Kidney Week

Abstract: TH-PO876

Whole-Kidney 3D Imaging and Transcriptome Assessment in the UNx db/db Mouse Model of Diabetic Nephropathy

Session Information

• Diabetic Kidney Disease: Basic - I
  November 07, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Diabetic Kidney Disease

• 601 Diabetic Kidney Disease: Basic

Authors

• Sembach, Frederikke Emilie, Gubra, Hørsholm, Denmark

Frederikke Emilie Sembach,

• Boland, Brandon B., Gubra, Hørsholm, Denmark

Brandon B. Boland,

• Fink, Lisbeth N., Gubra, Hørsholm, Denmark

Lisbeth N. Fink,

• Roostalu, Urmas, Gubra, Hørsholm, Denmark

Urmas Roostalu,

• Skytte, Jacob Lercke, Gubra, Hørsholm, Denmark

Jacob Lercke Skytte,

• Secher, Thomas, Gubra, Hørsholm, Denmark

Thomas Secher,

• Fosgerau, Keld, Gubra, Hørsholm, Denmark

Keld Fosgerau,

• Vrang, Niels, Gubra, Hørsholm, Denmark

Niels Vrang,

• Jelsing, Jacob, Gubra, Hørsholm, Denmark

Jacob Jelsing,

• Hecksher-Sørensen, Jacob, Gubra, Hørsholm, Denmark

Jacob Hecksher-Sørensen,

• Pedersen, Tanja X., Gubra, Hørsholm, Denmark

Tanja X. Pedersen,

Background

Diabetic nephropathy (DN) is associated with increased cardiovascular risk and shortened survival. The DN therapeutic landscape has remained almost completely unchanged for decades, largely due to the lack of translatable preclinical models. In this study, we assessed renal changes in uninephrectomized (UNx) db/db mice using light sheet microscopy (LSM) and RNA sequencing to better define the structural and functional changes associated with early progression of DN.

Methods

UNx was performed in 7-8 week old male and female db/db mice. Sham-operated db/+ mice served as controls. Kidneys were preserved for histology, stereology, and RNA sequencing. In a separate study, UNx was performed in 18 week old male db/db mice. Mice were injected with lectin-594 immediately prior to termination to visualize glomerular morphology and to assess glomerular permeability via LSM. All mice were terminated at 24 weeks.

Results

Male and female UNx db/db mice showed similar progression of type 2 diabetes and urine albumin to creatinine ratio (UACR). Glomerular volume was increased in both genders relative to non-diabetic controls. Tubulointerstitial collagen III was increased in female UNx db/db vs. control mice, whereas glomerulosclerosis, as assessed by collagen IV/podocin colocalization, was significantly increased in male UNx db/db mice. Kidney RNAseq revealed increased expression of the glomerular marker Nphs1 and the tubule markers Lcn2, Spp1, Havcr1, confirming glomerular hypertrophy and kidney injury. Glomerular number was determined by LSM and subsequent stereological assessment on the same kidneys, allowing direct comparison of the two methodologies. UNx and control mice demonstrated similar glomerular numbers (~16,000), whereas UNx mice had increased glomerular volume (81,000 mm³ vs 145,000 mm³). Moreover, LSM revealed a marked increase in cortical lectin-594 staining, indicating glomerular leakage and emerging glomerular dysfunction in UNx db/db mice.

Conclusion

The use of whole-kidney 3D imaging in tandem with RNAseq offers a novel means of assessing individual glomerular structure and function during DN progression and represents a powerful technique to profile the preclinical efficacy of forthcoming DN therapeutics.

Funding

• Commercial Support –