Abstract: PO1440
SeqStain Is a Novel, Multiplex Imaging Method for Spatialomic Profiling of Human Kidney Tissues
Session Information
- Glomerular Diseases: Immunology and Inflammation in Vasculitis and Lupus Nephritis
November 04, 2021 | Location: On-Demand, Virtual Only
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1202 Glomerular Diseases: Immunology and Inflammation
Authors
- Venkatesh, Ishwarya, Rush University Medical Center, Chicago, Illinois, United States
- Rajagopalan, Anugraha R., Rush University Medical Center, Chicago, Illinois, United States
- Varma, Disha, Rush University Medical Center, Chicago, Illinois, United States
- Gupta, Vineet, Rush University Medical Center, Chicago, Illinois, United States
Group or Team Name
- Team SeqStain
Background
Chronic Kidney Disease (CKD) is the ninth leading cause of death in the United States, an emerging global health challenge affecting 10-15% of the population. Lack of reliable biomarkers precludes early diagnosis of CKD. The progression of CKD is known to be dictated by renal tissue changes. For instance in Diabetic Nephropathy, disease progression is accompanied by considerable changes to the glomeruli, reduced podocyte number, inflammation in the renal tissue, influx of immune cells that ultimately lead to tissue damage. Understanding these tissue-centered events on a deeper level is imperative for early diagnosis
Methods
To understand molecular and cellular composition of tissues and their relative organization in three-dimensional space, we developed a novel multiplex staining method called SeqStain. The SeqStain multiplex platform uses fluorescently labeled DNA oligonucleotides (termed “SeqStain antibodies”) to stain while endonucleases are used to achieve gentle de-staining after each round. This methodology can fluorescently label primary antibodies, secondary antibodies and Fabs of secondary antibodies to efficiently analyze complex tissues. We generated a SeqStain multiplex panel with antibodies that would probe different histological regions relevant to the kidney
Results
Normal kidney was stained with SeqStain antibodies and de-stained using endonucleases. Using SeqStain methodology, we built >20-plex panel on kidney tissue and provided a simple, gentle and rapid technique for multiplex imaging. Strikingly, de-staining using the SeqStain method was rapid and removed ~99% of the signal in <1min without affecting tissue integrity. The method was implemented using a simple perfusion setup with readily available components, allowing staining of tens of antigens on a single tissue section. Alignment of images and their analyses provided spatialomic data on multiple cell types in tissue
Conclusion
The SeqStain method offers a gentle, easy-to-use, and effective multiplex imaging technique that provides a unique platform for obtaining spatialomic insights. SeqStain method can profile the CKD kidney tissues and comprehend the tissue-centered events that could play a role in disease progression. Currently, we are profiling the CKD tissues in multiplex staining experiments in comparison to healthy human kidney to generate spatial maps