Abstract: SA-OR055
Stratifying Diabetic Kidney Disease Through Spatialomic Analysis with SeqStain
Session Information
- Kidney Disease with Diabetes: Translational Science Breakthroughs
November 08, 2025 | Location: Room 372A, Convention Center
Abstract Time: 04:30 PM - 04:40 PM
Category: Diabetic Kidney Disease
- 701 Diabetic Kidney Disease: Basic
Authors
- Gurusamy Kamaraj, Sowmya, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States
- Youssef, Mohamed A., The University of Texas Medical Branch at Galveston, Galveston, Texas, United States
- Vattikota, Anirudh, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States
- Venkatesh, Ishwarya, Rush University, Chicago, Illinois, United States
- Shaw, Ameera M., Rush University, Chicago, Illinois, United States
- Cimbaluk, David J., Rush University, Chicago, Illinois, United States
- Gupta, Vineet, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States
Background
The human kidney contains over 20 distinct cell types, each with essential functions and organized into spatially defined subpopulations. Diabetic kidney disease (DKD) develops in 40% of people with type 2 diabetes mellitus and is the critical cause of end-stage kidney disease worldwide. Diabetic kidney disease (DKD) is a complex, multifactorial condition characterized by significant variability in both its underlying pathophysiology and individual responses to treatment, shaped by genetic and environmental factors. Diagnosing and staging DKD through routine measurements of standard laboratory markers lacks the resolution to capture the complex and varied mechanisms underlying disease progression in individual patients. Towards this goal, we demonstrate using Seqstain, a previously developed novel user-friendly tissue imaging technique that allows rapid immunofluorescence-based multiplexed tissue imaging and analysis. We utilize this approach to understand the complex spatial organization of cells and their relationships with each other, using healthy subjects and patients with DKD.
Methods
We used SeqStain, a sequential immunofluorescence method with DNA-labeled antibodies, to profile multiple kidney-specific antigens. Histologically relevant regions were probed through cycles of staining, imaging, and label removal, enabling high-plex spatial mapping via standard fluorescence microscopy and image analysis software.
Results
We generated multiplexed images from frozen kidney sections of healthy individuals and DKD patients. Using confocal microscopy and commercial reagents, we imaged up to 20 antigens per section, covering structures like glomeruli, proximal tubules, and podocytes. We analyzed spatial cell organization and diabetes-related changes in cellular neighborhoods. SeqStain was used to identify correlations and anti-correlations among protein markers around each segmented cell.
Conclusion
SeqStain multiplex imaging is a gentle, user-friendly method for mapping cellular spatial relationships in the kidney. It enables sensitive, selective profiling of cell populations and reveals key features of renal structure and function. The resulting spatial maps offer insights into DKD pathophysiology and may guide future diagnostics and therapies.
Funding
- NIDDK Support