Method Profile
A patient-specific iPSC-cardiomyocyte model comparing CAV3 localization and calcium transient behavior with a relative's control cells.
Examine whether a defined human-cell background shows different baseline or fenoterol-stimulated calcium activity.
Cardiomyocytes derived from one CAV3 p.Ala46Thr carrier and an unaffected sister without the variant.
Protein imaging and ultrastructure assessment paired with Fluo-4 calcium recordings before and after fenoterol.
Cell identity
Patient-derived and control iPSC lines differentiated to cardiomyocytes; control was a relative, not a gene-corrected isogenic line.
Stimulus
Fenoterol, a beta-adrenergic agonist, compared with each cell group's baseline activity.
Measurements
CAV3 protein localization, cell structure, calcium transient frequency and interval variation.
Keep cellular and donor denominators separate; compare both frequency and regularity rather than treating a faster calcium signal as functional normalization.
Variant-carrier versus unaffected-sister cardiomyocytes, each measured before and after fenoterol.
One donor per genetic background; CAV3 localization in 23 patient-derived and 22 control cells; calcium recordings from 13 cardiomyocytes per group across at least three experiments.
Unaffected-sister cells and within-group unstimulated activity.
Abnormal CAV3 localization in 20/23 patient cells versus 0/22 controls. Fenoterol increased transient frequency from 2.8 to 4.4/min in patient cells, but interval variation rose from 22.73% to 31.42%; control frequency rose from 4.0 to 7.0/min with low interval variation.
Cardiomyocytes derived from one CAV3 p.Ala46Thr carrier showed altered protein localization and irregular calcium transients. Fenoterol increased transient frequency, but the pattern remained less regular than in a relative's control cells.
