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Kidney Week

Abstract: PO2468

Single-Nucleus Transcriptional Profiling of CKD After Repeated Low-Dose Cisplatin Treatment

Session Information

Category: CKD (Non-Dialysis)

  • 2103 CKD (Non-Dialysis): Mechanisms

Authors

  • Ma, Zhengwei, Augusta University Medical College of Georgia, Augusta, Georgia, United States
  • Hu, Xiaoru, Augusta University Medical College of Georgia, Augusta, Georgia, United States
  • Dong, Zheng, Augusta University Medical College of Georgia, Augusta, Georgia, United States
Background

Cisplatin induces both acute and chronic kidney problems during chemotherapy. Recent studies have established the models of chronic kidney problems after repeated low dose cisplatin treatment (RLDC). RLDC-induced global transcriptional changes in specific renal cells associated with the development of chronic kidney problems is unclear.

Methods

Male C57BL/6 mice were given 4 consecutive weekly injections of 8 mg/kg cisplatin. Renal function was measured at 5 and 9 weeks after the first cisplatin injection. Kidney tissues were collected for histology and single-nucleus RNA sequencing (Sn-RNA-seq). Cell-type-specific changes in gene expression were compared between the samples from control and cisplatin treated mice. Transcriptional regulators in proximal tubular cells were identified and qPCR was used to validate the critical genes involved in renal fibrotic and inflammation.

Results

RLDC induced decreases in eGFR and kidney weight in mice at 5 and 9 weeks. The kidneys of these mice showed tubular degeneration and dilation. There was also increases in KIM-1 positive tubules and atubular glomeruli. Sn-RNA-seq identified transcripts corresponding to 23021 genes. The markers for 11 cell types and 12 cell clusters were detected. Cluster-by-cluster comparison demonstrated cell-type-specific changes in gene expression that are important for transport, fibrosis and inflammation in RLDC mouse kidneys. In particular, compared with the untreated control, RLDC resulted in 425 differentially expressed genes (log2FC>1, p<0.05) in proximal tubular cells. >400 and >300 genes displaying altered expression were enriched in profibrotic and proinflammatory pathways, respectively. Consistently, RLDC induced NF-κB activation and proinflammatory cytokines (TNFa, IL6 and IL7), and the expression of fibrosis markers (fibronectin, collagen I, vimentin and α-SMA). Furthermore, Runx1 and Spp1 were identified as critical transcriptional factors that drive inflammation and fibrosis progression after repeated cisplatin treatment.

Conclusion

Single-nucleus RNA sequencing revealed altered gene expression and identified critical transcriptional regulators that promote renal inflammation and fibrosis during the development of chronic kidney problems after repeated low dose cisplatin treatment.

Funding

  • NIDDK Support