Abstract: TH-PO550
High-Resolution Morphometry and Spatial Transcriptomics to Reveal Cellular and Molecular Mechanisms of Podocyte Effacement
Session Information
- Glomerular Diseases: Omics, Biomarkers, and Tools
October 24, 2024 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology
Authors
- Bohnenpoll, Tobias, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Skroblin, Philipp, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Pospiech, Johannes, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Guhl, Anna, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Liebe, Nils, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Lomow, Alexander, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Wiechers, Carolin, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Seip, Britta, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Zheng, Menglin, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Helwig, Michael, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Fritsch, Rüdiger, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Radresa, Olivier, Evotec International GmbH, Göttingen, Niedersachsen, Germany
- Kramer, Kristina, Abberior Instruments GmbH, Göttingen, Niedersachsen, Germany
- Grimm, Florian, Abberior Instruments GmbH, Göttingen, Niedersachsen, Germany
- Wurm, Christian A., Abberior Instruments GmbH, Göttingen, Niedersachsen, Germany
- Donnert, Gerald, Abberior Instruments GmbH, Göttingen, Niedersachsen, Germany
- Andag, Uwe, Evotec International GmbH, Göttingen, Niedersachsen, Germany
Background
Podocyte injury and effacement precede proteinuria and result in characteristic glomerular lesions associated with focal segmental glomerulosclerosis (FSGS) and other glomerulopathies. These lesions have long been considered distinct disease entities, but their cellular and molecular features remain poorly characterized. Stimulated emission depletion (STED) is a light microscopy technique which provides a unique opportunity to assess podocyte foot process morphology in unprecedented detail. Here, we combine super-resolution STED morphometry and spatial transcriptomics to unravel the cellular and molecular mechanisms underlying podocyte injury and effacement in kidney biopsies from the NURTuRE patient cohort.
Methods
Adjacent thin sections of formalin-fixed, paraffin-embedded biopsies from 18 NURTuRE patients diagnosed with FSGS and other glomerulopathies and 2 healthy living donors were analyzed using STED microscopy (Facility line, Abberior) and spatial transcriptomics (Visium HD, 10x Genomics). Podocyte foot processes and filtration slits were stained for synaptopodin and nephrin, and super-resolution scans were exported for morphometry.
Results
Clinical-grade biopsies were selected to represent patients with early, intermediate, and end-stage disease, maximizing the range of kidney function and proteinuria. High-resolution spatial transcriptomics allowed identification of tissue niches based on the expression of cell type-specific and mechanistic signatures, representing distinct glomerular and tubulointerstitial microenvironments with different cellular composition. Unbiased clustering of spatially and molecularly defined niches stratified glomeruli along the continuum of podocyte injury and glomerular remodeling. Interestingly, glomerular niches were associated with different slit diaphragm lengths and densities, as determined by podocyte STED morphometry, depending on their position along the disease continuum.
Conclusion
Further integration of molecular, morphometric and clinical data has the potential to reveal cellular and molecular mechanisms of podocyte injury and glomerular remodeling and will aid in mechanistic disease understanding.
Funding
- Commercial Support – Evotec International GmbH, Abberior GmbH