Abstract: PUB108
Spatial-omic Profiling of Human Kidney Tissues Using SeqStain Stratifies Disease Pathology
Session Information
Category: Diabetic Kidney Disease
- 701 Diabetic Kidney Disease: Basic
Authors
- Vattikota, Anirudh, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States
- Youssef, Mohamed, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States
- Gurusamy Kamaraj, Sowmya, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States
- Venkatesh, Ishwarya, Rush University Medical Center, Chicago, Illinois, United States
- Shaw, Ameera M., Rush University Medical Center, Chicago, Illinois, United States
- Cimbaluk, David J., Rush University Medical Center, Chicago, Illinois, United States
- Gupta, Vineet, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States
Group or Team Name
- Internal Medicine.
Background
A deep understanding of the pathophysiology of the tissue microenvironment is crucial for scientific advancements. Newer multiplex imaging-based methods are providing important details about the histoarchitecture of tissue. We previously developed a novel user-friendly tissue imaging method, called SeqStain, allows rapid immunofluorescence based multiplexed tissue imaging and analyses. Here, we utilize this approach to understand the complex spatial organization of cells and their relationships with each other with both healthy subjects, and patients with diabetic nephropathy and lupus nephritis.
Methods
We generated antibodies conjugated with fluorescently-labelled-DNA oligonucleotides for analyzing multiple kidney-specific antigens using sequential cycles of staining and de-staining on each tissue section. We probed different areas of histologically 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 commercially available reagents and a confocal microscope. This allowed us to precisely image up to 20 antigens on single tissue specimens for healthy subjects, as well as 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 is a versatile, gentle, and easy to use method for multiplex imaging making it highly effective in mapping the spatial relationship of cells within the kidney. The spatial relationship maps generated by this method will offer valuable new insights into disease pathophysiology and enhance future diagnostics and treatments for LN and DN.
Funding
- Private Foundation Support