Publication

EDN1-AS, A Novel Long Non-coding RNA Regulating Endothelin-1 in Human Proximal Tubule Cells

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  • 06/17/2025
Type of Material
Authors
    Lauren G. Douma, University of FloridaKristen Solocinski, University of FloridaSarah H. Masten, University of FloridaDominique H. Barral, University of FloridaSarah J. Barilovits, University of FloridaLauren Jeffers, Emory UniversityKareme D. Alder, Yale UniversityRavi Patel, University of FloridaCharles S. Wingo, University of FloridaKevin D. Brown, University of FloridaBrian D. Cain, University of FloridaMichelle L. Gumz, University of Florida
Language
  • English
Date
  • 2020-03-13
Publisher
  • Frontiers
Publication Version
Copyright Statement
  • © 2020 Douma, Solocinski, Masten, Barral, Barilovits, Jeffers, Alder, Patel, Wingo, Brown, Cain and Gumz.
License
Final Published Version (URL)
Title of Journal or Parent Work
Volume
  • 11
Start Page
  • 209
Grant/Funding Information
  • This work was supported by the National Institutes of Health (NIH) R21AG052861 and 1R01DK109570-01A1 (MG), American Heart Association Grant-in-Aid (16GRNT31220009) (MG), University of Florida Department of Medicine Gatorade Trust (MG), American Heart Association Postdoctoral Fellowship Grants 18POST34030210 (LD), and NIH Grant T32-DK-104721 awarded to the University of Florida (UF) Division of Nephrology (LD).
Abstract
  • Endothelin-1 (ET-1) is a peptide hormone that functions as a vasoconstrictor in the vasculature, whereas in the collecting duct of the kidney it exerts blood pressure-lowering effects via natriuretic actions. Aberrant ET-1 signaling is associated with several pathological states including hypertension and chronic kidney disease. ET-1 expression is regulated largely through transcriptional control of the gene that encodes ET-1, EDN1. Here we report a long, non-coding RNA (lncRNA) that appears to be antisense to the EDN1 gene, called EDN1-AS. Because EDN1-AS represents a potential novel mechanism to regulate ET-1 expression, we examined the regulation of EDN1-AS expression and action. A putative glucocorticoid receptor response (GR) element upstream of the predicted EDN1-AS transcription start site was identified using the ENCODE database and the UCSC genome browser. Two homozygous deletion clones of the element were generated using CRISPR/Cas9. This deletion resulted in a significant increase in the expression of EDN1-AS, which was associated with increased secretion of ET-1 peptide from HK-2 cells (two-fold increase in KO cells vs. CNTL, n = 7, P < 0.05). Phenotypic characterization of these CRISPR clones revealed a difference in cell growth rates. Using a standard growth assay, we determined that the KO1 clone exhibited a three-fold increase in growth over 8 days compared to control cells (n = 4, P < 0.01) and the KO2 clone exhibited a two-fold increase (n = 4, P < 0.01). These results support a role for EDN1-AS as a novel regulatory mechanism of ET-1 expression and cellular proliferation.
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Keywords
Research Categories
  • Biology, Genetics
  • Health Sciences, Medicine and Surgery

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