Abstract: SA-PO769
Regulation of Edn1 by Its Antisense Long Non-Coding RNA, Edn1-AS
Session Information
- Hypertension and CVD: Mechanisms
November 05, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Hypertension and CVD
- 1503 Hypertension and CVD: Mechanisms
Authors
- Juffre, Alexandria, University of Florida, Gainesville, Florida, United States
- Douma, Lauren G., University of Florida, Gainesville, Florida, United States
- Wingo, Charles S., University of Florida, Gainesville, Florida, United States
- Gumz, Michelle L., University of Florida, Gainesville, Florida, United States
Group or Team Name
- Gumz Laboratory
Background
The peptide hormone endothelin-1 (ET-1) decreases renal sodium reabsorption by inhibiting epithelial sodium channel (ENaC) activity. Dysfunctional regulation of sodium handling can cause hypertension and ET-1 is a therapeutic target. We discovered a long non-coding RNA (lncRNA), Edn1-AS, which is antisense to the Edn1 mRNA. Using CRISPR-modified human proximal tubule cells (HK-2), we previously found that cis over-expression of Edn1-AS (expressed from the same locus as Edn1) resulted in increased ET-1 levels in cultured human proximal tubule cells (HK-2). The goal of this study was to test the effect of trans-overexpression of Edn1-AS (from an exogenous DNA construct) on Edn1 expression. Based on our previous results, we hypothesized that trans over-expressing Edn1-AS in collecting duct cells would increase ET-1 mRNA expression.
Methods
Inducible Edn1-AS overexpressing IMCD-3 and mpkCCD cells were generated using Tet-ONE systems (Takara Clontech), which increases Edn1-AS expression after treatment with doxycycline. Edn1-AS lncRNA and Edn1 mRNA levels were determined by strand-specific RT-PCR or qPCR.
Results
Similarly to cis over-expression in HK-2 cells, trans over-expression of Edn1-AS in IMCD-3 and mpkCCD cells showed increased Edn1 levels (n=3, p < 0.05 by unpaired t-test).
Conclusion
Consistent with our previous results in HK-2 cells, increased Edn1 expression was observed in Edn1-AS overexpressing IMCD-3 and mpkCCD cells. These data suggest that Edn1-AS is a positive regulator of Edn1, regardless of whether Edn1-AS is over-expressed in a cis or trans manner. Subsequent studies will seek to determine the mechanism of action of Edn1-AS on Edn1 expression which may involve chromatin remodeling. Future investigation of Edn1-AS's regulation of Edn1 could lead to the identification of novel therapeutic targets for treatment of hypertension.
Funding
- NIDDK Support