Abstract: TH-PO755
The Role of miR-143-3p in Podocyte Damage in Preeclampsia
Session Information
- Sex and Reproductive Factors in Kidney Health
November 03, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Women's Health and Kidney Diseases
- 2100 Women's Health and Kidney Diseases
Authors
- Suvakov, Sonja, Mayo Clinic Division of Nephrology and Hypertension, Rochester, Minnesota, United States
- Gojkovic, Tamara, Mayo Clinic Division of Nephrology and Hypertension, Rochester, Minnesota, United States
- Garovic, Vesna D., Mayo Clinic Division of Nephrology and Hypertension, Rochester, Minnesota, United States
Background
Preeclampsia (PE) is a pregnancy-specific disease characterized by hypertension accompanied by either proteinuria and/or end-organ damage. Previous research has shown that PE is associated with podocyturia and an increased number of podocyte-derived urinary extracellular vesicles (EV), but underlying the mechanism is not well understood. We aimed to elucidate the potential role of miR-143-3p in podocyte injury in PE.
Methods
XRNA Exosome RNA-Seq Library Kit and qPCR were used to identify miRNA candidate(s) to study. Micro RNA 143-3p was identified and its expression was studied in an immortalized human podocyte cell line treated with either PE sera or TNF-α. We also investigated miR143-3p roles in inflammation, VEGF secretion, podocyte migration, both in the absence and presence of its inhibitor.
Results
Upon assessing differential miRNA expressions in urinary EV and validating candidates relevant for kidney injury, we identified miR143-3p as significantly increased in PE. Treatment of an immortalized podocyte cell line with either pooled PE sera or TNF-α. Treatment with pooled sera showed increase in miR143-3p expression in dose-dependent manner. In conditioned media, TGF-β was increased while VEGF was decreased, both reversed by treatment with mir-143-3p inhibitor. Podocytes incubated with TNF-α showed higher gene expressions of miR143-3p and pro-inflammatory cytokines, IL-6, IL-8 and MCP-1, which were attenuated by a miRNA-143-3p inhibitor. No significant changes in p16 and p21 gene expressions were observed with either treatment. Beta-gal staining for senescence indicated increased senescent phenotype in podocytes treated with PE sera. Wound healing assay showed that both treatments decreased migration of podocytes, whereas the miRNA inhibitor improved migration. Finally, the total number of both podocin and nephrin positive EVs was increased in cultured media upon podocyte treatment with PE sera.
Conclusion
Micro RNA 143-3p is abundant in urinary EVs of preeclamptic women. It mediates expressions of pro-inflammatory cytokines in podocytes and affects their migration. These results indicate that miR-143-3p may contribute to the pathophysiology of podocyte injury in preeclampsia.