Abstract: FR-PO149
Detection of Myeloid Cell Pleiotropy In Situ and the Unique Niches of Immune Cells in Ischemic Kidney Injury Using CODEX Imaging
Session Information
- AKI: Mechanisms - II
November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Sabo, Angela R., Indiana University School of Medicine, Indianapolis, Indiana, United States
- Maghak, Lauren A., Indiana University School of Medicine, Indianapolis, Indiana, United States
- Gulbronson, Connor J., Indiana University School of Medicine, Indianapolis, Indiana, United States
- Khan, Shehnaz, Indiana University School of Medicine, Indianapolis, Indiana, United States
- Micanovic, Radmila, Indiana University School of Medicine, Indianapolis, Indiana, United States
- LaFavers, Kaice Arminda, Indiana University School of Medicine, Indianapolis, Indiana, United States
- Barwinska, Daria, Indiana University School of Medicine, Indianapolis, Indiana, United States
- Williams, James C., Indiana University School of Medicine, Indianapolis, Indiana, United States
- El-Achkar, Tarek M., Indiana University School of Medicine, Indianapolis, Indiana, United States
- Winfree, Seth, University of Nebraska Medical Center, Omaha, Nebraska, United States
Background
Myeloid cells are involved in the pathogenesis of acute kidney injury (AKI), particularly at the early stage. Further, the renal protective protein Uromodulin (UMOD) plays a central role in modulating myeloid populations in the kidney. To understand the myeloid response during AKI in situ and how it is shaped by UMOD, we used CO-Detection by indEXing (CODEX) a cyclic imaging technique that allows for the imaging of more than 30 protein markers in one tissue. Such approach will preserve the spatial context and define the cellular niches that determine the pathogenesis of AKI.
Methods
Whole sections of murine renal tissue from wild-type and UMOD-knockout mice were harvested 6 hours after sham or 22-min ischemic injury and stained for nuclei and 33 different protein markers, targeting parenchymal, immune, and injury proteins. The collected images were processed and stitched using CODEX software and visualized in ImageJ/FIJI. Using nuclei, cells were segmented, associated markers quantitated, and cell types labeled using a semi-supervised tissue cytometry approach in the Volumetric Tissue Exploration and Analysis (VTEA) software.
Results
In sham and injured tissues, our marker panel labeled most cells in the kidney. VTEA cytometry and semi-supervised clustering allowed the identification and quantitation of all major epithelial, endothelial, stromal and immune cells and mapping of these various populations in situ. During AKI, phenotypic pleiotropy of infiltrating and resident myeloid cells was observed and spatially delineated. Using cell-centric neighborhood analysis, unique spatial niches of immune cells associated with specific epithelial and vascular cells were defined. The phenotypic pleiotropy and cells niches were altered by UMOD deficiency.
Conclusion
We establish a robust panel of protein markers to comprehensively capture the cellular diversity in murine kidneys and establish an analyticalan analytical pipeline to uncover unique niches of immune/epithelial/vascular cells in injury. This approach will allow us to uncover the cell-cell interactions that underlie the pathogenesis of AKI and explain the protective role of Uromodulin during injury.
Funding
- NIDDK Support