Method Profile
Endothelin-1 (ET-1)-induced remodeling is assessed by gene expression, patch clamp and video-motion analysis to test selective responses to Cav1.3 C-terminal delivery.
Compare intervention effects on repolarization, ion currents and motion-derived contraction in a human heart-failure-like cell model.
Cardiomyocyte monolayers derived from one male and one female human induced pluripotent stem cell (iPSC) line.
ET-1 exposure at 10 nM for 48 hours from day 20; the intervention group receives AAV6-Cav1.3 C-terminal delivery for 48 hours from day 22. Untreated, ET-1 and intervention groups are assessed on day 30.
Human cardiomyocytes and remodeling stimulus
Cardiomyocytes derived from two hiPSC lines; ET-1 induces molecular, electrical and contraction changes.
Gene-delivery component
AAV6 carries the human Cav1.3 C-terminal fragment spanning amino acids 1840–2137.
Functional measurements
RT-qPCR, single-cell patch clamp and phase-contrast particle tracking; video supplies contraction-amplitude measurements.
Evaluate molecular, single-cell electrical and monolayer-motion endpoints separately, retaining persistent sodium-current, upstroke and hypertrophy-related changes.
Parallel three-group comparison of ET-1 remodeling and the response to C-terminal delivery.
Two cell lines; action-potential recording n = 42/46/50 for untreated, ET-1 and intervention groups. Electrophysiology/contraction captions specify three independent differentiations; the expression figure separately specifies four biological replicates.
Untreated cells, ET-1 alone and ET-1 plus the C-terminal fragment.
ET-1 prolonged APD90/APD50 by 34%/17%. Fragment delivery restored repolarization, measured potassium/calcium currents and video contraction amplitude; sodium-current, upstroke and capacitance abnormalities persisted.
A preprint reports recovery of selected calcium and potassium currents, repolarization and video-derived contraction after delivery of a Cav1.3 fragment. Sodium-current and hypertrophy-related changes persisted.
