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

Autor(es):Mostrar menos -

Canevarolo, Rafael Renatino ; de Souza Melo, Carolina Pereira ; Cury, Nathalia Moreno ; Artico, Leonardo Luiz ; Correa, Juliana Ronchi ; Lau, Yanca Tonhasca ; Mariano, Samara Sousa ; Sudalagunta, Praneeth Reddy ; Brandalise, Silvia Regina ; de Mattos Zeri, Ana Carolina ; Yunes, Jose Andres

• Tipo de documento: Artigo Científico
• Fonte: FRONTIERS IN ONCOLOGY; v. 12, p. 18-pg., 2022-12-01.

Resumo

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. (AU)

• Processo FAPESP: 12/11952-0 - Resistência ao metotrexato está diretamente associada à concentração de glutationa em linhagens de leucemia linfóide aguda
• Beneficiário: Rafael Renatino Canevarolo
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 14/08247-8 - Estudos pré-clínicos de novos inibidores de DNA metiltransferase em leucemias agudas
• Beneficiário: Nathalia Moreno Cury
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 08/10034-1 - Microambiente da medula óssea e PI3K na resistência a drogas da leucemia linfóide aguda pediátrica
• Beneficiário: José Andrés Yunes
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 07/00952-0 - Padronização da ressonância magnética nuclear para determinação precoce da resistência a quimioterapia na leucemia linfóide aguda pediátrica
• Beneficiário: Ana Carolina de Mattos Zeri
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 09/04167-1 - Metabolômica da resistência ao metotrexato na leucemia linfóide aguda
• Beneficiário: Rafael Renatino Canevarolo
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 17/02400-7 - Estabelecimento de um modelo animal de leucemia linfóide aguda B-derivada com mutação oncogênica do IL7R
• Beneficiário: Juliana Ronchi Corrêa
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 17/03239-5 - IGFBP7 e resistência a dexametasona na Leucemia Linfoide Aguda
• Beneficiário: Leonardo Luís Artico
• Modalidade de apoio: Bolsas no Brasil - Mestrado