Metabolomic profiling reveals the mechanisms underlying the nephrotoxicity of methotrexate in children with acute lymphoblastic leukemia

Abstract

BackgroundMethotrexate is widely recommended as a first‐line treatment for the intensive systemic and consolidation phases of childhood acute lymphoblastic leukemia. However, methotrexate‐induced nephrotoxicity is a severe adverse reaction, of which the mechanisms remain unclear.MethodsAn untargeted metabolomics analysis of serum from childhood acute lymphoblastic leukemia patients with delayed methotrexate excretion, with or without acute kidney injury, was performed to identify altered metabolites and metabolic pathways. An independent external validation cohort and in vitro HK‐2 cell assays further verified the candidate metabolites, and explored the mechanisms underlying the nephrotoxicity of methotrexate.ResultsFour metabolites showed significant differences between normal excretion and delayed excretion, seven metabolites reflected the differences between groups with or without acute kidney injury, and six pathways were finally enriched. In particular, oxidized glutathione was confirmed as a candidate metabolite involved in the toxicity of methotrexate. We further explored the role of glutathione deprivation‐induced ferroptosis on methotrexate cytotoxicity, and it was found that methotrexate overload significantly reduced cell viability, triggered reactive oxygen species and intracellular Fe2+ accumulation, and altered the expression of ferroptosis‐related proteins in HK‐2 cells. These methotrexate‐induced changes were alleviated or reversed by the administration of a ferroptosis inhibitor, further suggesting that ferroptosis promoted methotrexate‐induced cytotoxicity in HK‐2 cells.ConclusionsOur findings revealed complex metabolomic profiles and provided novel insights into the mechanism by which ferroptosis contributes to the nephrotoxic effects of methotrexate.

Repurposing the diuretic benzamil as an anti-osteosarcoma agent that acts by suppressing integrin/FAK/STAT3 signalling and compromising mitochondrial function

“…Taken together, these observations suggest that the combination of cisplatin/methotrexate and benzamil could provide several advantages over cisplatin or methotrexate alone, such as enhanced anticancer activity and reduced risks of renal and cardiac toxicities from high or accumulated cisplatin and methotrexate dosages. 40 - 42 …”

“…Taken together, these observations suggest that the combination of cisplatin/methotrexate and benzamil could provide several advantages over cisplatin or methotrexate alone, such as enhanced anticancer activity and reduced risks of renal and cardiac toxicities from high or accumulated cisplatin and methotrexate dosages. [40][41][42] Integrins are signalling molecules that act upstream of FAK and STAT3. 27,43 Moreover, integrins α5, 44 αV, [45][46][47] and β1 48 are known to be involved in the development of osteosarcoma.…”