Abstract: TH-PO0274
Renal Neuropeptide FF Receptor 2, via the Stress Response Element Within Its Gene Promoter Region and Interaction with Dopamine D1 Receptor and Angiotensin II Type 1 Receptor, Regulates Blood Pressure
Session Information
- Hypertension and CVD: Mechanisms
November 06, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Hypertension and CVD
- 1601 Hypertension and CVD: Basic
Authors
- Qaddumi, Waleed N., The George Washington University School of Medicine and Health Sciences, Washington, District of Columbia, United States
- Amatya, Bibhas, The George Washington University School of Medicine and Health Sciences, Washington, District of Columbia, United States
- Villar, Van Anthony M., The George Washington University School of Medicine and Health Sciences, Washington, District of Columbia, United States
- Asico, Laureano D., The George Washington University School of Medicine and Health Sciences, Washington, District of Columbia, United States
- Rozyyev, Selim, The George Washington University School of Medicine and Health Sciences, Washington, District of Columbia, United States
- Feranil, Jun, The George Washington University School of Medicine and Health Sciences, Washington, District of Columbia, United States
- Armando, Ines, The George Washington University School of Medicine and Health Sciences, Washington, District of Columbia, United States
- Felder, Robin Allen, University of Virginia School of Medicine, Charlottesville, Virginia, United States
- Jose, Pedro A., The George Washington University School of Medicine and Health Sciences, Washington, District of Columbia, United States
- Lee, Hewang, The George Washington University School of Medicine and Health Sciences, Washington, District of Columbia, United States
Background
Neuropeptide FF (NPFF) receptor 2 (NPFFR2) is a receptor with diverse biological function. Renal-restricted stimulation, via subcapsular infusion, of NPFFR2 and angiotensin II (Ang II) type 1 receptor (AT1R) increases blood pressure (BP) whereas the dopamine D1 receptor (D1R) decreases BP.
Methods
The Stress Response Element (SRE) within the NPFFR2 promoter was analyzed by in silico and confirmed by luciferase assay. Colocalization and protein-protein interaction were determined by confocal microscopy and co-immunoprecipitation respectively. Effect on Na+ transport in renal proximal tubule cells (RPTCs) was determined by fluorescence lifetime imaging (FLIM). BP was measured from the carotid artery in conscious (telemetry) and anesthetized mice.
Results
In silico analysis revealed an SRE within the NPFFR2 promoter. Luciferase assay showed that wild-type promoter activity of NPFFR2 gene was increased (4.1±0.05-fold, n=3) by low NaCl (90 mM) but was prevented by the mutant NPFFR2. The renal subcapsular infusion of specific SRE antigene RNA abrogated the increase in NPFFR2 protein expression and systolic BP (83.3±1.3 vs 65±0.6 mm Hg, P<0.05) in C57BL/6J mice fed a low salt diet for 3 days. In human RPTCs, NPFFR2 colocalized and co-immunoprecipitated with D1R. Fenoldopam (Fen, 1 µM), a D1R agonist, impaired Na+ transport from inside to outside the cell, shown by increased intracellular Na+ concentration, detected by the increase in lifetime t2 measured by FLIM. The Fen-mediated increase in lifetime was attenuated by NPFF (100 nM), indicating an antagonistic interaction between NPFFR2 and D1R on Na+ transport. In kidneys, NPFFR2 also colocalized with AT1R, and NPFFR2 co-immunoprecipitated with AT1R. The renal subcapsular infusion of NPFF (10 µg in 100 µL) for 7 days increased the systolic BP, which was attenuated by the femoral vein infusion of Fen (2 µg/kg/min) by osmotic minipump, whereas, systolic BP was individually and synergistically increased by renal subcapsular infusion of NPFF (10 µg/100 µL) and Ang II (20 µg/100 µL) 0.5 µL/hr for 7 days.
Conclusion
Renal NPFFR2 regulates BP through SRE within its promotor and its interaction with D1R and AT1R in the kidney.
Funding
- NIDDK Support