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Abstract: TH-PO639

Kidney Organoids for Improved Prediction of Human Nephrotoxicity

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 501 Development, Stem Cells, and Regenerative Medicine: Basic

Authors

  • Tamura, Akitoshi, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
  • Susa, Koichiro, Brigham and Women's Hospital, Harvard Medical School, Boston, United States
  • Galichon, Pierre, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
  • Gupta, Navin R., Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
  • Miyoshi, Tomoya, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
  • Sabbisetti, Venkata, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
  • Bonventre, Joseph V., Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
  • Morizane, Ryuji, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
Background

Nephrotoxicity accounts for only 2% of drug development failures in pre-clinical studies which rises to 19% of drug attrition during phase 3 clinical trials, highlighting the need for improved pre-clinical predictive tools. The poor predictability stems either from inter-species differences or from an inability to distinguish drug-specific versus generalized toxicity due to poor expression of drug transporters (TPs), such as Organic Cation Transporters (OCTs) and Organic Anion Transporters (OATs) in primary cells, and limited availability of co-culturing models recapitulating cell-cell interactions in vitro. Here we demonstrate the utility of human pluripotent stem cell-derived kidney organoids for pre-clinical nephrotoxic assessment.

Methods

Kidney organoids were generated following a 6-step directed differentiation protocol. Using qPCR and immunostaining, drug TP expression was characterized overtime from differentiation day 8 - 50. The following known nephrotoxicants were tested: cisplatin (an OCT-mediated tubular toxicant), aristolochic acid and tenofovir (OAT-mediated tubular toxicants), and puromycin aminonucleoside (a Plasma Monoamine Transporters [PMAT]-mediated podocyte toxicant). Injury responses were assessed by immunostaining, qPCR and biomarker assays. Cimetidine and probenecid, inhibitors of OCTs and OATs respectively, were employed to assess drug TP-mediated nephrotoxicity in tubules.

Results

Drug TPs, OAT1, OAT3, OCT2 and PMAT, were expressed in kidney organoids with maturity. Cisplatin 5 μM induced KIM-1 and γH2AX expression in LTL+ tubules which was partially reversed by cimetidine, indicating TP-mediated injury. Cisplatin at 50 μM resulted in widespread injury to all nephron compartments with increased levels of MCP-1 and TNFR1 in culture media, representing generalized toxicity responses. Aristolochic acid and tenofovir also caused LTL+ tubular injury at a low concentration, and probenecid suppressed injury responses. Puromycin aminonucleoside induced loss of foot processes and podocyte apoptosis without evidence of tubular injury.

Conclusion

Kidney organoids replicated drug TP-mediated nephrotoxicity in a segment-specific manner, serving as a novel pre-clinical tool to assess nephrotoxicity with the ability to distinguish between drug-specific and generalized toxicity responses.

Funding

  • NIDDK Support