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Abstract: SA-PO0746

Granzyme K-Expressing CD8+ T Cells Drive Complement Activation and Tissue Damage in ANCA-Associated Glomerulonephritis

Session Information

Category: Glomerular Diseases

  • 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology

Authors

  • Panzer, Ulf, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Asada, Nariaki, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Wang, Huiying, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Sultana, Zeba, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Khatri, Robin, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Hellmig, Malte, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Engesser, Jonas, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Ginsberg, Pauline, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Paust, Hans-Joachim, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Kaffke, Anna, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Peters, Anett, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Bonn, Stefan, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Huber, Tobias B., Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Freiwald, Tilo, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Zipfel, Peter F., Leibniz-Institut fur Naturstoff-Forschung und Infektionsbiologie eV Hans-Knoll-Institut, Jena, Thuringia, Germany
  • Wiech, Thorsten, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Mittrücker, Hans-willi, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
  • Krebs, Christian F., Universitatsklinikum Hamburg-Eppendorf, Hamburg, Germany
Background

ANCA–associated vasculitis is a systemic autoimmune disease and the most common cause of rapidly progressive GN (ANCA-GN). Recent experimental and clinical studies highlight the critical role of the complement system in the pathogenesis of ANCA-GN, but the underlying molecular mechanisms driving complement activation are unclear. Two recent Nature publications (PMID 39914456, PMID 38826230) discovered a new mechanism of complement activation that is driven exclusively by granzyme K (GZMK), a serine protease that is mainly expressed by T cells. In this study, we aim to decipher the potential role of this pathway in ANCA-GN.

Methods

Here we applied combined single-cell RNA-sequencing and spatial transcriptomic technologies to blood and kidney biopsy samples of 92 ANCA-GN patients and 16 healthy controls. In total 82 thousand T cells were isolated from kidney biopsies of ANCA-GN patients for single-cell RNA+TCR sequencing, and more than 3 million immune and kidney cells were analyzed by high-resolution spatial transcriptomics. In addition, ex vivo T cell activation and complement activation assays were performed.

Results

Using this immune profiling approach, we identified a distinct CD8+ T cell population, characterized by the expression of GZMK, as the main cellular source of GZMK in ANCA-GN patients. These GZMK+ CD8+ T cells were clonally expanded, highly activated and enriched in the kidney. In an independent spatial transcriptomic validation cohort (48 ANCA-GN patients), we confirmed that GZMK+ CD8+ T cells were increased in patients with ANCA-GN and showed that GZMK producing CD8+ T cells are localized in renal areas of complement activation and tissue injury. Using in vitro assays, we finally showed that kidney CD8+ from ANCA-GN patients release GZMK protein upon T cell receptor activation, and that GZMK directly induces complement activation and deposition on the surface of target cells.

Conclusion

These findings decode a previously unidentified mechanism of complement activation in the kidney of patients with ANCA-GN, and identify the CD8+ GZMK+ T cell-mediated pathway as an attractive therapeutic target for the treatment of these patients.

Funding

  • Government Support – Non-U.S.

Digital Object Identifier (DOI)