The Expression and Activation of the NF-κB Pathway Correlate with Methotrexate Resistance and Cell Proliferation in Acute Lymphoblastic Leukemia

Overview

Journal Genes (Basel)

Publisher MDPI

Date 2023 Oct 28

PMID 37895229

Authors

Rafael Renatino Canevarolo

Nathalia Moreno Cury

Jose Andres Yunes

Affiliations

Soon will be listed here.

Abstract

Acute lymphoblastic leukemia (ALL) is the most common childhood cancer. Although its prognosis continually improves with time, a significant proportion of patients still relapse from the disease because of the leukemia's resistance to therapy. Methotrexate (MTX), a folic-acid antagonist, is a chemotherapy agent commonly used against ALL and as an immune-system suppressant for rheumatoid arthritis that presents multiple and complex mechanisms of action and resistance. Previous studies have shown that MTX modulates the nuclear factor kappa B (NF-κB) pathway, an important family of transcription factors involved in inflammation, immunity, cell survival, and proliferation which are frequently hyperactivated in ALL. Using a gene set enrichment analysis of publicly available gene expression data from 161 newly diagnosed pediatric ALL patients, we found the (TNF-α) via to be the most enriched Cancer Hallmark in MTX-poor-responder patients. A transcriptomic analysis using a panel of ALL cell lines (six B-cell precursor acute lymphoblastic leukemia and seven T-cell acute lymphoblastic leukemia) also identified the same pathway as differentially enriched among MTX-resistant cell lines, as well as in slowly dividing cells. To better understand the crosstalk between NF-κB activity and MTX resistance, we genetically modified the cell lines to express luciferase under an NF-κB-binding-site promoter. We observed that the fold change in NF-κB activity triggered by TNF-α (but not MTX) treatment correlated with MTX resistance and proliferation across the lines. At the individual gene level, expression was directly associated with a poorer clinical response to MTX and with both an increased TNF-α-triggered NF-κB activation and MTX resistance in the cell lines. Despite these results, the pharmacological inhibition (using BAY 11-7082 and parthenolide) or stimulation (using exogenous TNF-α supplementation) of the NF-κB pathway did not alter the MTX resistance of the cell lines significantly, evidencing a complex interplay between MTX and NF-κB in ALL.

References

Spurlock 3rd C, Gass 4th H, Bryant C, Wells B, Olsen N, Aune T . Methotrexate-mediated inhibition of nuclear factor κB activation by distinct pathways in T cells and fibroblast-like synoviocytes. Rheumatology (Oxford). 2014; 54(1):178-87. PMC: 4269792. DOI: 10.1093/rheumatology/keu279. View

Bosman M, Schuringa J, Vellenga E . Constitutive NF-κB activation in AML: Causes and treatment strategies. Crit Rev Oncol Hematol. 2015; 98:35-44. DOI: 10.1016/j.critrevonc.2015.10.001. View

Spurlock 3rd C, Tossberg J, Matlock B, Olsen N, Aune T . Methotrexate inhibits NF-κB activity via long intergenic (noncoding) RNA-p21 induction. Arthritis Rheumatol. 2014; 66(11):2947-57. PMC: 4211976. DOI: 10.1002/art.38805. View

Kordes U, Krappmann D, Heissmeyer V, Ludwig W, Scheidereit C . Transcription factor NF-kappaB is constitutively activated in acute lymphoblastic leukemia cells. Leukemia. 2000; 14(3):399-402. DOI: 10.1038/sj.leu.2401705. View

Gilmore T . The Rel/NF-kappaB signal transduction pathway: introduction. Oncogene. 1999; 18(49):6842-4. DOI: 10.1038/sj.onc.1203237. View

Zhou X, Li Z, Zhou J . Tumor necrosis factor α in the onset and progression of leukemia. Exp Hematol. 2016; 45:17-26. DOI: 10.1016/j.exphem.2016.10.005. View

Ghashghaeinia M, Toulany M, Saki M, Bobbala D, Fehrenbacher B, Rupec R . The NFĸB pathway inhibitors Bay 11-7082 and parthenolide induce programmed cell death in anucleated Erythrocytes. Cell Physiol Biochem. 2011; 27(1):45-54. DOI: 10.1159/000325204. View

Hewamana S, Lin T, Rowntree C, Karunanithi K, Pratt G, Hills R . Rel a is an independent biomarker of clinical outcome in chronic lymphocytic leukemia. J Clin Oncol. 2009; 27(5):763-9. DOI: 10.1200/JCO.2008.19.1114. View

Tsaouli G, Barbarulo A, Vacca A, Screpanti I, Felli M . Molecular Mechanisms of Notch Signaling in Lymphoid Cell Lineages Development: NF-κB and Beyond. Adv Exp Med Biol. 2020; 1227:145-164. DOI: 10.1007/978-3-030-36422-9_10. View

10.

Subramanian A, Tamayo P, Mootha V, Mukherjee S, Ebert B, Gillette M . Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A. 2005; 102(43):15545-50. PMC: 1239896. DOI: 10.1073/pnas.0506580102. View

11.

Oeckinghaus A, Ghosh S . The NF-kappaB family of transcription factors and its regulation. Cold Spring Harb Perspect Biol. 2010; 1(4):a000034. PMC: 2773619. DOI: 10.1101/cshperspect.a000034. View

12.

Qin H, Wilson C, Lee S, Zhao X, Benveniste E . LPS induces CD40 gene expression through the activation of NF-kappaB and STAT-1alpha in macrophages and microglia. Blood. 2005; 106(9):3114-22. PMC: 1895321. DOI: 10.1182/blood-2005-02-0759. View

13.

Kalmanti M, Karamolengou K, Dimitriou H, Tosca A, Vlachonikolis I, Peraki M . Serum levels of tumor necrosis factor and soluble interleukin 2 receptor as markers of disease activity and prognosis in childhood leukemia and lymphoma. Int J Hematol. 1993; 57(2):147-52. View

14.

Cleaver A, Beesley A, Firth M, Sturges N, OLeary R, Hunger S . Gene-based outcome prediction in multiple cohorts of pediatric T-cell acute lymphoblastic leukemia: a Children's Oncology Group study. Mol Cancer. 2010; 9:105. PMC: 2879253. DOI: 10.1186/1476-4598-9-105. View

15.

Kamienska E, Ociepa T, Wysocki M, Kurylak A, Matysiak M, Urasinski T . Activation of NF-ĸB in leukemic cells in response to initial prednisone therapy in children with acute lymphoblastic leukaemia: relation to other prognostic factors. Pol J Pathol. 2011; 62(1):5-11. View

16.

Chandel N, Trzyna W, McClintock D, Schumacker P . Role of oxidants in NF-kappa B activation and TNF-alpha gene transcription induced by hypoxia and endotoxin. J Immunol. 2000; 165(2):1013-21. DOI: 10.4049/jimmunol.165.2.1013. View

17.

Guldenpfennig C, Teixeiro E, Daniels M . NF-kB's contribution to B cell fate decisions. Front Immunol. 2023; 14:1214095. PMC: 10391175. DOI: 10.3389/fimmu.2023.1214095. View

18.

Mercogliano M, Bruni S, Elizalde P, Schillaci R . Tumor Necrosis Factor α Blockade: An Opportunity to Tackle Breast Cancer. Front Oncol. 2020; 10:584. PMC: 7189060. DOI: 10.3389/fonc.2020.00584. View

19.

Vilimas T, Mascarenhas J, Palomero T, Mandal M, Buonamici S, Meng F . Targeting the NF-kappaB signaling pathway in Notch1-induced T-cell leukemia. Nat Med. 2006; 13(1):70-7. DOI: 10.1038/nm1524. View

20.

Dos Santos N, Ghezzo M, da Silva R, Fernandes M . NF-κB in T-cell Acute Lymphoblastic Leukemia: Oncogenic Functions in Leukemic and in Microenvironmental Cells. Cancers (Basel). 2013; 2(4):1838-60. PMC: 3840450. DOI: 10.3390/cancers2041838. View