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Abstract: PO2437

Single-Nucleus RNA-Sequencing Analysis Elucidates Intercellular Interactions Between Inflammatory Parenchymal Cells and Immune Cells in the Kidneys with Tertiary Lymphoid Tissues

Session Information

Category: CKD (Non-Dialysis)

  • 2103 CKD (Non-Dialysis): Mechanisms

Authors

  • Yoshikawa, Takahisa, 1) Department of Nephrology, Graduate School of Medicine, Kyoto University, Kyoto, Japan
  • Oguchi, Akiko, 1) Department of Nephrology, Graduate School of Medicine, Kyoto University, Kyoto, Japan
  • Sato, Yuki, 1) Department of Nephrology, Graduate School of Medicine, Kyoto University, Kyoto, Japan
  • Yamamoto, Takuya, 3) Department of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan
  • Murakawa, Yasuhiro, 2) RIKEN Center for Integrative Medical Sciences, Yokohama, Japan
  • Yanagita, Motoko, 1) Department of Nephrology, Graduate School of Medicine, Kyoto University, Kyoto, Japan
Background

After acute kidney injury (AKI), elderly patients sometimes fail to recover and develop chronic kidney disease (CKD). We previously demonstrated that tertiary lymphoid tissues (TLTs) are formed in aged kidneys after AKI, resulting in prolonged inflammation and impaired regeneration, leading to CKD. However, the mechanism of TLT formation in aged kidneys remains unknown.

Methods

Single nucleus RNA-sequencing (snRNA-seq) for three aged murine kidneys 30 days after ischemic reperfusion injury (IRI) as well as an aged kidney after sham operation was performed using the 10X platform. Computational analysis including subset analysis, ligand-receptor analysis, and pseudotime trajectory analysis were performed. Gene expression was validated by immunostaining and high sensitivity in situ hybridization.

Results

SnRNA-seq generated 15968 and 7485 nuclei transcriptomics from IRI kidneys and a sham kidney, respectively, and demonstrated heterogeneous cell populations including parenchymal cells and immune cells in IRI kidneys with TLTs. We identified a subset of proinflammatory proximal tubules with sustained injury in IRI kidneys with TLTs, and confirmed that some of them surrounded TLTs. Ligand-receptor analysis suggested that this proinflammatory proximal tubules intensively interact with immune cells and fibroblasts. We also identified subsets of profibrotic fibroblasts and proinflammatory fibroblasts, and validated that the latter population being TLT-associated fibroblasts. Pseudotime trajectory analysis of fibroblasts showed that these two types of fibroblasts acquired distinct transcription factors and gene expression patterns along differentiation into distinct populations. Ligand-receptor analysis suggested that proinflammatory fibroblasts possibly contribute to survival and proliferation of B cells through BAFF production in TLT formation.

Conclusion

SnRNA-seq elucidated proinflammatory populations both in proximal tubules and fibroblasts in aged injured kidneys. Various intercellular interactions between the proinflammatory parenchymal cells and immune cells might contribute to TLT formation and these interactions have the potential to be promising therapeutic targets for AKI to CKD transition in the elderly.

Funding

  • Government Support – Non-U.S.