Abstract
Cardiac hypertrophy, a precursor to heart failure (HF), involves intricate signaling networks characterized by epidermal growth factor receptor (EGFR) activation and calcium (Ca(2+)) dysregulation. Therapeutic inhibition of EGFR has emerged as a promising approach to attenuate maladaptive hypertrophic remodeling, particularly by restoring Ca(2+) homeostasis, a critical factor in maintaining myocardial function. However, drug discovery targeting EGFR/Ca(2+) pathways remains constrained by the limited proliferative capacity of human cardiomyocytes (CMs) and the lack of real-time probes capable of concurrently monitoring EGFR and Ca(2+) signaling in living cells. To address these limitations, we developed a tetrahedral DNA nanostructure-based probe (TDN-EA) integrated with human embryonic stem cell-derived cardiomyocytes (hESC-CMs) for real-time, concurrent detection of EGFR and Ca(2+) dynamics via fluorescence resonance energy transfer (FRET)-ON mechanism. The TDN-EA probe demonstrated high specificity, stability, and biocompatibility in hESC-CMs. Leveraging TDN-EA, we established a high-throughput screening platform that identified paromomycin (PM) as a novel therapeutic candidate from a library of 420 natural compounds. PM attenuated cardiac hypertrophy effectively in vitro and in vivo by inhibiting EGFR/Ca(2+) signaling pathway. This study underscores the potential of TDN-EA as a transformative tool for high-throughput drug discovery, enabling the identification of therapeutics that simultaneously target EGFR and Ca(2+) signaling pathways in cardiac hypertrophy.