Kidney Week

Abstract: FR-OR117

Assessing Glomerular Cellularity in Diabetic Human Kidney Biopsies with 3D Tissue Cytometry: Implications for Disease Progression

Session Information

• Translating Discovery to Patients with Diabetic Kidney Disease
  November 08, 2019 | Location: 207, Walter E. Washington Convention Center
  Abstract Time: 05:42 PM - 05:54 PM

Category: Diabetic Kidney Disease

• 601 Diabetic Kidney Disease: Basic

Authors

• Sabo, Angela R., Indiana University School of Medicine, Indianapolis, Indiana, United States

Angela R. Sabo

• Ferkowicz, Michael J., Indiana University, Indianapolis, Indiana, United States

Michael J. Ferkowicz,

• Eadon, Michael T., Indiana University Division of Nephrology, Indianapolis, Indiana, United States

Michael T. Eadon,

• Phillips, Carrie L., Indiana University School of Medicine, Indianapolis, Indiana, United States

Carrie L. Phillips,

• Kelly, Katherine J., Indiana University, Indianapolis, Indiana, United States

Katherine J. Kelly,

• Sutton, Timothy A., Indiana University School of Medicine, Indianapolis, Indiana, United States

Timothy A. Sutton,

• Dagher, Pierre C., Indiana University, Indianapolis, Indiana, United States

Pierre C. Dagher,

• Winfree, Seth, Indiana University School of Medicine, Indianapolis, Indiana, United States

Seth Winfree,

• El-Achkar, Tarek M., Indiana University, Indianapolis, Indiana, United States

Tarek M. El-Achkar,

Background

Diabetic nephropathy (DN) is a leading cause of kidney disease worldwide. At the tissue level, DN has been classified by morphology including glomerular size and cellularity. Large scale 3D multi-fluorescence imaging and 3D tissue cytometry (3DTC) provides a unique and efficient way to quantitate and characterize the cellularity of glomeruli. Here, we examined the changes in cellular density across multiple kidney biopsy specimens with DN, compared to reference nephrectomy tissue. We also sought to determine whether the changes in cellularity are indicative of a focal or diffuse response.

Methods

5 non-DN reference nephrectomy specimens and 5 kidney biopsy specimens with DN were stained with 4 to 8 markers and imaged by confocal fluorescence microscopy. Glomeruli were isolated digitally and analyzed using 3DTC with Volumetric Tissue Exploration and Analysis (VTEA) software. .Cell density and immune cell subtypes were determined for each glomerulus within each specimen.

Results

When comparing all glomeruli from diabetics to those from the reference specimens, the average cellular density was increased in the diabetic group: 592715 ± 116469 vs. 259556 ± 8107 cells/mm³, respectively (p < 0.05). When comparing between individual specimen, there were no significant differences seen in glomerular cellular density across reference tissues, but three of the five diabetic specimens showed significant increases in cellularity, albeit to different levels (p<0.05 using one-way ANOVA compared to reference). Interestingly, within each biopsy with DN, the cellular densities in all glomeruli were comparable. When examining cell subtype, the contribution of immune cells to the increased cellularity in diabetes was minimal.

Conclusion

3DTC using VTEA is a powerful and efficient tool to assess glomerular cellularity in biopsy specimens. Using this tool, we detected an increase in glomerular cell density in DN. Importantly, our data suggest that diabetes uniformly alters the cellularity across all glomeruli within a biopsy specimen, rather than a selective effect on a subset of glomeruli. These findings increase our understanding of the dynamics of the progression of diabetic kidney disease and may aid pathologists' interpretation of specimens.

Funding

• NIDDK Support