High-throughput evaluation of cardiac-specific promoters for adeno-associated virus mediated cardiac gene therapy

作者：Dhanya Ravindran, Renuka Rao, Juan Mundisugih, Tracy Titus, Shinya Tsurusaki, Cindy Kok, Fairooj N. Rashid, Sindhu Igoor, Yasuhito Kotake, Saurabh Kumar, James J.H. Chong, Ian E. Alexander, Leszek Lisowski, Eddy Kizana · 发表于：Gene Therapy · 年份：2025 · DOI：10.1038/s41434-025-00553-6 · 被引用次数：7 · 研究领域：Viral Infections and Immunology Research、Virus-based gene therapy research、RNA Interference and Gene Delivery

The selection of an appropriate promoter is important to the design and optimisation of adeno-associated viral (AAV) vector-based cardiac gene therapies. The expression cassette design can impact efficacy and safety of the vector. This study is the first to use a novel AAV barcode-seq method for the simultaneous evaluation of a panel of cardiac-specific promoters in a high-throughput manner. Functional analyses of our cardiac promoter kit packaged in three different capsids were performed using neonatal rat ventricular myocytes (NRVM), human iPSC-derived cardiomyocytes (hiPSC-CMs), HuH7 hepatocellular carcinoma cells, as well as mouse, rat, sheep and pig models. The cardiac troponin T (cTnT) promoter showed the most promise overall as a cardiac-specific promoter across all cardiac models tested. The results validate the barcode-seq technique as a powerful and versatile approach that enables high-throughput, quantitative analysis of various expression cassettes in commonly used models of cardiac gene therapy.