High-throughput screening of EGFR/Ca²⁺ signaling modulators in cardiac hypertrophy using a tetrahedral DNA...

Area of Science:

• Biochemistry
• Cardiovascular Biology
• Nanotechnology

Background:

• Cardiac hypertrophy precedes heart failure, driven by complex signaling networks involving epidermal growth factor receptor (EGFR) and calcium (Ca²⁺) dysregulation.
• Inhibiting EGFR shows promise for treating cardiac hypertrophy by restoring Ca²⁺ homeostasis, crucial for heart function.
• Current drug discovery is limited by low cardiomyocyte proliferation and the absence of real-time probes for simultaneous EGFR and Ca²⁺ signaling monitoring.

Purpose of the Study:

• To develop a novel probe for real-time, concurrent monitoring of EGFR and Ca²⁺ signaling in cardiomyocytes.
• To establish a high-throughput screening platform for identifying therapeutics targeting the EGFR/Ca²⁺ pathway.
• To identify novel therapeutic candidates for cardiac hypertrophy.

Main Methods:

• Development of a tetrahedral DNA nanostructure-based probe (TDN-EA) for fluorescence resonance energy transfer (FRET)-ON detection.
• Integration of TDN-EA with human embryonic stem cell-derived cardiomyocytes (hESC-CMs) for live-cell imaging.
• High-throughput screening of 420 natural compounds using the TDN-EA platform.

Main Results:

• The TDN-EA probe exhibited high specificity, stability, and biocompatibility in hESC-CMs.
• Paromomycin (PM) was identified as a novel therapeutic candidate that effectively attenuated cardiac hypertrophy.
• PM demonstrated therapeutic efficacy in vitro and in vivo by inhibiting the EGFR/Ca²⁺ signaling pathway.

Conclusions:

• The TDN-EA probe is a transformative tool for real-time, concurrent monitoring of EGFR and Ca²⁺ dynamics.
• The developed platform enables efficient high-throughput drug discovery for cardiac hypertrophy.
• Paromomycin represents a promising therapeutic agent for cardiac hypertrophy targeting the EGFR/Ca²⁺ pathway.