IKZF1 and BTG1 Silencing Reduces Glucocorticoid Response in B-cell Precursor Acute Leukemia Cell Line

Overview

Journal Hematol Transfus Cell Ther

Specialty Hematology

Date 2024 Aug 2

PMID 39095315

Authors

Amanda de Albuquerque

Bruno A Lopes

Renan Amphilophio Fernandes

Etel Rodrigues Pereira Gimba

Mariana Emerenciano

Affiliations

Soon will be listed here.

Abstract

Introduction: Secondary genetic alterations, which contribute to the dysregulation of cell cycle progression and lymphoid specialization, are frequently observed in B-cell precursor acute lymphoblastic leukemia (B-ALL). As IKZF1 and BTG1 deletions are associated with a worse outcome in B-ALL, this study aimed to address whether they synergistically promote glucocorticoid resistance.

Methods: Small interfering RNA was used to downregulate either IKZF1, or BTG1, or both genes in the 207 B-ALL cell line. Cell viability was investigated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and trypan blue exclusion assays. The expression levels of IKZF1, BTG1 and glucocorticoid-responsive genes (DUSP1, SGK1, FBXW7 and NR3C1) were evaluated by real time quantitative real time polymerase chain reaction (PCR).

Results: Isolated silencing of BTG1, IKZF1, or both genes in combination under dexamethasone treatment increased cell viability by 24%, 40% and 84%, respectively. Although BTG1 silencing did not alter the expression of glucocorticoid-responsive genes, IKZF1 knockdown decreased the transcript levels of DUSP1 (2.6-fold), SGK1 (1.8-fold), FBXW7 (2.2-fold) and NR3C1 (1.7-fold). The expression of glucocorticoid-responsive genes reached even lower levels (reducing 2.4-4 fold) when IKZF1 and BTG1 silencing occurred in combination.

Conclusions: IKZF1 silencing impairs the transcription of glucocorticoid-responsive genes; this effect is enhanced by concomitant loss of BTG1. These results demonstrate the molecular mechanism by which the combination of both genetic deletions might contribute to higher relapse rates in B-ALL.

References

Asnafi V, Buzyn A, Le Noir S, Baleydier F, Simon A, Beldjord K . NOTCH1/FBXW7 mutation identifies a large subgroup with favorable outcome in adult T-cell acute lymphoblastic leukemia (T-ALL): a Group for Research on Adult Acute Lymphoblastic Leukemia (GRAALL) study. Blood. 2008; 113(17):3918-24. DOI: 10.1182/blood-2008-10-184069. View

Rogers J, Gupta R, Reyes J, Gundry M, Medrano G, Guzman A . Modeling IKZF1 lesions in B-ALL reveals distinct chemosensitivity patterns and potential therapeutic vulnerabilities. Blood Adv. 2021; 5(19):3876-3890. PMC: 8679666. DOI: 10.1182/bloodadvances.2020002408. View

Findley H, Gu L, Yeager A, Zhou M . Expression and regulation of Bcl-2, Bcl-xl, and Bax correlate with p53 status and sensitivity to apoptosis in childhood acute lymphoblastic leukemia. Blood. 1997; 89(8):2986-93. View

Timmermans S, Souffriau J, Libert C . A General Introduction to Glucocorticoid Biology. Front Immunol. 2019; 10:1545. PMC: 6621919. DOI: 10.3389/fimmu.2019.01545. View

Stanulla M, Dagdan E, Zaliova M, Moricke A, Palmi C, Cazzaniga G . IKZF1 Defines a New Minimal Residual Disease-Dependent Very-Poor Prognostic Profile in Pediatric B-Cell Precursor Acute Lymphoblastic Leukemia. J Clin Oncol. 2018; 36(12):1240-1249. DOI: 10.1200/JCO.2017.74.3617. View

Steliarova-Foucher E, Colombet M, Ries L, Moreno F, Dolya A, Bray F . International incidence of childhood cancer, 2001-10: a population-based registry study. Lancet Oncol. 2017; 18(6):719-731. PMC: 5461370. DOI: 10.1016/S1470-2045(17)30186-9. View

Yuniati L, Scheijen B, Van der Meer L, van Leeuwen F . Tumor suppressors BTG1 and BTG2: Beyond growth control. J Cell Physiol. 2018; 234(5):5379-5389. PMC: 6587536. DOI: 10.1002/jcp.27407. View

Abraham S, Lawrence T, Kleiman A, Warden P, Medghalchi M, Tuckermann J . Antiinflammatory effects of dexamethasone are partly dependent on induction of dual specificity phosphatase 1. J Exp Med. 2006; 203(8):1883-9. PMC: 2118371. DOI: 10.1084/jem.20060336. View

Mullighan C, Goorha S, Radtke I, Miller C, Coustan-Smith E, Dalton J . Genome-wide analysis of genetic alterations in acute lymphoblastic leukaemia. Nature. 2007; 446(7137):758-64. DOI: 10.1038/nature05690. View

10.

Kourtis N, Strikoudis A, Aifantis I . Emerging roles for the FBXW7 ubiquitin ligase in leukemia and beyond. Curr Opin Cell Biol. 2015; 37:28-34. PMC: 4687017. DOI: 10.1016/j.ceb.2015.09.003. View

11.

Inaba H, Greaves M, Mullighan C . Acute lymphoblastic leukaemia. Lancet. 2013; 381(9881):1943-55. PMC: 3816716. DOI: 10.1016/S0140-6736(12)62187-4. View

12.

Hamadeh L, Enshaei A, Schwab C, Alonso C, Attarbaschi A, Barbany G . Validation of the United Kingdom copy-number alteration classifier in 3239 children with B-cell precursor ALL. Blood Adv. 2019; 3(2):148-157. PMC: 6341196. DOI: 10.1182/bloodadvances.2018025718. View

13.

Mullighan C, Su X, Zhang J, Radtke I, Phillips L, Miller C . Deletion of IKZF1 and prognosis in acute lymphoblastic leukemia. N Engl J Med. 2009; 360(5):470-80. PMC: 2674612. DOI: 10.1056/NEJMoa0808253. View

14.

Ramkissoon A, Chaney K, Milewski D, Williams K, Williams R, Choi K . Targeted Inhibition of the Dual Specificity Phosphatases DUSP1 and DUSP6 Suppress MPNST Growth via JNK. Clin Cancer Res. 2019; 25(13):4117-4127. PMC: 6606396. DOI: 10.1158/1078-0432.CCR-18-3224. View

15.

Webster M, Goya L, Ge Y, Maiyar A, Firestone G . Characterization of sgk, a novel member of the serine/threonine protein kinase gene family which is transcriptionally induced by glucocorticoids and serum. Mol Cell Biol. 1993; 13(4):2031-40. PMC: 359524. DOI: 10.1128/mcb.13.4.2031-2040.1993. View

16.

Volk K, Stokes J . SGK is a primary glucocorticoid-induced gene in the human. J Steroid Biochem Mol Biol. 2001; 75(1):51-6. DOI: 10.1016/s0960-0760(00)00136-9. View

17.

van Galen J, Kuiper R, van Emst L, Levers M, Tijchon E, Scheijen B . BTG1 regulates glucocorticoid receptor autoinduction in acute lymphoblastic leukemia. Blood. 2010; 115(23):4810-9. DOI: 10.1182/blood-2009-05-223081. View

18.

Schmidt S, Rainer J, Riml S, Ploner C, Jesacher S, Achmuller C . Identification of glucocorticoid-response genes in children with acute lymphoblastic leukemia. Blood. 2005; 107(5):2061-9. DOI: 10.1182/blood-2005-07-2853. View

19.

Wandler A, Huang B, Craig J, Hayes K, Yan H, Meyer L . Loss of glucocorticoid receptor expression mediates in vivo dexamethasone resistance in T-cell acute lymphoblastic leukemia. Leukemia. 2020; 34(8):2025-2037. PMC: 7440098. DOI: 10.1038/s41375-020-0748-6. View

20.

Malyukova A, Brown S, Papa R, OBrien R, Giles J, Trahair T . FBXW7 regulates glucocorticoid response in T-cell acute lymphoblastic leukaemia by targeting the glucocorticoid receptor for degradation. Leukemia. 2012; 27(5):1053-62. DOI: 10.1038/leu.2012.361. View