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Kidney Week

Abstract: FR-PO688

Spatialomic Profiling of Human Kidney Tissues Stratifies Disease Pathology

Session Information

Category: Glomerular Diseases

  • 1401 Glomerular Diseases: From Inflammation to Fibrosis

Authors

  • Venkatesh, Ishwarya, Rush University Medical Center, Chicago, Illinois, United States
  • Shaw, Ameera M., Rush University Medical Center, Chicago, Illinois, United States
  • Gupta, Vineet, Rush University Medical Center, Chicago, Illinois, United States
  • Cimbaluk, David J., Rush University Medical Center, Chicago, Illinois, United States
Background

To make any scientific advancements, there should be an in-depth understanding of pathophysiology of tissue microenvironment. Newer multiplex imaging-based methods are providing important insights into cytoarchitecture of tissue. We recently developed a novel tissue imaging method, termed SeqStain, that allows rapid and easy immunofluorescence based multiplexed tissue imaging and analyses. Here, we utilize this approach to understand the complex assemblage of cells and their correlations with each other in both healthy subjects and patients with various glomerular diseases.

Methods

We synthesized fluorescently-DNA-tagged antibodies for analyzing multiple kidney-specific antigens using cycles of staining and de-staining on a single tissue section. We probed different histological regions relevant to the kidney and used conventional fluorescence microscopy for imaging the tissues and HALO software for image analyses.

Results

We analyzed both paraffin-fixed and frozen tissue sections using off-the-shelf reagents and a confocal microscope. We were able to accurately image tens of antigens on single tissue specimens for healthy subjects, and from patients with lupus nephritis (LN) and diabetic nephropathy (DN). Analysis of various cellular biomarkers indicated enrichment of specific cellular clusters into distinct neighborhoods.

Conclusion

SeqStain proved to be a versatile, gentle, and easily adaptable method for multiplex imaging that can be highly effective in obtaining a spatial map of kidney. Generated spatial maps will provide important new insights about the disease pathobiology and improve future diagnostics and therapeutics for LN and DN.