Abstract
The role of long non-coding RNA (lncRNA) in endogenous cardiac regeneration remains largely elusive. The mammalian cardiomyocyte is capable of regeneration for a brief period after birth. This fact allows the exploration of the roles of critical lncRNAs in the regulation of cardiac regeneration. Through a cardiac regeneration model by apical resection (AR) of the left ventricle in neonatal mice, we identified an lncRNA named natriuretic peptide A antisense RNA 1 (NPPA-AS1), which negatively regulated cardiomyocyte proliferation. In neonates, NPPA-AS1 deletion did not affect heart development, but was sufficient to prolong the postnatal window of regeneration after AR. In adult mice, NPPA-AS1 deletion improved cardiac function and reduced infarct size after myocardial infarction (MI), associated with a significant improvement in cardiomyocyte proliferation. Further analysis showed that NPPA-AS1 interacted with DNA repair-related molecule splicing factor, proline- and glutamine-rich (SFPQ). A heteromer of SFPQ and non-POU domain-containing octamer-binding protein (NONO) was required for double-strand DNA break repair, but NPPA-AS1 was competitively bound with SFPQ due to the overlapped binding sites of SFPQ and NONO. NPPA-AS1 deletion promoted the binding of SFPQ–NONO heteromer, decreased DNA damage, and activated cardiomyocyte cell cycle re-entry. Together, loss of NPPA-AS1 promoted cardiomyocyte proliferation by stabilizing SFPQ–NONO heteromer-induced DNA repair and exerted a therapeutic effect against MI in adult mice. Consequently, NPPA-AS1 may be a novel target for stimulating cardiac regeneration to treat MI.
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UCSC database (http://genome-asia.ucsc.edu/index.html) was used to determine the chromosomal location of targeted lncRNA. Through Coding Potential Assessment Tool (CPAT) database (http://lilab.research.bcm.edu/cpat/index.php), the coding potential of NPPA-AS1 was predicted. The cDNA sequences of human- and mouse-NPPA-AS1 were compared by Global Align tools: https://blast.ncbi.nlm.nih.gov/Blast.cgi?PAGE_TYPE=BlastSearch&PROG_DEF=blastn&BLAST_PROG_DEF=blastn&BLAST_SPEC=GlobalAln&LINK_LOC=BlastHomeLink. RNA structure analysis was predicted by RNA fold (http://rna.tbi.univie.ac.at/cgi-bin/RNAWebSuite/RNAfold.cgi) and Forna websites (http://rna.tbi.univie.ac.at/forna/forna.html). PanglaoDB (https://panglaodb.se) was used as a database to explore the cellular distribution of lncRNA when the protocol of single-cell RNA sequencing was SMART-seq2. We performed an enrichment analysis of signal pathways through Metascape (http://metascape.org/gp/index.html#/main/step1). The structure of SFPQ/NONO was referenced from the database of Protein Data Bank (https://www.rcsb.org/structure/4WIJ). The nucleotide-binding proteome of lncRNA was estimated via algorithms provided by catRAPID omics (http://service.tartaglialab.com/update_submission/274908/a801f9d744); RNA-binding regions of SFPQ proteins and NPPA-AS1 were predicted by catRAPID fragments (http://service.tartaglialab.com/update_submission/274910/fe4a85fb9e).
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This work was supported by grants from the National Science Foundation of China (81930008, U20A20344), Program of Innovative Research Team by National Natural Science Foundation (81721001, 81922005), National Key Research & Development Program of China (2018YFA0107403), and Chongqing Natural Science Foundation (cstc2020jcyj-jqX0016).
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Fu, W., Ren, H., Shou, J. et al. Loss of NPPA-AS1 promotes heart regeneration by stabilizing SFPQ–NONO heteromer-induced DNA repair. Basic Res Cardiol 117, 10 (2022). https://doi.org/10.1007/s00395-022-00921-y
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DOI: https://doi.org/10.1007/s00395-022-00921-y