High Throughput Screening of Pharmaceuticals Targeting Heart Cell Contractile Function

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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.