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Kidney Week

Abstract: FR-OR19

Advanced Light Sheet Microscopy and 3D Image Analyses of Kidney Injury, Glomerulosclerosis, and Fibrosis in a Mouse Model of Diabetic Kidney Disease

Session Information

Category: Diabetic Kidney Disease

  • 601 Diabetic Kidney Disease: Basic

Authors

  • Østergaard, Mette Viberg, Gubra Aps, Horsholm, Denmark
  • Roostalu, Urmas, Gubra Aps, Horsholm, Denmark
  • Bak, Stine Thorhauge, Gubra Aps, Horsholm, Denmark
  • Skytte, Jacob Lercke, Gubra Aps, Horsholm, Denmark
  • Vrang, Niels, Gubra Aps, Horsholm, Denmark
  • Hecksher-Sørensen, Jacob, Gubra Aps, Horsholm, Denmark
Background

Development of novel therapies for diabetic kidney disease (DKD) and other glomerulopathies is challenged by poor translatability of preclinical animal models. Novel biomarkers are sought to close this translational gap. Using 3D imaging techniques and advanced image analyses, we aimed to develop a method for quantification of kidney injury and fibrosis in a preclinical mouse model of progressive DKD.

Methods

Kidneys from hypertensive uninephrectomized db/db mice (reninAAV UNx db/db) and healthy controls were fixed and processed for whole-mount immunohistochemistry and light sheet microscopy (LSM) to assess and quantify tubular injury by KIM-1, and fibrosis and glomerulosclerosis by tenascin in the intact kidney. Using 3D image analysis, the distribution and intensity of KIM-1 and tenascin were determined. To correlate 3D imaging endpoints with DKD severity, kidney fibrosis and injury was characterized using standard methodologies including 2D histology.

Results

In reninAAV UNx db/db mice, tenascin was present in glomeruli as showed by its overlap with podocin. A sub-population of glomeruli with augmented tenascin intensity, but with no overlap of podocin was identified indicating that these glomeruli have global glomerulosclerosis and loss of podocytes. Tubulointerstitial tenascin was limited. These findings correlated with traditional histopathological assessment of glomerulosclerosis scoring and fibrosis quantification in PAS and collagen 3 stained kidney section, respectively. KIM-1 positive tubuli were also visualized in intact kidneys from reninAAV UNx db/db mice and showed a heterogenous pattern across the kidney. KIM-1 was clearly localized to the proximal tubules and was also present in parietal cells in a subpopulation of glomeruli. These observations correlated with 2D IHC stains of KIM-1. Kidneys from healthy controls were KIM-1 negative in both 3D and 2D.

Conclusion

Development of advanced microscopy and 3D imaging technologies allows for assessment of kidney fibrosis and injury in the intact mouse kidney. Thereby, this 3D imaging technique can be used to support functional and 2D histological readouts in mouse models to improve their translatability in the study of disease mechanisms and drug discovery for DKD.