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Kidney Week

Abstract: TH-PO420

Genome Therapy in Kidney Organoids Reveals that Canonical NFκB Signaling Governs Tubular Senescence, Cytotoxicity, and Immunogenicity in Response to Adeno-Associated Viruses (AAV)

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 600 Development, Stem Cells, and Regenerative Medicine

Authors

  • Gupta, Navin R., Mayo Clinic Minnesota, Rochester, Minnesota, United States
  • Oishi, Haruka, Massachusetts General Hospital, Boston, Massachusetts, United States
  • Inomata, Kenta, Massachusetts General Hospital, Boston, Massachusetts, United States
  • Morizane, Ryuji, Massachusetts General Hospital, Boston, Massachusetts, United States
Background

Genome therapy is revolutionizing the treatment of inherited diseases by directly editing the human genome. Due to its potential to irreversibly alter human DNA, systemic gene therapy raises significant concerns, demanding a new approach to preclinical evaluations. These concerns are further illustrated by recent tragedies in clinical trials, which have led to organ failures and patient fatalities. Due to high blood flow, the kidney emerges as a focal point for drug-related toxicity.

Methods

Human stem cell-derived kidney organoids are nephron-rich, highly structured 3-dimensional tissue capable of exocrine and endocrine functions, have modeled human genetic and non-genetic disease, kidney injury and repair, and nephrotoxicity testing. Here we use kidney organoids as a pre-clinical platform for CRISPR genome editing via adeno-associated virus (AAV) delivery, a common genome editing strategy in clinical trials that have raised safety concerns. Although AAV is of limited pathogenesis in humans, clinical use has required supraphysiologic titers often met with toxicity.

Results

Kidney organoids validated the tropism of differing AAV serotypes for human kidney. Applying immunostaining to organoid cryosections and electrochemiluminescence-based mesoscale discovery to their supernatants, we found that AAV drives tubular genotoxicity, cytotoxicity, and immunogenicity. RNA-sequencing identified an association between tubular injury and IL-1β activation of the canonical NFκB pathway. Targeted inhibition of canonical NFκB signaling prevented the fibrotic loss of nephrons by ameliorating p21+ senescence in injured tubules.

Conclusion

Our findings demonstrate the translational utility of kidney organoids as a pre-clinical testing platform for genome therapy, which may inform clinical trials of strategies that limit organ failures and death.

Funding

  • NIDDK Support