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

Area of Science:

• Molecular Biology
• Genetics
• Cardiovascular Physiology

Background:

• Endothelin-1 (ET-1) is a peptide hormone with dual roles: vasoconstriction in blood vessels and blood pressure lowering via natriuresis in the kidney.
• Dysregulated ET-1 signaling is implicated in hypertension and chronic kidney disease.
• ET-1 gene (EDN1) expression is primarily controlled transcriptionally.

Purpose of the Study:

• To investigate the role of a newly identified antisense long non-coding RNA (lncRNA), EDN1-AS, in regulating ET-1 expression.
• To explore the regulatory mechanisms controlling EDN1-AS expression and its functional consequences.

Main Methods:

• Bioinformatic analysis identified a putative glucocorticoid receptor (GR) response element upstream of the EDN1-AS transcription start site.
• CRISPR/Cas9 gene editing was used to generate homozygous deletion clones (KO) of this GR element.
• ET-1 peptide secretion and cell proliferation rates were measured in control (CNTL) and KO cells (HK-2).

Main Results:

• Deletion of the GR element significantly increased EDN1-AS expression.
• Increased EDN1-AS expression correlated with a two-fold increase in ET-1 peptide secretion from HK-2 cells.
• CRISPR clones exhibited altered cell growth rates, with KO1 and KO2 showing 3-fold and 2-fold increases in proliferation, respectively.

Conclusions:

• EDN1-AS acts as a novel regulator of ET-1 expression.
• EDN1-AS influences cellular proliferation, suggesting a role in disease pathogenesis.
• These findings highlight EDN1-AS as a potential therapeutic target for conditions involving aberrant ET-1 signaling.