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Abstract: TH-PO588

SeqStain Based Spatialomic Profiling of Human Kidney Tissues Identifies Cellular Neighborhoods

Session Information

  • Pathology and Lab Medicine
    November 03, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
    Abstract Time: 10:00 AM - 12:00 PM

Category: Glomerular Diseases

  • 1302 Glomerular Diseases: Immunology and Inflammation

Authors

  • Venkatesh, Ishwarya, Rush University Medical Center, Chicago, Illinois, United States
  • Varma, Disha, Rush University Medical Center, Chicago, Illinois, United States
  • Koh, Kwi Hye, Rush University Medical Center, Chicago, Illinois, United States
  • Nguyen, Billy, 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

Group or Team Name

  • Team SeqStain
Background

An improved understanding of the underlying cellular heterogeneity of human kidney tissue is essential for providing an accurate disease diagnosis, rate of progression and potential therapeutic avenues for a variety of chronic kidney disease pathologies. Newer multiplex imaging based methods are providing such tools for implementation in the clinical setting in the future. We recently developed a novel tissue imaging method, termed SeqStain, for immunofluorescence based multiplexed tissue imaging and analyses. Here, we describe utility of this approach for improving our understanding of kidney tissue samples from healthy subjects and patients with various glomerular diseases.

Methods

SeqStain utilizes fluorescent DNA-tagged antibodies and antibody-fragments for analyzing tens of kidney-specific analytes in a single tissue section. We designed and optimized SeqStain multiplex panels with sets of antibodies to probe different histological regions relevant to the kidney and used conventional fluorescence microscopy set-up for imaging and analyses.

Results

We show that SeqStain is an efficient method for multiplex imaging of both paraffin-fixed and frozen tissue sections. Image acquisition using off-the-shelf components, and confocal microscopes that are widely available in laboratories, we were able to accurately image tens of antigens on single tissue specimens for healthy subjects, and from patients with lupus nephritis (LN) or diabetic nephropathy (DN). Automated analysis of the aligned tissue images showed enrichment of specific cellular clusters into distinct neighborhoods.

Conclusion

This newly developed imaging method, SeqStain, provides an easy to use and robust platform for deep profiling of kidney tissue specimen. The generated spatial maps will provide important new insights about the disease pathobiology and improve future diagnostics and therapeutics for LN and DN.

Funding

  • NIDDK Support