Abstract: TH-PO655
Immune-epithelial Niches Are Dose-Dependent in Calcineurin Inhibitor Nephrotoxicity
Session Information
- Transplantation: Basic
November 03, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Transplantation
- 2001 Transplantation: Basic
Authors
- Winfree, Seth, University of Nebraska Medical Center, Omaha, Nebraska, United States
- Mcandrews, Kyle, University of Nebraska Medical Center, Omaha, Nebraska, United States
- Vance, Krysten, University of Nebraska Medical Center, Omaha, Nebraska, United States
- Kubik, Gregory S., University of Nebraska Medical Center, Omaha, Nebraska, United States
- Zmijewska, Anna Alicja, University of Alabama, Birmingham, Alabama, United States
- Eudy, James, University of Nebraska Medical Center, Omaha, Nebraska, United States
- Mannon, Roslyn B., University of Nebraska Medical Center, Omaha, Nebraska, United States
Background
While mainstay immunosuppressive agents in kidney transplantation, calcineurin inhibitors (CNIs) are limited by their intrinsic nephrotoxicity. Understanding the mechanisms of injury following chronic CNI exposure is critical to developing strategies to ameliorate this toxicity. There is growing evidence the interplay between the renal tubular epithelium and immune cells plays a critical role in renal injury and disease. We used spatial transcriptomics (ST) to elucidate this interaction and the role immune-epithelial niches may play in CNI nephrotoxicity.
Methods
8-10 week old mice on a low-salt diet were treated with 90 or 120 mg/Kg Cyclosporine A (CsA) daily for two weeks and kidneys were harvested and frozen. Frozen kidneys were embedded in optimal cutting temperature media prior to sectioning. High quality tissue sections (RIN 6-8) were processed for use on the 10x Visium platform including imaging and library generation followed by next-generation sequencing. FASTQ and TIFF file processing were performed with CellRanger. Expression matrices and images were imported into Seurat (v4.0) for analysis. Cell-type distributions within Visium spots were determined by deconvolution with SPOTLight (v0.1.7) using a scRNASeq reference library generated from mouse kidneys.
Results
Cortical and medullary markers of the murine kidney were identified by ST and spatially correlated with histologically defined regions of the kidney. Following treatment with CsA, kidney injury markers, Havcr1 and Lcn2, increased. At the higher dose of CsA, there was a differential upregulation of immune markers, cytokine/chemokines and profibrotic genes. After Visium spot deconvolution, epithelial cell signatures were identified and localized to expected regions of the kidney. Further, signatures of immune cells were associated with specific tubular subsegments (cortical vs. medullary-thick ascending limb) in a CsA dose dependent manner.
Conclusion
Our data demonstrates ST can uncover molecular profiles and cellular niches of CNI nephrotoxicity. Furthermore, our data provide initial observations of a shift in the transcriptional state of the kidney and the cellular niches of toxicity with increasing doses of CsA. This suggests multiple mechanisms of injury in nephrotoxicity may be at play as a function of CNI dose.
Funding
- Veterans Affairs Support