally are experienced physician-scientists. They are also collaborators interested in oxidantmediated heart contractile dysfunction and death in cardiomyocyte and mouse models. This Career Development Project builds upon the strengths of the applicant, the training program and institutional support and mentorship. We hypothesize that protective hypothermia following cardiac arrest increases Akt-related constitutive nitric oxide synthase (cNOS) NO-signaling, while attenuating cNOS uncoupling and generation of damaging reactive oxygen species (ROS). Specifically, in a mouse model of cardiac arrest we will: (Aim 1) Optimize the target temperature of protective hypothermia based on heart Akt and cNOS activity; (Aim 2) Test the relative importance of NOS1/NOS3 in post-cardiac arrest injury, NO and ROS generation, and hypothermia protection using selective-inhibitor and knockout strategies; and (Aim 3). Test whether hypothermia protection can be reproduced by NOS substrate/cofactor repletion strategies. The University of Chicago Department of Medicine and Section of Emergency Medicine have a track-record of successful mentoring using the K08 physician-scientist training program. The proposed training will provide Dr. Beiser with additional skills to become a leading independent physician-scientist in his field. From a layperson perspective, cardiac arrest (when the heart stops) is different than depicted on TV or the movies-most people initially resuscitated do not survive. This work will help understand how cooling the body a few degrees can actually increase a type of oxidant stress that helps the body heal itself.