scaRNA20 Promotes Pseudouridylatory Modification of Small Nuclear snRNA U12 and Improves Cardiomyogenesis
Document Type
Article
Publication Title
Experimental Cell Research
Abstract
Non-coding RNAs, particularly small Cajal-body associated RNAs (scaRNAs), play a significant role in spliceosomal RNA modifications. While their involvement in ischemic myocardium regeneration is known, their role in cardiac development is unexplored. We investigated scaRNA20's role in iPSC differentiation into cardiomyocytes (iCMCs) via overexpression and knockdown assays. We measured scaRNA20-OE-iCMCs and scaRNA20-KD-iCMCs contractility using Particle Image Velocimetry (PIV), comparing them to control iCMCs. We explored scaRNA20's impact on alternative splicing via pseudouridylation (Ψ) of snRNA U12, analyzing its functional consequences in cardiac differentiation. scaRNA20-OE-iPSC differentiation increased beating colonies, upregulated cardiac-specific genes, activated TP53 and STAT3, and preserved contractility under hypoxia. Conversely, scaRNA20-KD-iCMCs exhibited poor differentiation and contractility. STAT3 inhibition in scaRNA20-OE-iPSCs hindered cardiac differentiation. RNA immunoprecipitation revealed increased Ψ at the 28th uridine of U12 RNA in scaRNA20-OE iCMCs. U12-KD iCMCs had reduced cardiac differentiation, which improved upon U12 RNA introduction. In summary, scaRNA20-OE in iPSCs enhances cardiomyogenesis, preserves iCMC function under hypoxia, and may have implications for ischemic myocardium regeneration.
DOI
10.1016/j.yexcr.2024.113961
Publication Date
3-1-2024
Keywords
Cardiomyocytes, Differentiation, Induced pluripotent stem cells, Pseudouridylation, Small Cajal-body associated RNA
ISSN
1090-2422
Recommended Citation
Perales S, Sigamani V, Rajasingh S, Gurusamy N, Bittel D, Czirok A, Radic M, Johnson R. scaRNA20 Promotes Pseudouridylatory Modification of Small Nuclear snRNA U12 and Improves Cardiomyogenesis. Experimental Cell Research. 2024; 436(1). doi: 10.1016/j.yexcr.2024.113961.
