Ascorbate protects human kidney organoids from damage induced by cell-free hemoglobin.

Sepsis-associated acute kidney injury is associated with high morbidity and mortality in critically ill patients. Cell-free hemoglobin (CFH) is released into the circulation of patients with severe sepsis and the levels of CFH are independently associated with mortality. CFH treatment increased cytotoxicity in the human tubular epithelial cell line HK-2. To better model the intact kidney, we cultured human kidney organoids derived from induced pluripotent stem cells. We treated human kidney organoids grown by both three-dimensional (3D) and transwell protocols with CFH for 48 hours. We found evidence for increased tubular toxicity, oxidative stress, mitochondrial fragmentation, endothelial cell injury, and injury-associated transcripts as compared to the untreated control group. To evaluate the protective effect of clinically available small molecules, we co-treated CFH-injured organoids with ascorbate or acetaminophen for 48 hours. We found significantly decreased toxicity, preservation of endothelial cells, and reduced mitochondrial fragmentation in the group receiving Vitamin C (ascorbate) following CFH treatment. This study provides direct evidence that ascorbate or ascorbic acid protects human kidney cells from CFH-induced damage such as occurs in sepsis-associated acute kidney injury.