Modeling acute kidney injury in kidney organoids with cisplatin

Abstract

Acute kidney injury (AKI) remains a major global healthcare problem and there is a need to develop human-based models to study AKI in vitro. Towards this goal, we have characterized induced pluripotent stem cell-derived human kidney organoids and their response to cisplatin, a chemotherapeutic drug that induces AKI and preferentially damages the proximal tubule. We found that a single treatment with 50 µM cisplatin induces HAVCR1 and CXCL8 expression, DNA damage (γH2AX) and cell death in the organoids in a dose-dependent manner but greatly impairs organoid viability. DNA damage was not specific to the proximal tubule but also affected the distal tubule and interstitial populations. This lack of specificity correlated with low expression of the proximal tubule-specific SLC22A2/OCT2 transporter for cisplatin. To improve viability, we developed a repeated low-dose regimen of 4x 5 µM cisplatin over 7 days and found this causing less toxicity while still inducing a robust AKI response that included secretion of known AKI biomarkers and inflammatory cytokines. This work validates the use of human kidney organoids to model aspects of AKI in vitro, with the potential to identify new AKI biomarkers and develop better therapies.

Tissue Culture Models of AKI: From Tubule Cells to Human Kidney Organoids

“…This is consistent with the acute toxicity, high mortality, and absence of fibrosis found in animal models at higher doses 116(preprint) , 117 . Digby et al also showed that a repeated low-dose regimen of cisplatin avoided acute cell death and resulted in a milder but cumulative injury phenotype compared with a single high dose in kidney organoids 116(preprint) . Such low-dose regimens better mimic the nephrotoxicity found in patients receiving chemotherapy.…”

“…116 In the case of 3D organoids, a higher 50 µM dose of cisplatin induced both HAVCR1/KIM-1 and CXCL8 expression as well as DNA damage and cell death in the organoids. 116 Transcriptional profiling of the corresponding injured organoids showed increased expression of injury biomarkers such as CXCL2, CCL2, HAVCR1, and NGAL with high enrichment in TNF-α signaling. 116 The study suggested that cisplatin-induced kidney organoid damage might be mediated through TNF-α receptors (Fig.…”

“…116 Transcriptional profiling of the corresponding injured organoids showed increased expression of injury biomarkers such as CXCL2, CCL2, HAVCR1, and NGAL with high enrichment in TNF-α signaling. 116 The study suggested that cisplatin-induced kidney organoid damage might be mediated through TNF-α receptors (Fig. 2).…”

“…116,117 Digby et al also showed that a repeated low-dose regimen of cisplatin avoided acute cell death and resulted in a milder but cumulative injury phenotype compared to a single high dose in kidney organoids. 116 Such low-dose regimens better mimic the nephrotoxicity found in patients receiving chemotherapy.…”

“…Digby et al reported that 3D kidney organoids treated with a higher dose of 50 μM cisplatin for 48 hours had upregulation of proximal tubule injury marker HAVCR1 and the inflammatory cytokine C-X-C motif chemokine ligand 8 (CXCL8), and DNA damage and cell death 116(preprint) , 117 . This is consistent with the acute toxicity, high mortality, and absence of fibrosis found in animal models at higher doses 116(preprint) , 117 . Digby et al also showed that a repeated low-dose regimen of cisplatin avoided acute cell death and resulted in a milder but cumulative injury phenotype compared with a single high dose in kidney organoids 116(preprint) .…”