Identification and Initial Functional Characterization of a Human Vascular Cell-Enriched Long Noncoding RNA

Arteriosclerosis Thrombosis and Vascular Biology (Impact Factor: 6). 02/2014; 34(6). DOI: 10.1161/ATVBAHA.114.303240
Source: PubMed

ABSTRACT Long noncoding RNAs (lncRNAs) represent a rapidly growing class of RNA genes with functions related primarily to transcriptional and post-transcriptional control of gene expression. There is a paucity of information about lncRNA expression and function in human vascular cells. Thus, we set out to identify novel lncRNA genes in human vascular smooth muscle cells and to gain insight into their role in the control of smooth muscle cell phenotypes.
RNA sequencing of human coronary artery smooth muscle cells revealed 31 unannotated lncRNAs, including a vascular cell-enriched lncRNA (smooth muscle and endothelial cell-enriched migration/differentiation-associated long noncoding RNA [SENCR]). Strand-specific reverse transcription polymerase chain reaction (PCR) and rapid amplification of cDNA ends indicate that SENCR is transcribed antisense from the 5' end of the FLI1 gene and exists as 2 splice variants. RNA fluorescence in situ hybridization and biochemical fractionation studies demonstrate SENCR is a cytoplasmic lncRNA. Consistent with this observation, knockdown studies reveal little to no cis-acting effect of SENCR on FLI1 or neighboring gene expression. RNA-sequencing experiments in smooth muscle cells after SENCR knockdown disclose decreased expression of Myocardin and numerous smooth muscle contractile genes, whereas several promigratory genes are increased. Reverse transcription PCR and Western blotting experiments validate several differentially expressed genes after SENCR knockdown. Loss-of-function studies in scratch wound and Boyden chamber assays support SENCR as an inhibitor of smooth muscle cell migration.
SENCR is a new vascular cell-enriched, cytoplasmic lncRNA that seems to stabilize the smooth muscle cell contractile phenotype.

Download full-text


Available from: Joseph Miano, Feb 03, 2015
52 Reads
  • Source
    • "transcribedantisensefromthe5′endoftheFLI1geneand existsastwosplicevariants(Belletal.,2014).SENCRhasa cytoplasmiclocation(Belletal.,2014).RNA-seqexperiments inVSMCsafterSENCRknockdownrevealeddecreasedexpres- sionofmyocardinandnumeroussmoothmusclecontractile genes,whileseveralpromigratorygeneswereincreased.Loss-of- functionstudiesinscratchwoundandBoydenchamberassays suggestsSENCRasaninhibitorofVSMCmigration(Belletal., 2014). "
    [Show abstract] [Hide abstract]
    ABSTRACT: An aneurysm is a local dilatation of a vessel wall which is >50% its original diameter. Within the spectrum of cardiovascular diseases, aortic aneurysms are among the most challenging to treat. Most patients present acutely after aneurysm rupture or dissection from a previous asymptomatic condition and are managed by open surgical or endovascular repair. In addition, patients may harbor concurrent disease contraindicating surgical intervention. Collectively, these factors have driven the search for alternative methods of identifying, monitoring and treating aortic aneurisms using less invasive approaches. Non-coding RNA (ncRNAs) are emerging as new fundamental regulators of gene expression. The small microRNAs have opened the field of ncRNAs capturing the attention of basic and clinical scientists for their potential to become new therapeutic targets and clinical biomarkers for aortic aneurysm. More recently, long ncRNAs (lncRNAs) have started to be actively investigated, leading to first exciting reports, which further suggest their important and yet largely unexplored contribution to vascular physiology and disease. This review introduces the different ncRNA types and focus at ncRNA roles in aorta aneurysms. We discuss the potential of therapeutic interventions targeting ncRNAs and we describe the research models allowing for mechanistic studies and clinical translation attempts for controlling aneurysm progression. Furthermore, we discuss the potential role of microRNAs and lncRNAs as clinical biomarkers.
    Frontiers in Genetics 05/2015; 6(6). DOI:10.3389/fgene.2015.00125
  • Circulation Research 04/2014; 114(9):1366-8. DOI:10.1161/CIRCRESAHA.114.303896 · 11.02 Impact Factor
  • Arteriosclerosis Thrombosis and Vascular Biology 06/2014; 34(6):1124-5. DOI:10.1161/ATVBAHA.114.303504 · 6.00 Impact Factor
Show more