High Throughput Screening of Pharmaceuticals Targeting Heart Cell Contractile Function

针对心脏细胞收缩功能的药物的高通量筛选

• 批准号: 9347047
• 负责人: BRADLEY M PALMER
• 金额: $ 20.51万
• 依托单位: IONOPTIX, LLC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9347047
• 关键词: Action Potentials; Address; Affect; Attention; Automation; Basic Science; Bathing; Biological Models; Calcium; Calcium ion; Cardiac; Cardiac Myocytes; Cell physiology; Cell-Matrix Junction; Cells; Characteristics; Computer software; Contracts; Dangerousness; Data Analyses; Data Collection; Detection; Devices; Electric Stimulation; Equipment; Exposure to; Feasibility Studies; Fluorescent Dyes; Fourier Transform; Goals; Grant; Heart; Heart Diseases; Heart Research; Human Resources; Image Analysis; Individual; Ions; Laboratories; Liquid substance; Masks; Massachusetts; Measurement; Mechanics; Membrane Potentials; Methods; Microscope; Microscopy; Morphology; Motivation; Movement; Muscle; Muscle Cells; Myocardium; Pattern; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phenotype; Positioning Attribute; Preparation; Process; Regulation; Relaxation; Reporting; Reproducibility of Results; Resources; Risk; Sarcomeres; Scanning; System; Systems Analysis; Techniques; Toxic effect; Treatment Protocols; base; cost; disease phenotype; drug development; drug discovery; drug efficacy; fluid flow; heart cell; heart function; high throughput screening; innovation; ion dynamics; light weight; novel; programs; reagent testing; response; screening; statistics; two-dimensional; unconscious bias

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.

分离的心肌细胞代表心肌的最小的全功能模型系统。 心肌细胞响应于电刺激而收缩，并且可用于表征心脏 在收缩性、钙离子调节和动作电位方面起作用。目前 使用当前技术可以检查和分析的心肌细胞的数量是在 然而，需要更高的通量来适应更快的药物， 基础研究中的发现和日益增长的动力转向大规模快速数据收集， 分析.该补助金的重点是验证全自动化的可行性， 识别 非重叠存活肌细胞的数量以及收缩性和钙离子调节的定量 在药物环境中，这是一个显著的 表型定量的增强，加速了药物疗效/毒性的评估， 将大大降低成本，允许公司消除更危险或边际 化合物.在目标1中，我们将开发一种轻型x-y可移动倒置显微镜， 分离的心肌细胞的区域。目标1的目标是识别分离的和存活的心脏 通过它们的形态和肌节模式，使至少100个可用的肌细胞 在目标2中，一种新的图像分析方法将量化位置，大小， 离体心脏收缩-舒张功能的方向性和动态特性 在视野内的肌细胞，再次以在10分钟内100个肌细胞为目标。 IonOptix位于马萨诸塞州，提供基于显微镜的设备， 细胞内钙调节和收缩/舒张功能。