Mitochondrial K+ Channels and Cardioprotection

描述（由申请人提供）：我们研究计划的总体目标是阐明缺血再灌注（IR）损伤的内源性和其他保护机制，并利用这些知识开发IR疾病（如心脏病发作和中风）的新疗法。许多心脏保护策略似乎集中在线粒体钾通道作为保护信号的必要和充分的效应器。然而，这些渠道的身份和监管仍然存在争议。迄今为止，我们发表的研究和本文所包含的令人兴奋的初步数据将我们的重点放在了一种新型线粒体K+通道上，这种通道是保护所必需的，并且以前没有涉及保护信号。值得注意的是，缺乏该通道似乎会产生代谢表型。我们还确定了一类新的内源性通道调节剂。在这项提案中，目标1将描述通道及其在心脏保护中的作用，目标2将研究通道之间的联系。 通道和心脏代谢，目的3将研究内源性信号对其的调节。我们将使用各种最先进的技术，包括线粒体（分离的线粒体内膜）的膜片钳和Seahorse XF方法来评估心肌细胞的生物能量学。该双PI方案借鉴了两位研究者的专业知识（Brookes -线粒体生物学、代谢筛选、心脏病理生理学; Nehrke -离子通道、小鼠遗传学、线粒体生理学）。我们富有成效的跟踪记录（该项目在3年内资助了8篇原创研究文章和3篇综述）表明，这3个目标的完成很有可能会产生关于该通道的关键信息，这是一种新的潜在的心脏保护药物靶点。

DESCRIPTION (provided by applicant): The overall goal of our research program is to elucidate endogenous and other mechanisms of protection against ischemia-reperfusion (IR) injury, and to exploit this knowledge to develop new therapies for IR disease conditions such as heart attack and stroke. Many cardioprotective strategies appear to converge on mitochondrial potassium channels as necessary and sufficient effectors of protective signaling. However, the identity and regulation of these channels remains controversial. Our published research to date, and exciting preliminary data contained herein, have directed our focus to a novel mitochondrial K+ channel that is required for protection and has not previously been implicated in protective signaling. Notably, absence of this channel appears to yield a metabolic phenotype. We have also identified a novel class of endogenous channel modulators. In this proposal, Aim 1 will characterize the channel and its role in cardioprotection, Aim 2 will investigate links between the channel and cardiac metabolism, and Aim 3 will study its regulation by endogenous signals. We will use a variety of state-of-the -art techniques, including patch-clamp of mitoplasts (isolated mitochondrial inner membranes), and Seahorse XF methodology to assess cardiomyocyte bioenergetics. This dual-PI proposal draws on the expertise of both investigators (Brookes - mitochondrial biology, metabolic screening, cardiac patho-physiology; Nehrke - ion channels, mouse genetics, mitochondrial physiology). Our productive track-record (8 original research articles and 3 reviews funded by this project in 3 years) imparts a high probability that the completion of these 3 aims will yield critical information about this channel, which is a novel potential drug target for cardioprotection.