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Kidney Week

Abstract: PO0334

Cisplatin AKI: Localization of Cell State Injury Clusters with Spatial Transcriptomics

Session Information

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Maddatu, Judith, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Melo ferreira, Ricardo, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Cheng, Yinghua, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Lake, Blue, University of California San Diego, La Jolla, California, United States
  • Menon, Rajasree, University of Michigan Michigan Medicine, Ann Arbor, Michigan, United States
  • Kretzler, Matthias, University of Michigan Michigan Medicine, Ann Arbor, Michigan, United States
  • Jain, Sanjay, Washington University in St Louis School of Medicine, St Louis, Missouri, United States
  • Eadon, Michael T., Indiana University School of Medicine, Indianapolis, Indiana, United States

Group or Team Name

  • the KPMP Consortium
Background

Human acute kidney injury (AKI) is a multifactorial process and severity varies between individuals. The murine cisplatin model is a replicable example of AKI with limited confounding variables. We determined the pathways and injury markers dysregulated across the nephron in cisplatin AKI. We localized relevant human cell state signatures with spatial transcriptomics (ST) to determine which were most prevalent in cisplatin AKI and where these cells reside.

Methods

Six 129S6-SVE mice received vehicle or cisplatin (0.5 mg/g). After 72 hours, mouse kidneys were harvested and preserved for histology, single cell sequencing (scRNAseq), and ST. Gene mapping was completed with Cell Ranger 5.0.1 (scRNAseq) or Space Ranger 1.2.0 (ST). Human scRNAseq data was downloaded from kpmp.org, clustered by cell state (injured, adaptive, degenerative, transitioning, cycling, or reference) and mapped to mouse orthologs in Enembl database. Seurat clustered and integrated scRNAseq data as well as performed spatial mapping. Visualizations were created with R Studio.

Results

Cisplatin mice had increased BUN and tubular atrophy. scRNAseq identified 32 cell clusters, merged across common cell types. Cell cycle genes (Cdkn1a) were upregulated across epithelial cell types, but injury markers (Lcn2 and Spp1) were only upregulated in the distal nephron. A novel cluster of dedifferentiated injured cells was identified and defined by cell cycle markers, Lcn2 and Spp1, suggesting a distal nephron origin despite dedifferentiation. Cdkn1a was upregulated in ST across all cell types, but the novel injury cluster mapped only to the outer renal cortex. From the 6 identified human cell states, the adaptive epithelial state was found to be upregulated in cisplatin mice.

Conclusion

This study localizes the differential effects of cisplatin across the nephron. It adds support to the human injury cell states defined from scRNAseq of heterogenous human samples, showing their relevance in a well-defined AKI model.