Abstract: SA-PO0478
miRNA Expression of Urinary Extracellular Vesicles in Patients with Gitelman Syndrome: The Role of Hsa-let-7d-3p
Session Information
- Fluid, Electrolyte, and Acid-Base Disorders: Basic Research
November 08, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Fluid, Electrolytes, and Acid-Base Disorders
- 1101 Fluid, Electrolyte, and Acid-Base Disorders: Basic
Authors
- Sung, Chih-Chien, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan
- Tseng, Min-hua, Department of Pediatrics, Chang Gung Memorial Hospital, Taoyuan, Taiwan
- Lin, Shih-Hua P., Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan
Background
Gitelman syndrome (GS) is an inheritable renal tubule disorder caused by the SLC12A3 mutation, leading to the sodium chloride cotransporter (NCC) defects. The role of miRNAs in regulating renal transporters in GS remains unclear. Our aim is to identify miRNA expression from urinary extracellular vesicles (uEVs) in GS patients and explore their role in renal tubular regulation.
Methods
Both uEVs from 23 genetically confirmed GS patients and renal biopsied tissue from another 3 GS patients were extracted for small RNA sequencing. Real-time PCR of uEVs (n = 11), immunofluorescence staining of Nedd4L in NccS707X/S707X knock-in mice (GS animal model), and dual luciferase reporter assays were performed for validation.
Results
Small RNA sequencing identified 338 miRNAs from uEVs and 652 miRNAs from renal biopsied tissues. Among the differentially expressed miRNAs, 20 were upregulated and 23 downregulated in uEVs, while renal biopsied tissues showed 30 upregulated and 23 downregulated miRNAs. Four miRNAs (hsa-let-7d-3p, hsa-miR-362-5p, hsa-miR-30c-5p, and hsa-miR-30b-5p) overlapped with both uEVs and renal tissues. Notably, the Gene Ontology Biological Process terms of the distinct upregulated hsa-let-7d-3p target genes were related to “ion transport” and “membrane depolarization”, especially in NEDD4L. Real-time PCR of uEVs from another 11 GS patients confirmed significantly elevated hsa-let-7d-3p compared to healthy controls. Decrease of Nedd4l expression in collecting duct was also confirmed in NccS707X/S707X knock-in mice. Dual luciferase assays further demonstrated hsa-let-7d-3p negatively regulated the expression of NEDD4L.
Conclusion
The miRNA expression could be isolated from uEVs and their differentially expressed miRNAs, especially has-let-7d-3p, may play critical role in renal tubular transporter regulation, offering novel insights into the regulation of tubular transporters in GS.
Funding
- Government Support – Non-U.S.