Role of the TRPV1 channel in myocardial salvage from ischemia-reperfusion injury

我是一名麻醉师，希望成为一名临床科学家。我感兴趣的是研究瞬时受体香草素通道（TRPV1）在缺血-再灌注损伤心肌修复中的作用。作为一名临床麻醉师，我觉得我有一套独特的技能，可以在基础科学实验室研究这个具体而相关的临床问题。我目前的研究方向是我之前关于心肌缺血再灌注损伤的基础科学研究的逻辑延续，并补充了我的临床训练。然而，距离我上一次在配体诱导的心脏保护方面的强化基础科学培训已经过去了5年，所以我的近期目标是获得先进分子生物学技术的额外研究培训，并专注于职业发展，以便向独立研究者过渡。额外的研究和职业培训将是无价的，并使我能够回答相关的问题，否则我无法研究。在该奖项的支持下，这项工作将为建立麻醉学教师、临床医生和基础科学家的职业生涯奠定基础。培训环境：斯坦福大学拥有一流的职业发展环境，以及从事和学习新颖、前沿研究的环境。斯坦福大学致力于让我成为一名临床科学家，这一点从医学院、心血管研究所、麻醉学系、化学和系统生物学系、我的导师（Daria Mochly-Rosen博士）、共同导师（Rona Giffard博士和David Yeomans博士）以及他们的实验室成员对我的建议培训中就可以看出来。我组建了一支在转化研究、蛋白质化学与生物化学、心肌与神经元缺血再灌注损伤与疼痛研究方面具有多学科研究专长和技能的咨询团队。研究生涯发展的关键要素包括提高我在蛋白质生物化学，蛋白质-蛋白质相互作用和疼痛信号分子机制方面的知识。我还将在基于细胞的技术和小分子药物开发和设计方面获得额外的培训。研究：辣椒产生辛辣的味道是由复合辣椒素产生的，辣椒素反过来也会释放内啡肽（这可能是我们喜欢辛辣食物的原因）。辣椒素敏感通道的激活，被称为瞬时受体电位香草素1 (TRPV1)，可能不仅仅是疼痛和随后的快乐的信号。在心脏病发作时，TRPV1可能激活一种信号通路，减少因缺氧对心脏（缺血）造成的损伤。这可能表明，胸痛（心绞痛）实际上是防止心脏病发作的自然途径。由TRPV1激活产生的痛觉是否与防止心脏病发作的途径有关？糖尿病会增加心脏病发作的心血管风险，从而降低TRPV1的表达。糖尿病患者也有无声或无痛的心脏病发作。没有疼痛（TRPV1激活），是否没有获得器官保护？研究疼痛和保护之间的联系也很重要，因为通过阻断TRPV1通道来减轻疼痛的药物正在临床试验中。在本提案中，我将展示TRPV1通道如何减少心脏病发作造成的损伤。我将通过研究TRPV1糖基化（Aim 1）来确定分子基础，其次，研究蛋白激酶C ε silon （PKC5）磷酸化（Aim2）是否减轻心脏细胞损伤。然后，我将检查通常在手术室使用的药物，阿片类药物和挥发性麻醉剂，是否需要TRPV1来减少缺血再灌注损伤的损害（目的3）。我还将确定在糖尿病患者中，阿片类药物和挥发性麻醉剂对心脏病发作的保护作用的丧失是否可以通过改善TRPV1的敏感性来逆转（Aim 4）。总之，这些研究将确定胸痛和心脏病发作损伤之间的联系，以及阿片类药物和挥发性麻醉剂如何通过激活TRPV1启动减少心脏病发作损伤的途径。

DESCRIPTION (provided by applicant): I am an anesthesiologist that desires a career as a clinician scientist. I am interested in examining the role of the transient receptor vanilliod channel (TRPV1) in myocardial salvage from ischemia- reperfusion injury. As a clinical anesthesiologist, I feel I have a skill set unique to studying this specific and pertinent clinical question in the basic science laboratory. My current research direction is a logical continuation from my previous basic science research regarding myocardial ischemia-reperfusion injury and compliments my clinical training. However, 5 years have passed since my last intensive basic science training in ligand-induced cardioprotection, so my immediate goal is to obtain additional research training in advanced molecular biology techniques and focused career development in order to transition to an independent investigator. Additional research and career training will be invaluable and allow me to answer pertinent questions that I could not examine otherwise. This work, with support by this award, will provide a foundation to establish a career as a teacher, clinician and a basic scientist in Anesthesiology. Training Environment: Stanford has an outstanding world-class environment for career development and to perform and learn novel, cutting edge research. Stanford is committed to my development as a clinician scientist, as evident by my proposed training from members in the School of Medicine, the Cardiovascular Research Institute, the Department of Anesthesiology, Department of Chemical and Systems Biology, my mentors (Dr. Daria Mochly-Rosen), co-mentors (Dr. Rona Giffard and Dr. David Yeomans) and their lab members. I assembled an advisory team with multidisciplinary research expertise and skills in translational research, protein chemistry and biochemistry, myocardial and neuronal ischemia-reperfusion injury and pain research. Key elements for research career development include enhancing my knowledge of protein biochemistry, protein-protein interactions and the molecular mechanisms of pain signaling. I will also obtain additional training in cellular-based techniques and small molecule drug development and design. Research: Chili peppers produce their hot spicy taste by the compound capsaicin, which in turn also releases endorphins (a probable reason why we are fond of spicy food). Activation of the capsaicin-sensitive channel, known as the transient receptor potential vanilloid 1 (TRPV1), may signal more than just pain and subsequent pleasure. TRPV1 may activate a signaling pathway which reduces injury from a lack of oxygen to the heart (ischemia) during a heart attack. This may suggest that the chest pain (angina) actually is a natural pathway to protect from heart attack injury. Is the pain sensation generated by TRPV1 activation linked to a pathway which protects from heart attacks? Diabetes, which increases cardiovascular risk for a heart attack, reduces TRPV1 expression. Diabetics also have heart attacks that are silent or painless. With no pain (TRPV1 activation), is there no gain of organ protection? The connection between pain and protection is also important to study because drugs to reduce pain by blocking the TRPV1 channel are in clinical trials. In this proposal, I will show how the TRPV1 channel reduces injury from a heart attack. I will determine the molecular basis by studying whether TRPV1 glycosylation (Aim 1) and secondly, protein kinase C epsilon (PKC5) phosphorylation (Aim2), reduce heart cell injury. I will then examine whether agents commonly given in the operating room, opioids and volatile anesthetics, require TRPV1 to reduce damage from ischemia-reperfusion injury (Aim 3). I also will determine whether in diabetes the loss of protection from a heart attack by opioids and volatile anesthetics can be reversed by improving TRPV1 sensitivity (Aim 4). Together, these studies will identify how chest pain and heart attack injury are connected and how opioids and volatile anesthetics initiate a pathway by TRPV1 activation which reduces heart attack injury. PUBLIC HEALTH RELEVANCE: Project Narrative Over one million people annually in the United States suffer a heart attack. A spicy diet in moderation or modification of TRPV1 pain receptors may prevent or reduce the extent of heart attack injury for people at risk.