High Throughput Screening of Pharmaceuticals Targeting Heart Cell Contractile Function

分离的心肌细胞代表了心肌最小的全功能模型系统。 心肌细胞对电刺激的反应而收缩，并有助于确定心脏的特征 在收缩、钙离子调节和动作电位方面起作用。目前， 可以使用当前技术检查和分析的心肌细胞数量在 数量级为每10分钟1个肌细胞。然而，为了适应更快的药物，需要更高的吞吐量 在基础研究方面的发现和日益增长的动力，朝着大规模快速数据收集和 分析。这笔赠款的重点是验证全自动身份识别的可行性。 非重叠存活心肌细胞的研究及收缩能力和钙离子调节的量化 在10分钟内，至少有100个心肌细胞。在制药环境中，这是一个重要的 增强表型量化，加快药物疗效/毒性评估，并 将允许公司消除更危险或更边缘的风险，从而大大降低成本 化合物。在目标1中，我们将开发一种轻量级的x-y可移动倒置显微镜，它将扫描 一片分离的心肌细胞。目标1的目标是识别分离的和可存活的心脏 根据肌细胞的形态和肌节模式，至少有100个可用肌细胞 在10分钟内确认。在目标2中，一种新图像分析方法将量化位置、大小 所有离体心收缩-松弛功能的定位和动态特征 视野内的肌细胞，同样以10分钟内100个肌细胞为目标。 IonOptix位于马萨诸塞州，提供了基于显微镜的设备用于评估 近25年来细胞内钙调节和收缩/松弛功能。

The isolated cardiac myocyte represents the smallest fully functional model system of heart muscle. Cardiac myocytes contract in response to electrical stimulation and are useful to characterize heart function in terms of contractility, calcium ion regulation, and action potential. Currently, the number of cardiac myocytes that can be examined and analyzed using current techniques is on the order of 1 myocyte per 10 min. Higher throughput is required, however, to accommodate faster drug discovery and a growing motivation in basic research toward large scale rapid data collection and analysis. This grant is focused on validating the feasibility of fully-automated identification of non-overlapping viable myocytes and quantification of contractility and calcium ion regulation in at least 100 myocytes in 10 min. In the pharmaceutical setting, this is a significant enhancement in phenotype quantification that expedites assessment of drug efficacy/toxicity and will greatly reduce costs by permitting companies to eliminate more dangerous or marginal compounds. In Aim 1, we will develop a lightweight x-y moveable inverted microscope that will scan a field of isolated cardiac myocytes. The goal of Aim 1 is to identify isolated and viable cardiac myocytes by their morphology and sarcomeric pattern such that at least 100 useable myocytes are identified within 10 min. In Aim 2, a novel image analysis method will quantify position, size, orientation and dynamic characteristics of contraction-relaxation function for all isolated cardiac myocytes within the field of view, again with the goal of 100 myocytes within 10 min. IonOptix is located in Massachusetts and has provided microscopy-based equipment for assessing intracellular calcium regulation and contraction/relaxation function for the last 25 years.