Glutathione levels are associated with methotrexate resistance in acute lymphoblastic leukemia cell lines

Frontiers in Oncology

Oncology

Ana Carolina de Mattos Zeri¹  Juliana Ronchi Corrêa²  Carolina Pereira de Souza Melo²  Leonardo Luiz Artico²  Silvia Regina Brandalise²  Rafael Renatino Canevarolo²  Nathalia Moreno Cury²  Yanca Tonhasca Lau²  Samara Sousa Mariano²  José Andrés Yunes³  Praneeth Reddy Sudalagunta⁴ 

[1] Brazilian Biosciences National Laboratory (LNBio), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, SP, Brazil;Centro de Pesquisa Boldrini, Centro Infantil Boldrini, Campinas, SP, Brazil;Centro de Pesquisa Boldrini, Centro Infantil Boldrini, Campinas, SP, Brazil;Medical Genetics Department, Faculty of Medical Sciences, State University of Campinas, Campinas, SP, Brazil;Department of Cancer Physiology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, United States;

关键词: acute lymphoblastic leukemia;    methotrexate;    glutathione;    metabolomics;    drug resistance;    arsenic trioxide;    thioredoxin reductase;   

DOI  :  10.3389/fonc.2022.1032336

 received in 2022-08-30, accepted in 2022-11-02,  发布年份 2022

来源: Frontiers

【 摘 要 】

IntroductionMethotrexate (MTX), a folic acid antagonist and nucleotide synthesis inhibitor, is a cornerstone drug used against acute lymphoblastic leukemia (ALL), but its mechanism of action and resistance continues to be unraveled even after decades of clinical use.MethodsTo better understand the mechanisms of this drug, we accessed the intracellular metabolic content of 13 ALL cell lines treated with MTX by 1H-NMR, and correlated metabolome data with cell proliferation and gene expression. Further, we validated these findings by inhibiting the cellular antioxidant system of the cells in vitro and in vivo in the presence of MTX.ResultsMTX altered the concentration of 31 out of 70 metabolites analyzed, suggesting inhibition of the glycine cleavage system, the pentose phosphate pathway, purine and pyrimidine synthesis, phospholipid metabolism, and bile acid uptake. We found that glutathione (GSH) levels were associated with MTX resistance in both treated and untreated cells, suggesting a new constitutive metabolic-based mechanism of resistance to the drug. Gene expression analyses showed that eight genes involved in GSH metabolism were correlated to GSH concentrations, 2 of which (gamma-glutamyltransferase 1 [GGT1] and thioredoxin reductase 3 [TXNRD3]) were also correlated to MTX resistance. Gene set enrichment analysis (GSEA) confirmed the association between GSH metabolism and MTX resistance. Pharmacological inhibition or stimulation of the main antioxidant systems of the cell, GSH and thioredoxin, confirmed their importance in MTX resistance. Arsenic trioxide (ATO), a thioredoxin inhibitor used against acute promyelocytic leukemia, potentiated MTX cytotoxicity in vitro in some of the ALL cell lines tested. Likewise, the ATO+MTX combination decreased tumor burden and extended the survival of NOD scid gamma (NSG) mice transplanted with patient-derived ALL xenograft, but only in one of four ALLs tested.ConclusionAltogether, our results show that the cellular antioxidant defense systems contribute to leukemia resistance to MTX, and targeting these pathways, especially the thioredoxin antioxidant system, may be a promising strategy for resensitizing ALL to MTX.

【 授权许可】

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Copyright © 2022 Canevarolo, Melo, Cury, Artico, Corrêa, Lau, Mariano, Sudalagunta, Brandalise, Zeri and Yunes