Disruption of Autonomic Pathways in the Left Atrium by Inhibition of G-proteins

说明(申请人提供)：房颤(房颤)是最常见的心脏节律紊乱，是导致严重疾病的主要原因，如充血性心力衰竭和脑血管栓塞(中风)。重要的是，这种心律失常的发生率随着年龄的增长而增加，其结果是房颤正迅速成为老龄化人口中最新的“流行病”。因此，房颤的诊断和治疗已成为心血管医学的一个重要而具有挑战性的方面。然而，有效治疗房颤的进展缓慢，这在很大程度上是由于对这种心律失常的潜在机制缺乏了解。在这方面，最近的研究表明肺静脉和左后心房在这种心律失常的发生中起着重要的作用。因此，解放军已经进行了几次开创性的消融手术，尽管成功与否参差不齐。在心脏中，G蛋白偶联受体(GPCRs)及其同源信号伙伴异源三聚体G蛋白调节大多数机械和电功能。自主神经系统调节关键的心脏参数，如兴奋性、心率、收缩力、传导速度和不应性。与G1s相连的2-肾上腺素能受体的激活导致心脏传导速度和其他几种兴奋性反应的增加。与G1i偶联的M_2受体的激活，导致心房不应期显著缩短。结合起来，自主神经系统的这两个分支已经被证明为房颤创造了底物。因此，肾上腺素能和毒扁豆碱受体或它们的伴侣G1i和G1s可能是旨在调节心脏致心律失常影响的治疗策略的可行替代靶点。解放军可能是这些战略的一个特别有吸引力的目标，因为它具有非常强大和独特的自主神经特征，被认为有利于房颤。为了尝试修饰房颤的底物，我们建议使用针对GPCR/G蛋白界面的新型多肽来选择性地抑制解放军中的副交感或交感通路。使用可以在短期和长期基础上表达这些G蛋白抑制肽的微型基因(质粒)，拟议的研究将在急性和慢性房颤模型中进行。在急性实验中(目标1)，将迷走神经或肾上腺素介导的房颤抑制正常犬的房颤，将迷走神经或肾上腺素局部注射到解放军内。在目标2中，我们建议将这些迷你基因用于慢性房颤的犬模型；在长效启动子的控制下，将这些迷你基因局部注射到解放军中，以防止房颤自主神经底物的发展。拟议的研究是朝着确定最终可能应用于治疗危及生命的心律失常的新疗法迈出的重要一步。公共卫生相关性：房颤(AF)是最常见的心脏节律紊乱，是充血性心力衰竭和中风等严重疾病的主要原因。然而，目前可用的治疗房颤的方法并不是很有效。我们提出了一种新的治疗房颤的方法，通过使用一种新的肽(蛋白质)来抑制触发这种心律失常的神经功能。

DESCRIPTION (provided by applicant): Atrial fibrillation (AF) is the commonest rhythm disturbance of the heart, and is a major cause of serious morbidity such as congestive heart failure and cerebrovascular embolism (`stroke'). Importantly, the incidence of this arrhythmia increases with age, with the result that AF is fast becoming the latest `epidemic' in an aging population. The diagnosis and management of AF have therefore become an important and challenging aspect of cardiovascular medicine. However, progress in effectively treating AF has been slow, in large part due to a poor understanding of the underlying mechanisms of this arrhythmia. In this regard, recent studies indicate an important role for the pulmonary veins and the posterior left atrium (PLA) in the genesis of this arrhythmia. Several pioneering ablative procedures have therefore been performed in the PLA, albeit with mixed success. In the heart, G protein coupled receptors (GPCRs) and their cognate signaling partners, the heterotrimeric G-proteins, regulate most mechanical and electrical functions. The autonomic nervous system regulates critical cardiac parameters such as excitability, heart rate, force of contraction, conduction velocity and refractoriness. Activation of 2-adrenergic receptors, which are coupled to G1s, leads to an increase in conduction velocity and several other excitatory responses in the heart. Activation of muscarinic M2 receptors, which are coupled to G1i, leads to a marked shortening of refractoriness in the atria. In combination, these two limbs of the autonomic nervous system have been demonstrated to create substrate for AF. Thus, the adrenergic and muscarinic receptors or their partners G1i and G1s may be viable alternative targets for therapeutic strategies designed to modulate arrhythmogenic influences in the heart. The PLA may be an especially attractive target for these strategies, on account of a very robust and unique autonomic profile that is thought to be conducive to AF. In an attempt to modify substrate for AF, we propose to use novel peptides directed at the GPCR/G protein interface to selectively inhibit parasympathetic or sympathetic pathways in the PLA. Using minigenes (plasmids) that can express these G-protein inhibitory peptides on both a short and long term basis, the proposed studies will be performed in both an acute as well as a chronic model of AF. In the acute experiments (Aim 1), localized injection of minigene into the PLA will be performed in order to inhibit vagally or adrenergically-mediated AF in normal dogs. In Aim 2, we propose to use these minigenes in a canine model of chronic AF; minigenes under the control of a long-acting promoter will be injected locally into the PLA, to prevent the development of autonomic substrate for AF. The proposed studies are an important stride towards identifying novel therapeutics that may eventually be applied to the treatment of life threatening arrhythmias. PUBLIC HEALTH RELEVANCE: Atrial fibrillation (AF) is the commonest rhythm disturbance of the heart, and is a major cause of serious morbidity such as congestive heart failure and stroke. However, currently available treatment options for AF are not very effective. We propose a new method to treat AF, by using a novel peptide (protein) to inhibit the function of the nerves that trigger this arrhythmia.