Abstract: FR-PO1093
Single-Nucleus RNA Sequencing Reveals Cellular Heterogeneity and Cell-Cell Interactions Contributing to Tertiary Lymphoid Tissue Development in the Aged Injured Kidney
Session Information
- CKD Mechanisms: Progression, Fibrosis, and Beyond
November 03, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: CKD (Non-Dialysis)
- 2303 CKD (Non-Dialysis): Mechanisms
Authors
- Yoshikawa, Takahisa, Department of Nephrology, Graduate School of Medicine, Kyoto University, Kyoto, Kyoto, Japan
- Oguchi, Akiko, Department of Nephrology, Graduate School of Medicine, Kyoto University, Kyoto, Kyoto, Japan
- Toriu, Naoya, Department of Nephrology, Graduate School of Medicine, Kyoto University, Kyoto, Kyoto, Japan
- Sato, Yuki, Department of Nephrology, Graduate School of Medicine, Kyoto University, Kyoto, Kyoto, Japan
- Yamamoto, Takuya, Department of Life Science Frontiers, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Kyoto, Japan
- Murakawa, Yasuhiro, Institute for the Advanced Study of Human Biology (WPI-ASHBi), Kyoto University, Kyoto, Kyoto, Japan
- Yanagita, Motoko, Department of Nephrology, Graduate School of Medicine, Kyoto University, Kyoto, Kyoto, Japan
Group or Team Name
- Yanagita Group.
Background
Tertiary lymphoid tissues (TLTs) are ectopic lymphoid structures that develop in non-lymphoid organs with chronic inflammation. TLTs can develop in kidney diseases such as IgA nephropathy and transplanted kidneys and are associated with poor renal prognosis. However, how TLTs affect renal tissues and expand remains unclear.
Methods
Single-nucleus RNA-sequencing (snRNA-seq) was performed on three aged mouse kidneys with TLTs after ischemia–reperfusion injury or an aged kidney after sham surgery. The results were validated using immunostaining, in situ hybridization (ISH) of murine and human kidneys, and in vitro experiments.
Results
snRNA-seq identified 15,986 and 7,485 nuclei in the injured kidneys and the sham-treated kidney, respectively. In the injured kidneys, proinflammatory and profibrotic Vcam1+ injured proximal tubules (PTs) with activation of NFκB and interferon (IFN)-inducible transcription factors were detected. Importantly, VCAM1+ PTs were preferentially localized around TLTs and suggested to drive inflammation and fibrosis via the expression of multiple chemokines or cytokines. snRNA-seq analysis and ISH revealed that Tnf and Ifng were highly expressed by lymphocytes within TLTs, and these inflammatory cytokines synergistically upregulated VCAM1 and chemokine expression in cultured PT cells. In addition, snRNA-seq and immunostaining identified proinflammatory and profibrotic fibroblasts, which resided within and outside TLTs, respectively. Proinflammatory fibroblasts with STAT1 activation expressed various chemokines or cytokines, including Cxcl9/Cxcl10 and Tnfsf13b (encoding B cell-activating factor), suggesting their contribution to lymphocyte recruitment and survival. IFNγ upregulated the expression of these molecules in cultured fibroblasts in a STAT1-dependent manner, indicating potential bidirectional interactions between proinflammatory fibroblasts and IFNγ-producing CXCR3+ T cells within TLTs. These cellular and molecular components were confirmed in human transplanted kidneys with TLTs.
Conclusion
TLTs develop via intense interactions between proinflammatory renal parenchymal and immune cells. These interactions may serve as novel therapeutic targets in kidney diseases with TLT formation.
Funding
- Government Support – Non-U.S.