Copper chelation by tetrathiomolybdate alleviates kidney damage from calcium oxalate stones by inhibiting cuproptosis and oxidative stress

Kidney stone disease (nephrolithiasis), characterized by calcium oxalate (CaOx) deposition, often leads to chronic kidney disease. Recent studies suggest that copper metabolism and oxidative stress play roles in kidney stone pathogenesis. Cuproptosis, a regulated cell death pathway driven by copper, is hypothesized to be implicated in nephrolithiasis. This study aimed to assess the therapeutic potential of tetrathiomolybdate (TTM), a copper chelator, in reducing copper-induced toxicity and oxidative stress in nephrocalcinosis models. Forty C57BL/6 mice were divided into four groups: control, TTM, glyoxylate-induced CaOx, and glyoxylate with TTM. TTM was administered intraperitoneally for 10 days, and kidney function, copper levels, and crystal deposition were assessed. In vitro , human renal epithelial cells (HK-2) treated with CaOx monohydrate were analyzed for reactive oxygen species levels, protein expression, and gene regulation under the TTM treatment. The TTM treatment significantly improved kidney function, reduced copper content, and decreased CaOx crystal deposition in vivo . In vitro , TTM reduced reactive oxygen species, LDH, and malondialdehyde levels in CaOx monohydrate-treated HK-2 cells, while decreasing Cu + level and reversing altered protein expression associated with cuproptosis. A transcriptomic analysis identified four TTM-regulated copper metabolism-related genes: cyclin D1 ( Ccnd1 ), ceruloplasmin ( Cp ), lysyl oxidase ( Lox ), and lox like 2 ( Loxl2 ). TTM mitigates copper-induced toxicity and oxidative stress by modulating cuproptosis-related genes, offering a promising therapeutic approach for nephrolithiasis.