Electrical Silencing of the Pulmonary Veins

肺静脉电沉默

基本信息

批准号：
7787967
负责人：
David J Milan
金额：
$ 26.33万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-02-03 至 2012-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7787967
关键词：
Ablation Academic Medical Centers Address Animal Model Animals Anterior Anticoagulation Arrhythmia Atrial Fibrillation Biological Cardiac Myocytes Cardiac Tamponade Cell Survival Cells Complication Coupled Data Dose Epitopes Esophageal Fistula Evaluation Experimental Animal Model Family Family member Gene Delivery Gene Transfer Genes Goals Heart Atrium Heterogeneity Holter Electrocardiography Hour In Vitro Maps Membrane Potentials Methods Molecular Muscle Cells Nature Nerve Paralysis Neurons Pacemakers Play Potassium Potassium Channel Prevalence Procedures Proteins Pulmonary veins Radiofrequency Interstitial Ablation Rest Right atrial structure Risk Role Sheep Stenosis Stroke Structure Structure of phrenic nerve Technology Time Tissues Toxic effect Transfection Transient Ischemic Attack Work atrioventricular node cytotoxicity gene discovery gene therapy implantation in vivo novel strategies overexpression programs public health relevance radiofrequency response success

项目摘要

DESCRIPTION (provided by applicant): The recent discovery of genes responsible for the potassium leak currents, the KCNK family of potassium channels, has created the opportunity to directly target arrhythmogenic triggers of atrial fibrillation, rather than merely trying to contain them through pulmonary vein isolation. Expression of the KCNK0 gene in neurons can render these cells electrically silent by effectively shorting out any depolarizing currents. We have extended this work to demonstrate that expression of KCNK0 in cardiomyocytes renders them inexcitable. We thus propose to evaluate the KCNK0 gene for its ability to silence cardiomyocytes as a treatment for atrial fibrillation in the following specific aims: Specific Aim1: In vitro evaluation of KCNK0 for the electrical silencing of cardiomyocytes. This will involve assessment of biological effects in cardiomyocytes - establishing transfection efficiency targets, assessing cell viability, studying the electrophysiologic effects of heterogeneity from mosaic KCNK0 expression, and assessing possible cytotoxicity. Specific Aim 2: In vivo adenoviral delivery of KCNK0 to the anterior right atrium to create a line of block. This will involve linear delivery of the Ad.KCNK0 followed 5 days later by multielectrode recordings of electrical activity. Comparison of two gene delivery methods and dose response will be evaluated with assessment of gene transfer efficiency, toxicity, and protein levels. Specific Aim 3: In vivo adenoviral delivery of KCNK0 to silence pulmonary vein myocytes in a large animal model. This will involve baseline electroanatomic mapping, followed by KCNK0 gene delivery to the pulmonary veins, and then remapping of the pulmonary veins to document electrical silencing. Animals will also undergo atrial programmed stimulation and Holter monitoring to assess for proarrhythmia. Comparison will be made to Ad.GFP treated animals as well as animals treated with traditional radiofrequency pulmonary vein isolation. PUBLIC HEALTH RELEVANCE: Our primary objective is to demonstrate the feasibility of electrically silencing the pulmonary veins using gene transfer as a potential treatment for atrial fibrillation in an experimental animal model. In this project, sheep be will be used to determine whether expression of a modified potassium leak current, KCNKO, by adenoviral gene therapy approach leads to loss of electrical excitability (electrical silencing) of the cardiomyocytes investing the pulmonary vein ostia.

描述（由申请人提供）：最近发现负责钾泄漏电流的基因，KCNK的钾通道家族创造了机会直接靶向房颤的心律失常触发因素，而不仅仅是试图通过肺静脉隔离来遏制它们。 KCNK0基因在神经元中的表达可以通过有效地缩短任何去极化电流来使这些细胞产生电沉降。我们已经扩展了这项工作，以证明KCNK0在心肌细胞中的表达使它们变得不可避免。因此，我们建议评估KCNK0基因在以下特定目的中静音心肌细胞的能力：特定目标：特定AIM1：对KCNK0的体外评估，用于对心肌细胞的电沉默。这将涉及评估心肌细胞中的生物学作用 - 建立转染效率靶标，评估细胞活力，研究从马赛克KCNK0表达中异质性的电生理效应，并评估可能的细胞毒性。特定目标2：在体内腺病毒在前右心房的腺病毒递送以创建块。这将涉及AD.KCNK0的线性传递，然后在5天后通过电活动的多电极记录。通过评估基因转移效率，毒性和蛋白质水平的评估，将评估两种基因递送方法和剂量反应的比较。特定目的3：在大型动物模型中，kcnk0的体内腺病毒递送，以使肺静脉肌细胞保持沉默。这将涉及基线电工映射，然后涉及KCNK0基因递送至肺静脉，然后重新映射肺静脉以记录电沉降。动物还将接受心房编程的刺激和抛光刺激，以评估心律失常。将与AD.GFP处理的动物以及用传统射频肺静脉分离进行比较。公共卫生相关性：我们的主要目标是证明使用基因转移将肺静脉电气沉默的可行性，作为在实验动物模型中对房颤的潜在治疗方法。在该项目中，将使用绵羊来确定通过腺病毒基因治疗方法的改性钾泄漏电流KCNKO的表达是否会导致投资肺静脉ostia的心肌细胞的电兴奋性（电沉降）。