Automated assays of gap junction functions and screening of drugs for rescuing conduction in heart tissues

心脏性猝死(SCD)占心脏病死亡人数的50%。SCD的主要原因是遗传 不良生活方式/肥胖引起的问题(从出生就存在)和缺血性心脏病，两者都会危及生命 心脏跳动不规律(心律不齐)。我们感兴趣的是致命性心律失常的遗传原因以及它们如何 是可以治疗的。通常，SCD的遗传原因会导致缝隙连接的变化，缝隙连接是一种特殊的通道， 允许心脏细胞之间的交流，促进心脏的协调收缩。新发现 这种疾病的治疗是具有挑战性的，因为测量缝隙连接的变化(和挽救)在技术上是困难的。 我们之前设计并发表了测量缝隙连接活性的新方法，并展示了缝隙是如何 结合部会因疾病而改变。我们的自动化系统将一种小的荧光染料注入单个心肌细胞， 并跟随染料的扩散来测量缝隙连接功能。在这个项目中，我们将进一步开发我们的系统，以 在96孔板结构中进行微量注射，使我们能够筛选和测试许多药物化合物的能力 来纠正这种疾病。此外，我们将设计一种新的基于细胞间钙移动的分析方法来验证和 将我们的结果关联起来，也是以96孔的形式。任何已确定的潜在疗法都将在两个人类心肌细胞中进行测试 心源性猝死的模型，显示这些药物是否很有可能对患者有效。我们将测试的大多数药物是 已经得到监管机构的批准，确保快速转化为患者护理。总而言之，我们的工作将设计出新的 以及测量心脏功能和为这种流行疾病的患者提供新疗法的新技术。

Sudden cardiac death (SCD) accounts for up to 50% of lives lost from heart disease. The major causes of SCD are genetic problems (present from birth) and ischemic heart disease resulting from poor lifestyle/obesity, both causing life-threatening irregular beating of the heart (arrhythmias). We are interested in the genetic causes of the lethal arrhythmias and how they can be treated. Often the genetic causes of SCD result in changes to gap junctions, which are specialized channels that allow for communication between heart cells and facilitate coordinated contraction of the heart. Discovery of new therapeutics for this condition is challenging since measuring changes to (and rescue of) gap junctions is technically difficult. We have previously devised and published novel methods to measure gap junction activity and have shown how gap junctions are modified with disease. Our automated system injects a small fluorescent dye into individual cardiomyocytes, and follows the spread of the dye to measure gap junction function. In this project, we will further develop our system to perform microinjection in a 96-well plate configuration, allowing us to screen and test many drug compounds for their ability to correct the disease. Additionally, we will devise a new assay based on calcium movement between cells to validate and correlate our results, also in a 96-well format. Any potential therapeutics identified will be tested in two human cardiomyocyte models of sudden cardiac death, showing if the drugs are highly likely to work in patients. Most of the drugs we will test are already approved by regulatory agencies, ensuring rapid translation to patient care. In summary, our work will devise new and novel techniques for measuring heart function and deliver new therapies to patients with this prevalent disease.