Alternative splicing in mechanically stretched podocytes as a model of glomerular hypertension

Abstract

BACKGROUND: Alterations in pre-mRNA splicing are crucial to the pathophysiology of various diseases. However, the effects of alternative splicing of mRNA on podocytes in hypertensive nephropathy are still unknown. The Sys_CARE project aimed to identify alternative splicing events involved in the development and progression of glomerular hypertension. METHODS: Murine podocytes were exposed to mechanical stretch, after which proteins and mRNA were analyzed by proteomics, RNA sequencing and several bioinformatic alternative splicing tools. RESULTS: Using transcriptomic and proteomic analysis, we identified significant changes in gene expression and protein abundance due to mechanical stretch. RNA-Seq identified over 3,000 alternative spliced genes after mechanical stretch, including all types of alternative splicing events. Among these, 17 genes exhibited an alternative splicing event across four different splicing analysis tools. From this group, we focused on Myl6, a component of the myosin protein complex, and Shroom3, an actin-binding protein essential for podocyte function. We identified two Shroom3 isoforms with significant expression changes under mechanical stretch, which was validated by qRT-PCR and in situ hybridization. Additionally, we observed an expression switch of two Myl6 isoforms after mechanical stretch, accompanied by an alteration in the C-terminal amino acid sequence. CONCLUSIONS: A comprehensive RNA-Seq analysis of mechanically stretched podocytes identified novel potential podocyte-specific biomarkers and highlighted significant alternative splicing events, notably in the mRNA of Shroom3 and Myl6.

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Titel	Alternative splicing in mechanically stretched podocytes as a model of glomerular hypertension
Medien	J. Am. Soc. Nephrol.
Verlag	Ovid Technologies (Wolters Kluwer Health)
Heft	9
Band	36
Verfasser	Francescapaola Mattias, Olga Tsoy, Elke Hammer, Alexander Gress, Prof. Dr. Stefan Simm, Chit Lio, Sabine Ameling, Kerstin Amann, Leonie Dreher, Ulrich Wenzel, Tim Kacprowski, Markus List, Olga Kalinina, Karlhans Endlich, Jan Baumbach, Uwe Völker, Nicole Endlich, Felix Kliewe
Seiten	1702–1715
Veröffentlichungsdatum	5.2025
Zitation	Mattias, Francescapaola; Tsoy, Olga; Hammer, Elke; Gress, Alexander; Simm, Stefan; Lio, Chit; Ameling, Sabine; Amann, Kerstin; Dreher, Leonie; Wenzel, Ulrich; Kacprowski, Tim; List, Markus; Kalinina, Olga; Endlich, Karlhans; Baumbach, Jan; Völker, Uwe; Endlich, Nicole; Kliewe, Felix (2025): Alternative splicing in mechanically stretched podocytes as a model of glomerular hypertension. J. Am. Soc. Nephrol. 36 (9), 1702–1715.