Pharmacometabolomics reveals urinary diacetylspermine as a biomarker of doxorubicin effectiveness in triple negative breast cancer

Abstract

Triple-negative breast cancer (TNBC) patients receive chemotherapy treatment, including doxorubicin, due to the lack of targeted therapies. Drug resistance is a major cause of treatment failure in TNBC and therefore, there is a need to identify biomarkers that determine effective drug response. A pharmacometabolomics study was performed using doxorubicin sensitive and resistant TNBC patient-derived xenograft (PDX) models to detect urinary metabolic biomarkers of treatment effectiveness. Evaluation of metabolite production was assessed by directly studying tumor levels in TNBC-PDX mice and human subjects. Metabolic flux leading to biomarker production was determined using stable isotope-labeled tracers in TNBC-PDX ex vivo tissue slices. Findings were validated in 12-h urine samples from control (n = 200), ER+/PR+ (n = 200), ER+/PR+/HER2+ (n = 36), HER2+ (n = 81) and TNBC (n = 200) subjects. Diacetylspermine was identified as a urine metabolite that robustly changed in response to effective doxorubicin treatment, which persisted after the final dose. Urine diacetylspermine was produced by the tumor and correlated with tumor volume. Ex vivo tumor slices revealed that doxorubicin directly increases diacetylspermine production by increasing tumor spermidine/spermine N1-acetyltransferase 1 expression and activity, which was corroborated by elevated polyamine flux. In breast cancer patients, tumor diacetylspermine was elevated compared to matched non-cancerous tissue and increased in HER2+ and TNBC compared to ER+ subtypes. Urine diacetylspermine was associated with breast cancer tumor volume and poor tumor grade. This study describes a pharmacometabolomics strategy for identifying cancer metabolic biomarkers that indicate drug response. Our findings characterize urine diacetylspermine as a non-invasive biomarker of doxorubicin effectiveness in TNBC.

Elevated N1-Acetylspermidine Levels in Doxorubicin-treated MCF-7 Cancer Cells: Histone Deacetylase 10 Inhibition with an N1-Acetylspermidine Mimetic

“…A recent study highlighted the relevance of elevated levels of acetylated polyamines such as diacetylspermine and N1-acetylspermdime in the case of DOX-treated TNBC patient-derived xenograft models. Besides breast cancer, the significance of acetylated forms of polyamines such as N1-acetylspermidine is linked with non-small-cell lung cancer and colorectal cancer [ 44 - 49 ]. Our observation of the elevated intracellular presence of N1-acetylspermidine in DOX-treated breast cancer cells is in line with the existing views of drug-induced metabolic adaptations and polyamines.…”

“…The role of SAT1 has been explored in the drug-induced metabolic adaptations in breast cancer cells [ 46 - 50 ]. Conversely, tumor suppressor SAT1, known for the acetylation of polyamines such as spermidine, could be implicated in drug-induced cell death in cancer cells [ 44 - 49 ].…”

Review on analytical technologies and applications in metabolomics

“…The identification of metabolic pathways affected by a specific drug treatment through metabolomics, as well as a subsequent analysis of genetic variations within these metabolic pathway components could help to achieve personalized medicine [138,142,143]. Profiling of drug-induced changes in metabolites or creating metabolite signatures of different disease subtypes, such as invasive or non-invasive tumors, through high-resolution targeted metabolomics tools serves an important biomarker discovery and diagnostic tool development [143][144][145].…”

Pharmacometabolomics in Drug Disposition, Toxicity and Precision Medicine

“…A major challenge in treating TNBC is that some tumors are resistant to doxorubicin mediated chemotherapy. Untargeted metabolomics were used to identify biomarkers of doxorubicin efficacy in patient-derived xenograft mouse models of TNBC that were either resistant or sensitive to doxorubicin (Velenosi et al, 2022). Patient derived xenograft models of TNBC revealed elevated urinary levels of diacetylspermine in TNBC tumors that were sensitive to doxorubicin.…”