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Kidney Week

Abstract: PO0389

Single-Cell Sequencing of Immune Cells in AKI and Renal Fibrosis

Session Information

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Talabani, Bnar, Cardiff University School of Medicine, Cardiff, Cardiff, United Kingdom
  • Chao, Chia-Ter, Cardiff University School of Medicine, Cardiff, Cardiff, United Kingdom
  • Fraser, Donald, Cardiff University School of Medicine, Cardiff, Cardiff, United Kingdom
  • Taylor, Philip R., Cardiff University School of Medicine, Cardiff, Cardiff, United Kingdom

Group or Team Name

  • Myeloid Cell Biology Group ; Welsh Kidney Research Unit
Background

Immune responses help determine outcome following acute kidney injury (AKI) and Chronic Kidney Disease (CKD) progression. Single Cell RNA Sequencing (scRNAseq) provides an unparalleled opportunity to uncover heterogeneity and provide new mechanistic understanding in AKI-CKD. We have performed scRNAseq of immune cells at specific timepoints mimicking human disease pathology in models of AKI and renal fibrosis.

Methods

Kidneys were harvested from three mice at each time point (Figure 1) mimicking disease states in AKI/CKD. Using a cell sorting strategy, CD45+ cells were isolated from whole kidneys and libraries were prepared on the 10X Genomics platform. ScRNASeq was performed using the Illumina NextSeq 550 System. We conducted Genome mapping using Cellranger and zUMIs and downstream expression analysis was carried out using R and Seurat.

Results

21,734 CD45+ve Cells were sequenced in total. Analysis of gene expression profiles delineated transcriptomic profiles in distinct sub-clusters of immune cells across disease states. Comparison of these demonstrated dynamic changes in immune cell compositions, recruitment and patterns of gene expression, in line with an immune response, to AKI, recovery and fibrosis. Heterogenous clustes of macrophages were seen in disease states, revealing an inverse pattern of gene expression when comparing AKI-recovery and AKI-fibrosis.

Conclusion

ScRNASeq has enabled unbiased profiling of gene expression in disease states important in AKI-CKD. We have identified a novel mechanistic target which we are currently pursuing in knock-out gene experiments that impact macrophage phenotype, that we hypothesise, impacts recovery. This presents an exciting opportunity to study the mechanism of renal fibrosis following AKI.

Figure 1: A schematic diagram of aristolochic acid nephropathy (AAN). Acute AAN represents AKI and chronic AAN represents CKD. Blue arrows represent time points at which kidneys were harvested and scRNASeq of CD45+ immune cells performed.

Funding

  • Private Foundation Support