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Abstract: SA-PO0752

Wt1os lncRNA as a Novel Regulator of Podocyte Function and Glomerular Pathophysiology

Session Information

Category: Glomerular Diseases

  • 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology

Authors

  • Mueller, Roman-Ulrich, Universitatsklinikum Koln Klinik II fur Innere Medizin Nephrologie Rheumatologie Diabetologie und Allgemeine Innere Medizin, Cologne, NRW, Germany
  • Filipów, Samantha, Universitatsklinikum Koln Klinik II fur Innere Medizin Nephrologie Rheumatologie Diabetologie und Allgemeine Innere Medizin, Cologne, NRW, Germany
  • Antczak, Philipp, Universitatsklinikum Koln Klinik II fur Innere Medizin Nephrologie Rheumatologie Diabetologie und Allgemeine Innere Medizin, Cologne, NRW, Germany
  • Ignarski, Michael, Universitatsklinikum Koln Klinik II fur Innere Medizin Nephrologie Rheumatologie Diabetologie und Allgemeine Innere Medizin, Cologne, NRW, Germany
Background

Long non-coding RNAs (lncRNAs) are key regulators of gene expression, influencing processes such as chromatin remodeling, transcription, and translation. While their dysregulation has been linked to various diseases, including kidney disorders, their specific roles in glomerular biology remain poorly understood. Previously, we found that Wt1os, a conserved lncRNA transcribed divergently from the essential podocyte gene Wt1, was differentially regulated in several mouse models of podocyte disease. Consequently, we set out, to study the role of Wt1os in podocyte biology and disease.

Methods

A Wt1os loss of function mouse line (Wt1os-/-) was generated using CRISPR/Cas9. To investigate transcriptional changes, we performed RNAseq of glomeruli isolated from Wt1os-/- and control mice. STED imaging of optically cleared kidneys was used to visualize podocyte foot processes. Urinary albumin and creatinine levels were quantified via ELISA and biochemical assays. In cultured podocytes, CRISPRa was used for Wt1os overexpression, while siRNA or GapmeRs were used for knockdown. RNAscope was employed to determine the subcellular localization of Wt1os/WT1-AS. Cell death was assessed using the Incucyte system and calcium dynamics were measured by Fluo-4 AM-based imaging.

Results

Loss of Wt1os in mice led to podocyte foot process effacement, visualized by STED imaging, and albuminuria. RNAseq of isolated glomeruli revealed exon skipping in Wt1os-/- mice and mild but significant transcriptomic changes in the glomerulus. RNAScope showed Wt1os as well as its human orthologue to be nuclear lncRNAs. Notably, Rgs1 was consistently downregulated following Wt1os/WT1-AS loss in both the mouse and cultured podocytes models, suggesting its role as a downstream effector. Interestingly, Wt1os promoted calcium mobilization in an Rgs1-dependent manner in cultured murine podocytes. In cultured human podocytes, knockdown of WT1-AS impaired cell viability.

Conclusion

Our findings identify Wt1os as a novel lncRNA regulator of podocyte biology, acting through Rgs1-dependent calcium signaling. This study expands the current understanding of lncRNA function in glomerular disease and highlights Wt1os as a potential therapeutic target in FSGS.

Funding

  • Government Support – Non-U.S.

Digital Object Identifier (DOI)