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Description

Cardiovascular diseases remain a leading cause of mortality due to cardiomyocytes' inability to proliferate and regenerate after injury. Current therapies that broadly express regenerative factors in the heart are ineffective. However, targeted expression of these factors to the injury can induce cardiomyocyte proliferation. Our research introduces an innovative technique that can deliver regenerative factors directly to the injured myocardium using a biomarker uniquely expressed by injured cardiomyocytes, Natriuretic Peptide B (Nppb). Our research utilizes the CellREADR system, an ADAR-based RNA sensor that detects Nppb expression and induces translation of regenerative cardiac factors. The overall goal of this study was to identify the most efficient sensor. In the unamplified construct, all transfected cells express BFP (blue fluorescent protein). When Nppb mRNA is present, it binds to the sensor and recruits the ADAR enzyme, allowing RNA editing and GFP (green fluorescent protein) translation. The amplified construct incorporates a tetracycline-controlled transcriptional activation system to enhance GFP expression. HEK293T cells were transfected with these constructs and Nppb coding sequence, then sensor efficiency and specificity were assessed by counting GFP+ cells in experimental and control groups. The unamplified sensor successfully detected Nppb with minimal background activity. However, the amplified sensor showed unintended GFP expression in negative controls, indicating non-specific expression. Optimization efforts are underway to refine specificity by incorporating additional upstream regulatory elements. Our findings demonstrate the potential of RNA sensors in targeting injured cardiomyocytes, contributing to gene-based strategies for cardiac regeneration and repair.

Publication Date

4-1-2025

Keywords

cardiomyocyte regeneration, heart repair, Nppb biomarker, CellREADR system, ADAR-based RNA sensor, RNA editing, gene therapy, cardiac injury, B-type natriuretic peptide, HEK293T cells, GFP expression, sensor specificity, cardiac gene delivery, myocardial infarction, regenerative medicine, RNA-based therapeutics, cardiac-specific targeting, translational research, cardiac regeneration strategies, RNA sensor optimization.​

Targeting Injured Cardiomyocytes with RNA Sensors

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