Data from: Robotic manipulation of cardiomyocytes to identify gap junction modifiers for arrhythmogenic cardiomyopathy

Arrhythmogenic cardiomyopathy (ACM) is a leading cause of sudden cardiac
death among young adults. Aberrant gap junction remodeling has been linked
to disease-causative mutations in plakophilin-2 (PKP2). Although gap
junctions are a key therapeutic target, measurement of gap junction
function in preclinical disease models is technically challenging. To
quantify gap junction function with high precision and high consistency,
we developed a robotic cell manipulation system with visual feedback from
digital holographic microscopy (DHM) for three dimensional and label-free
imaging of human induced pluripotent stem cell-derived cardiomyocytes
(iPSC-CMs). The robotic system can accurately determine the dynamic height
changes in the cells’ contraction and resting phases, microinject
drug-treated healthy and diseased iPSC-CMs in their resting phase with a
constant injection depth across all cells, and deposits a
membrane-impermeable dye that solely diffuses between cells through gap
junctions for measuring the gap junction diffusion function. The robotic
system was applied towards a targeted drug screen to identify
 gap junction modulators and potential therapeutics for ACM. Five
compounds were found to dose-dependently enhance gap junction permeability
in cardiomyocytes with PKP2 knockdown. Additionally, PCO 400 (Pinacidil)
reduced beating irregularity in a mouse model of ACM expressing mutant
PKP2 (R735X). These results highlight the utility of the robotic cell
manipulation system to efficiently assess gap junction function in a
relevant preclinical disease model, thus providing a technique to advance
drug discovery for ACM and other gap junction-mediated diseases.