ASN's Mission

ASN leads the fight to prevent, treat, and cure kidney diseases throughout the world by educating health professionals and scientists, advancing research and innovation, communicating new knowledge, and advocating for the highest quality care for patients.

learn more

Contact ASN

1401 H St, NW, Ste 900, Washington, DC 20005

email@asn-online.org

202-640-4660

The Latest on Twitter

Kidney Week

Abstract: PO0391

Modeling Kidney Injury and Repair in Kidney Organoids Reveals an Intrinsic Repair Mechanism

Session Information

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Gupta, Navin R., Massachusetts General Hospital, Boston, Massachusetts, United States
  • Matsumoto, Takuya, Harvard Medical School, Boston, Massachusetts, United States
  • Hiratsuka, Ken, Massachusetts General Hospital, Boston, Massachusetts, United States
  • Galichon, Pierre, Brigham and Women's Hospital, Boston, Massachusetts, United States
  • Miyoshi, Tomoya, Brigham and Women's Hospital, Boston, Massachusetts, United States
  • Susa, Koichiro, Brigham and Women's Hospital, Boston, Massachusetts, United States
  • Tatsumoto, Narihito, Cedars-Sinai Medical Center, Los Angeles, California, United States
  • Yamashita, Michifumi, Cedars-Sinai Medical Center, Los Angeles, California, United States
  • Morizane, Ryuji, Massachusetts General Hospital, Boston, Massachusetts, United States
Background

Kidneys have the capacity for intrinsic repair, preserving kidney architecture with return to a basal state following tubular injury. When injury is overwhelming or repetitive, that capacity is exceeded and incomplete repair results in scar tissue replacing normal kidney parenchyma. Loss of nephrons correlates with reduced kidney function, which defines chronic kidney disease (CKD) and confers significant morbidity and mortality to the worldwide population. Despite the identification of pathways involved in intrinsic repair, limited treatments for CKD exist, in part due to the limited throughput and predictivity of animal studies.

Methods

hPSC-derived kidney organoids were subject to repeated cisplatin injury twice weekly from differentiation days 49 to 63. Samples were harvested following each injury for immunostaining and qPCR to determine transition from intrinsic to incomplete repair. Single nuclear sequencing (snRNAseq) of pooled samples of control, intrinsic repair, and incomplete repair were compared to similar data sets of mouse UUO and IRI, and human rejecting kidney transplant. Transcriptomic results were validated with fibrotic human kidney biopsy samples by immunostaining. Targeted drug screening was conducted in kidney organoids to promote intrinsic repair for the identification of novel therapeutic candidates.

Results

snRNA-seq from kidney organoids identified 159 differentially expressed genes and 29 altered signal pathways during intrinsic repair. Tubular atrophy and the induction of scar-forming myofibroblasts correlates with reduced expression of homology-directed repair genes in injured tubular cells, a finding supported by single cellular transcriptomics in models of obstructive, hemodynamic, and immune-mediated kidney injury, as well as biopsy samples of patients with fibrotic kidney disease. We identified FANCD2/RAD51-mediated repair as a critical determinant governing the transition between intrinsic and incomplete repair and identified a novel therapeutic target for the prevention of CKD onset and progression following AKI.

Conclusion

Our findings demonstrate the utility of kidney organoids in determining novel pathologic pathways, conducting mechanistic studies of human kidney disease and identifying druggable targets through translational studies.

Funding

  • NIDDK Support