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Kidney Week

Abstract: SA-PO330

Therapeutic Potential of Induced Nephron Progenitor Cells and Their Extracellular Vesicles in Aristolochic Acid-Induced CKD: Insights from Organoid and Mouse Models

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 600 Development, Stem Cells, and Regenerative Medicine

Authors

  • Farry, Justin M., Vanderbilt University Department of Biomedical Engineering, Nashville, Tennessee, United States
  • Bejoy, Julie, Vanderbilt University Division of Nephrology and Hypertension, Nashville, Tennessee, United States
  • Woodard, Lauren Elizabeth, Vanderbilt University Division of Nephrology and Hypertension, Nashville, Tennessee, United States
Background

CKD affects 15% of US adults, with Aristolochic acid (AA) being a valuable model for CKD. We explored CKD therapies using induced nephron progenitor cells (iNPs) and their extracellular vesicles (iNP-EVs). iNPs are engineered HK-2 reprogrammed to a nephron progenitor phenotype using an inducible piggyBac transposon expressing SNAI2, EYA1, and SIX1. We compared the efficacy of iNP cells to iNP-EVs. EVs have similar benefits to injected stem cells while avoiding many associated pitfalls. This study investigated the potential therapeutic effects of iNPs and iNP-EVs on AA-induced renal injury in both iPSC-derived human kidney organoids and animal models.

Methods

Organoids were grown following established protocols and were treated with 10 µM AA for 48 hrs. AA was followed by 24 hrs of exposure to either iNPs or iNP-EVs. In vivo experiments were performed in NSG mice, an immunocompromised mouse strain, to avoid an immune response to iNPs. In brief, mice were injected with 2 mg/kg AA three times per week for two weeks; then, the injury was allowed to develop for an additional two weeks, then mice were treated with iNPs or iNP-EVs.

Results

iNP-treated organoids showed decreased Dickkopf-1 (DKK1) levels compared to EV-treated, untreated, and control organoids. Both iNPs and EVs treatment reduced macrophage colony-stimulating factor (M-CSF) compared to untreated organoids, with EV treatment showing a more pronounced effect. Immunofluorescent staining revealed increased alpha-smooth muscle actin (α-SMA) levels in AA-treated organoids, with lower expression in iNP and EV-treated organoids.

Conclusion

Analysis of tissue sections stained for α-SMA showed that the AA + iNP-EVs treated group had fewer myofibroblasts than the AA-only and AA + iNP-treated groups. All groups receiving AA had more α-SMA staining than the untreated control group. DKK1 is a Wnt signaling inhibitor, suggesting that iNPs may encourage proliferation through a non-exosomal mechanism. The M-CSF reduction suggests an iNP EV-dependent immunomodulatory effect. M-CSF is released from activated fibroblasts, so decreased M-CSF levels could indicate reduced numbers of myofibroblasts, supported by the improvement in α-SMA levels. In conclusion, iNPs and iNP-EVs may mitigate fibrosis in mouse and human models of AA-induced CKD.

Funding

  • Other NIH Support