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MECHANISMS OF RESISTANCE

Sustained activation of non-canonical NF-κB signalling drives glycolytic reprogramming in doxorubicin-resistant DLBCL

Abstract

DLBCL is the most common lymphoma with high tumor heterogeneity. Treatment refractoriness and relapse from R-CHOP therapy in patients remain a clinical problem. Activation of the non-canonical NF-κB pathway is associated with R-CHOP resistance. However, downstream targets of non-canonical NF-κB mediating R-CHOP-induced resistance remains uncharacterized. Here, we identify the common mechanisms underlying both intrinsic and acquired resistance that are induced by doxorubicin, the main cytotoxic component of R-CHOP. We performed global transcriptomic analysis of (1) a panel of resistant versus sensitive and (2) isogenic acquired doxorubicin-resistant DLBCL cell lines following short and chronic exposure to doxorubicin respectively. Doxorubicin-induced stress in resistant cells activates a distinct transcriptional signature that is enriched in metabolic reprogramming and oncogenic signalling. Selective and sustained activation of non-canonical NF-κB signalling in these resistant cells exacerbated their survival by augmenting glycolysis. In response to doxorubicin, p52-RelB complexes transcriptionally activated multiple glycolytic regulators with prognostic significance through increased recruitment at their gene promoters. Targeting p52-RelB and their targets in resistant cells increased doxorubicin sensitivity in vitro and in vivo. Collectively, our study uncovered novel molecular drivers of doxorubicin-induced resistance that are regulated by non-canonical NF-κB pathway. We reveal new avenues of therapeutic targeting for R-CHOP-treated refractory/relapsed DLBCL patients.

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Fig. 1: DLBCL cell lines display differential responsiveness to doxorubicin-induced cytotoxicity which correlates with the apoptotic response to doxorubicin-induced DDR.
Fig. 2: Resistant DLBCL cells display a distinct doxorubicin-induced transcriptomic signature that predicts pan cancer doxorubicin sensitivity.
Fig. 3: Periodic treatment of sensitive DLBCL cells to increasing doses of doxorubicin results in the acquisition of doxorubicin resistance and reduction in doxorubicin-induced DDR.
Fig. 4: Dox-induced transcriptomes of acquired and primary Dox-resistant DLBCL cells are similarly enriched in metabolic reprogramming and oncogenic processes.
Fig. 5: Sustained activation of non-canonical NF-κB pathway promotes survival in Dox-resistant cells via enhanced glycolysis.
Fig. 6: Dox-induced recruitment of p52-RelB to the promoters of clinically important metabolic regulators results in their increased expression in Dox-resistant cells.
Fig. 7: Inhibition of p52-RelB regulated targets reverses doxorubicin resistance and attenuates doxorubicin-induced glycolysis in DLBCL cells.
Fig. 8: Inhibition of non-canonical NF-κB signalling abolishes doxorubicin resistance in DLBCL in vivo.

Data availability

The RNA-seq datasets generated during and/or analysed during the current study had been deposited in the GEO (Gene Expression Omnibus) database under the accession number GSE215900.

References

  1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021;71:209–49.

    Article  Google Scholar 

  2. Susanibar-Adaniya S, Barta SK. 2021 Update on Diffuse large B cell lymphoma: A review of current data and potential applications on risk stratification and management. Am J Hematol. 2021;96:617–29.

    Article  Google Scholar 

  3. Wu JQ, Song YP, Su LP, Zhang MZ, Li W, Hu Y, et al. Three-year Follow-up on the Safety and Effectiveness of Rituximab Plus Chemotherapy as First-Line Treatment of Diffuse Large B-Cell Lymphoma and Follicular Lymphoma in Real-World Clinical Settings in China: A Prospective, Multicenter, Noninterventional Study. Chin Med J (Engl). 2018;131:1767–75.

    Article  Google Scholar 

  4. He MY, Kridel R. Treatment resistance in diffuse large B-cell lymphoma. Leukemia. 2021;35:2151–65.

    Article  Google Scholar 

  5. Palmer AC, Chidley C, Sorger PK. A curative combination cancer therapy achieves high fractional cell killing through low cross-resistance and drug additivity. Elife 2019;8:e50036.

    Article  Google Scholar 

  6. Nowakowski GS, Czuczman MSABC, GCB, and Double-Hit Diffuse Large B-Cell Lymphoma: Does Subtype Make a Difference in Therapy Selection? Am Soc Clin Oncol Educ Book 2015: e449–57.

  7. Erttmann R, Erb N, Steinhoff A, Landbeck G. Pharmacokinetics of doxorubicin in man: dose and schedule dependence. J Cancer Res Clin Oncol. 1988;114:509–13.

    Article  CAS  Google Scholar 

  8. Yang W, Soares J, Greninger P, Edelman EJ, Lightfoot H, Forbes S, et al. Genomics of Drug Sensitivity in Cancer (GDSC): a resource for therapeutic biomarker discovery in cancer cells. Nucl Acids Res. 2013;41:D955–961.

    Article  CAS  Google Scholar 

  9. Bonner WM, Redon CE, Dickey JS, Nakamura AJ, Sedelnikova OA, Solier S, et al. GammaH2AX and cancer. Nat Rev Cancer. 2008;8:957–67.

    Article  CAS  Google Scholar 

  10. Bieging KT, Mello SS, Attardi LD. Unravelling mechanisms of p53-mediated tumour suppression. Nat Rev Cancer. 2014;14:359–70.

    Article  CAS  Google Scholar 

  11. Chen X, Chen S, Yu D. Metabolic Reprogramming of Chemoresistant Cancer Cells and the Potential Significance of Metabolic Regulation in the Reversal of Cancer Chemoresistance. Metabolites 2020;10:289.

    Article  CAS  Google Scholar 

  12. Zhong Z, Virshup DM. Wnt Signaling and Drug Resistance in Cancer. Mol Pharm. 2020;97:72–89.

    Article  CAS  Google Scholar 

  13. Flahaut M, Meier R, Coulon A, Nardou KA, Niggli FK, Martinet D, et al. The Wnt receptor FZD1 mediates chemoresistance in neuroblastoma through activation of the Wnt/beta-catenin pathway. Oncogene. 2009;28:2245–56.

    Article  CAS  Google Scholar 

  14. Yuniati L, Scheijen B, van der Meer LT, van Leeuwen FN. Tumor suppressors BTG1 and BTG2: Beyond growth control. J Cell Physiol. 2019;234:5379–89.

    Article  CAS  Google Scholar 

  15. Tamura RE, de Vasconcellos JF, Sarkar D, Libermann TA, Fisher PB, Zerbini LF. GADD45 proteins: central players in tumorigenesis. Curr Mol Med. 2012;12:634–51.

    Article  CAS  Google Scholar 

  16. Bensaad K, Tsuruta A, Selak MA, Vidal MN, Nakano K, Bartrons R, et al. TIGAR, a p53-inducible regulator of glycolysis and apoptosis. Cell. 2006;126:107–20.

    Article  CAS  Google Scholar 

  17. Xu H, Yan M, Patra J, Natrajan R, Yan Y, Swagemakers S, et al. Enhanced RAD21 cohesin expression confers poor prognosis and resistance to chemotherapy in high grade luminal, basal and HER2 breast cancers. Breast Cancer Res. 2011;13:R9.

    Article  CAS  Google Scholar 

  18. Yu J, Zhou J, Xu F, Bai W, Zhang W. High expression of Aurora-B is correlated with poor prognosis and drug resistance in non-small cell lung cancer. Int J Biol Markers. 2018;33:215–21.

    Article  CAS  Google Scholar 

  19. Zhu Y, Li K, Zhang J, Wang L, Sheng L, Yan L. Inhibition of CDK1 Reverses the Resistance of 5-Fu in Colorectal Cancer. Cancer Manag Res. 2020;12:11271–83.

    Article  CAS  Google Scholar 

  20. Barretina J, Caponigro G, Stransky N, Venkatesan K, Margolin AA, Kim S, et al. Addendum: The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature. 2019;565:E5–E6.

    Article  CAS  Google Scholar 

  21. Zhou Y, Tozzi F, Chen J, Fan F, Xia L, Wang J, et al. Intracellular ATP levels are a pivotal determinant of chemoresistance in colon cancer cells. Cancer Res. 2012;72:304–14.

    Article  CAS  Google Scholar 

  22. Vander Heiden MG, Cantley LC, Thompson CB. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science. 2009;324:1029–33.

    Article  Google Scholar 

  23. Xu Y, Fang F, St Clair DK, Sompol P, Josson S, St Clair WH. SN52, a novel nuclear factor-kappaB inhibitor, blocks nuclear import of RelB:p52 dimer and sensitizes prostate cancer cells to ionizing radiation. Mol Cancer Ther. 2008;7:2367–76.

    Article  CAS  Google Scholar 

  24. Zhao B, Barrera LA, Ersing I, Willox B, Schmidt SC, Greenfeld H, et al. The NF-kappaB genomic landscape in lymphoblastoid B cells. Cell Rep. 2014;8:1595–606.

    Article  CAS  Google Scholar 

  25. de Oliveira KA, Kaergel E, Heinig M, Fontaine JF, Patone G, Muro EM, et al. A roadmap of constitutive NF-kappaB activity in Hodgkin lymphoma: Dominant roles of p50 and p52 revealed by genome-wide analyses. Genome Med. 2016;8:28.

    Article  Google Scholar 

  26. Li J, He Y, Tan Z, Lu J, Li L, Song X, et al. Wild-type IDH2 promotes the Warburg effect and tumor growth through HIF1alpha in lung cancer. Theranostics. 2018;8:4050–61.

    Article  CAS  Google Scholar 

  27. Zhao J, Li J, Fan TWM, Hou SX. Glycolytic reprogramming through PCK2 regulates tumor initiation of prostate cancer cells. Oncotarget. 2017;8:83602–18.

    Article  Google Scholar 

  28. Jiang Y, He R, Jiang Y, Liu D, Tao L, Yang M, et al. Transcription factor NFAT5 contributes to the glycolytic phenotype rewiring and pancreatic cancer progression via transcription of PGK1. Cell Death Dis. 2019;10:948.

    Article  CAS  Google Scholar 

  29. Saumet A, Vetter G, Bouttier M, Antoine E, Roubert C, Orsetti B, et al. Estrogen and retinoic acid antagonistically regulate several microRNA genes to control aerobic glycolysis in breast cancer cells. Mol Biosyst. 2012;8:3242–53.

    Article  CAS  Google Scholar 

  30. Liu W, Sun Y, Ge W, Zhang F, Gan L, Zhu Y, et al. DIA-Based Proteomics Identifies IDH2 as a Targetable Regulator of Acquired Drug Resistance in Chronic Myeloid Leukemia. Mol Cell Proteom. 2022;21:100187.

    Article  CAS  Google Scholar 

  31. Vaeth M, Feske S. NFAT control of immune function: New Frontiers for an Abiding Trooper. F1000Res. 2018;7:260.

    Article  Google Scholar 

  32. Bruce JIE. Metabolic regulation of the PMCA: Role in cell death and survival. Cell Calcium. 2018;69:28–36.

    Article  CAS  Google Scholar 

  33. Lovas A, Weidemann A, Albrecht D, Wiechert L, Weih D, Weih F. p100 Deficiency is insufficient for full activation of the alternative NF-kappaB pathway: TNF cooperates with p52-RelB in target gene transcription. PLoS One. 2012;7:e42741.

    Article  CAS  Google Scholar 

  34. Yang Y, Kelly P, Shaffer AL 3rd, Schmitz R, Yoo HM, Liu X, et al. Targeting Non-proteolytic Protein Ubiquitination for the Treatment of Diffuse Large B Cell Lymphoma. Cancer Cell. 2016;29:494–507.

    Article  CAS  Google Scholar 

  35. Simpson L, Ansell SM, Colgan JP, Habermann TM, Inwards DJ, Ristow KM, et al. Effectiveness of second line salvage chemotherapy with ifosfamide, carboplatin, and etoposide in patients with relapsed diffuse large B-cell lymphoma not responding to cis-platinum, cytosine arabinoside, and dexamethasone. Leuk Lymphoma. 2007;48:1332–7.

    Article  CAS  Google Scholar 

  36. Sobrevilla-Calvo P, Vargas-Hernandez L, Cortes-Padilla D, Labardini-Mendez JR, Oñate-Ocaña LF. Dexamethasone, etoposide and cisplatin (DEP) as second line chemotherapy in patients with diffuse large B cell lymphoma (DLBCL). J Clin Oncol. 2005;23:6723.

    Article  Google Scholar 

  37. Wang X, Yan J, Shen B, Wei G. Integrated Chromatin Accessibility and Transcriptome Landscapes of Doxorubicin-Resistant Breast Cancer Cells. Front Cell Dev Biol. 2021;9:708066.

    Article  Google Scholar 

  38. Park HY, Lee SB, Yoo HY, Kim SJ, Kim WS, Kim JI, et al. Whole-exome and transcriptome sequencing of refractory diffuse large B-cell lymphoma. Oncotarget. 2016;7:86433–45.

    Article  Google Scholar 

  39. Pham LV, Tamayo AT, Li C, Lee J, Fayad L, Ford RJ. Chemo-Resistance in Diffuse Large Cell Lymphoma: Novel Drug Combinations Targeting NFAT/NF-Kb Growth/Survival/Chemo-Resistance Signaling Pathways in Validated Novel Experimental Systems. Blood. 2011;118:1428.

    Article  Google Scholar 

  40. Eluard B, Nuan-Aliman S, Faumont N, Collares D, Bordereaux D, Montagne A, et al. The alternative RelB NF-kappaB subunit is a novel critical player in diffuse large B-cell lymphoma. Blood. 2022;139:384–98.

    Article  CAS  Google Scholar 

  41. McGuirk S, Audet-Delage Y, Annis MG, Xue Y, Vernier M, Zhao, K et al. Resistance to different anthracycline chemotherapeutics elicits distinct and actionable primary metabolic dependencies in breast cancer. Elife 2021;10:e65150.

    Article  CAS  Google Scholar 

  42. Nuan-Aliman S, Bordereaux D, Thieblemont C, Baud V. The Alternative RelB NF-kB Subunit Exerts a Critical Survival Function upon Metabolic Stress in Diffuse Large B-Cell Lymphoma-Derived Cells. Biomedicines 2022;10:348.

    Article  CAS  Google Scholar 

  43. Gorini S, De Angelis A, Berrino L, Malara N, Rosano G, Ferraro E. Chemotherapeutic Drugs and Mitochondrial Dysfunction: Focus on Doxorubicin, Trastuzumab, and Sunitinib. Oxid Med Cell Longev. 2018;2018:7582730.

    Article  Google Scholar 

  44. Jing Z, Gao J, Li J, Niu F, Tian L, Nan P, et al. Acetylation-induced PCK isoenzyme transition promotes metabolic adaption of liver cancer to systemic therapy. Cancer Lett. 2021;519:46–62.

    Article  CAS  Google Scholar 

  45. Cole AJ, Iyengar M, Panesso-Gomez S, O’Hayer P, Chan D, Delgoffe GM, et al. NFATC4 promotes quiescence and chemotherapy resistance in ovarian cancer. JCI Insight. 2020;5:e131486.

    Article  Google Scholar 

  46. de Sousa EMF, Vermeulen L. Wnt Signaling in Cancer Stem Cell Biology. Cancers (Basel) 2016; 8:60.

  47. El-Sahli S, Xie Y, Wang L, Liu S. Wnt Signaling in Cancer Metabolism and Immunity. Cancers (Basel) 2019;11:904.

    Article  CAS  Google Scholar 

  48. Du Q, Geller DA. Cross-Regulation Between Wnt and NF-kappaB Signaling. Pathw Immunopathol Dis Ther. 2010;1:155–81.

    Google Scholar 

  49. Schwitalla S, Fingerle AA, Cammareri P, Nebelsiek T, Goktuna SI, Ziegler PK, et al. Intestinal tumorigenesis initiated by dedifferentiation and acquisition of stem-cell-like properties. Cell. 2013;152:25–38.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Hui Fen Kerry May Lim and Dachuan Huang from National Cancer Centre, Singapore for establishing the PDX model. This study is funded by the National Research Foundation (NRF) Singapore, under its Singapore NRF Fellowship (NRF-NRFF2018-04). In addition, we thank the Nanyang Assistant Professorship (NAP) Start-up-grant to Y.L. lab and National Medical Research Council (NMRC-OFLCG-18May0028), Tanoto Foundation and Ling Foundation for their support.

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YL conceptualized and supervised the study. YL and SKL planned and devised the experiments. SKL, SL and AB performed all molecular and cell biology experiments. CCP and VV performed all bioinformatics analyses and data visualization. STL and CKO contributed the PDX model and BHP performed the PDX ex vivo experiments. ST and SKL conducted the tumor xenograft studies. SKL analysed the data and JQL reviewed all statistical analyses. ADJ provided feedback on the study design. SKL wrote the manuscript and YL edited it.

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Correspondence to Yinghui Li.

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Lim, S.K., Peng, C.C., Low, S. et al. Sustained activation of non-canonical NF-κB signalling drives glycolytic reprogramming in doxorubicin-resistant DLBCL. Leukemia 37, 441–452 (2023). https://doi.org/10.1038/s41375-022-01769-w

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