MOLECULAR PHARMACOLOGY OF AN INHERITED HEART DISEASE

描述（改编自申请人的摘要）： 本申请中提出的研究是开发新的治疗药物 方法，基于遗传分子遗传的特定特性， 缺陷，以管理发生在 一种遗传性心脏病的两种形式（LQT-3和LQT-1），长QT间期 综合征 该项目旨在整合临床、分子和 细胞研究，以测试整体假设，突变， 编码心脏钠通道α亚基（SCN 5A）的基因，以及 慢钾通道电流（IKs）KvLQT-1/或minK原因可识别 表达的钠和钾通道活性的变化， 复极和相关节律的疾病相关变化 干扰，这反过来又使突变通道成为不同的目标， 治疗药物 因此，本研究的长期目标是 开发更有效和更具体的治疗方法来管理和 预防与这种疾病相关的危及生命的心律失常， 将开发针对特定基因缺陷的疗法。 在 将使用膜片钳程序进行体外实验， 测量人胚肾细胞中表达的全细胞电流 （HEK 293）和中国仓鼠卵巢（CHO）细胞，其已被瞬时转染。 用编码野生型（hH 1）和LQT-3突变体（deltaKPQ）的cDNA转染 人钠通道α亚单位的形式以及具有 用编码野生型和突变型KvLQT 1的cDNA共转染 还有Mink 着重于肾上腺素能调节的可能作用的实验， 细胞pH和钙内流将测试电压依赖性动力学， 可能区分KvLQT-1与SCN 5A衍生的神经体液因子 表型。 此外，还将对每个基因进行实验 针对设计的特定药理干预措施进行缺陷测试 以补偿的方式调节表达的通道活动 个体基因缺陷 主要研究者将咨询Dr. 罗切斯特大学的亚瑟·J·莫斯将执导 平行临床研究，以优化药理学方法， 管理和纠正已识别的基因缺陷。 实验数据来自 重组通道活性将与结果共享和整合 临床非侵入性心电学研究， LQT-1和LQT-3基因突变携带者与非携带者的体内研究 优化实验设计和治疗方法。

DESCRIPTION (adapted from the applicant's abstract): The overall goal of the research proposed in this application is to develop novel therapeutic approaches, based on specific properties of an inherited molecular genetic defect, to the management of electrophysiological aberrations that occur in two forms (LQT-3 and LQT-1) of an inherited cardiac disorder, the long QT syndrome. This project is designed to integrate clinical, molecular, and cellular studies in order to test the overall hypothesis that mutations in genes that encode the heart sodium channel alpha-subunit (SCN5A), and the slow potassium channel current (IKs) KvLQT-1/or minK cause identifiable changes in expressed sodium and potassium channel activity that underlie diseased-associated changes in repolarization and associated rhythm disturbances and that, in turn, make mutant channels distinct targets of therapeutic drugs. Thus, it is the long-term goal of this research to develop a more effective and specific therapeutic approach to manage and prevent life-threatening arrhythmias associated with this disease and that therapies will be developed that are targeted for specific gene defects. In vitro experiments will be carried out using patch-clamp procedures to measure whole-cell currents expressed in human embryonic kidney cells (HEK293) and Chinese hamster ovary (CHO) cells that have been transiently transfected with cDNAs encoding wild-type (hH1) and LQT-3 mutant (deltaKPQ) forms of the human sodium channel alpha-subunit as well as cells that have been co-transfected with cDNA encoding wild-type and mutant forms of KvLQT1 and minK. Experiments focusing on possible roles of adrenergic modulation, cellular pH and calcium influx will test for voltage-dependent kinetic and neurohumoral factors that may distinguish KvLQT-1 from SCN5A-derived phenotypes. In addition, experiments will be carried out on each gene defect testing for specific pharmacological interventions that are designed to modulate expressed channel activity in a manner to compensate for individual gene defects. The principal investigator will consult with Dr. Arthur J. Moss at the University of Rochester, who will be directing parallel clinical studies in order to optimize pharmacological approaches to manage and correct identified gene defects. Experimental data obtained from recombinant channel activity will be shared and integrated with the results of clinical non-invasive electrocardiologic studies that will be carried out in vivo on carriers vs. non-carriers of the LQT-1 and LQT-3 gene mutations to optimize experimental design and therapeutic approaches.