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Phosphoproteomic analysis reveals hyperactivation of mTOR/STAT3 and LCK/Calcineurin axes in pediatric early T-cell precursor ALL

Leukemia volume 31, pages 1007–1011 (2017)Cite this article

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Early T-cell Precursor Acute Lymphoblastic Leukemia (ETP-ALL), first described in 2009, accounts for about the 10–15% of T-ALL in children.1 Normal ETP cells, also known as double-negative 1 (DN1), the most primitive progenitor T cells in the thymus, retain the ability to differentiate in both T lymphocytes and myeloid cells but not in B lymphocytes.2 ETP-ALL blasts display unique immunophenotype features with lack of T-lineage markers CD1a and CD8, weak or absent CD5, and expression of at least one myeloid or hematopoietic stem cell marker such as CD13, CD33, CD34, CD11b, CD65, HLADR or CD117.1 This disease was initially associated with a very poor prognosis,1, 3 but recent studies report that with intensified protocols, as indicated after a poor early response to induction treatment, the ETP-ALL outcome is not significantly worse than in the other T-ALL subtypes.4, 5, 6 The consolidation phase IB of contemporary AIEOP-BFM protocols, based on cyclophosphamide, 6-mercaptopurine and conventional doses of Ara-C, was effective in clearing disease in most patients,6 supporting the hypothesis that ETP-ALL patients might benefit from some elements of acute myeloid leukemia therapy, as suggested by the similarity to acute myeloid leukemia stem cell subtype gene expression profiling.2, 6 Whole-genome sequencing studies revealed that ETP-ALL patients often display recurrent activating mutations, targeting genes involved in the regulation of cytokine receptors and RAS signaling, inactivating lesions of genes implicated in hematopoietic development, and mutations affecting epigenetic regulation.2 On the other hand, although hyperactivation of the JAK/STAT pathway was observed2, 7, 8 with particular high levels of phospho-STAT1, phospho-STAT3 and phospho-STAT5, proteomic aberrancies of ETP-ALL cells are still poorly understood.

To address this question, we analyzed bone marrow aspirates of 78 AIEOP (Italian Association of Pediatric Hematology and Oncology) T-ALL patients at diagnosis (16 ETP-ALL and 62 other T-ALLs of which 22 early-T, 31 thymic and 9 mature T-ALL defined as described in Supplementary Table S1, collected in the period September 2009–April 2014) through Reverse Phase Protein Arrays (RPPA). The whole blood blast percentage for all samples was between 78 and 97%. Protein lysates were freshly made from bone marrow mononuclear cells separated by Ficoll-Hypaque technique. RPPA analysis was performed as previously described,9 and the activation and/or expression of 50 different proteins was studied (Supplementary Table S2) with a particular focus on AKT/mTOR, JAK/STAT, RAS/MAPK and TCR signaling pathways. Statistical analyses were performed using Prism (GraphPad Software, Inc., La Jolla, CA, USA) and identification of differentially activated or expressed proteins in ETP-ALL patients was obtained through Mann–Whitney test. The activation status of selected pathways was evaluated through Global test analysis10 performed with R (www.r-project.org).

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References

  1. Coustan-Smith E, Mullighan CG, Onciu M, Behm FG, Raimondi SC, Pei D et al. Early T-cell precursor leukaemia: a subtype of very high-risk acute lymphoblastic leukaemia. Lancet Oncol 2009; 10: 147–156.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Zhang J, Ding L, Holmfeldt L, Wu G, Heatley SL, Payne-Turner D et al. The genetic basis of early T-cell precursor acute lymphoblastic leukaemia. Nature 2012; 481: 157–163.

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Ma M, Wang X, Tang J, Xue H, Chen J, Pan C et al. Early T-cell precursor leukemia: a subtype of high risk childhood acute lymphoblastic leukemia. Front Med 2012; 6: 416–420.

    Article  PubMed  Google Scholar 

  4. Patrick K, Wade R, Goulden N, Mitchell C, Moorman AV, Rowntree C et al. Outcome for children and young people with Early T-cell precursor acute lymphoblastic leukaemia treated on a contemporary protocol, UKALL 2003. Br J Haematol 2014; 166: 421–424.

    Article  CAS  PubMed  Google Scholar 

  5. Wood BL, Winter SS, Dunsmore KP, Devidas M, Chen S, Asselin B et al. T-lymphoblastic leukemia (T-ALL) shows excellent outcome, lack of significance of the early thymic precursor (ETP) immunophenotype, and validation of the prognostic value of end-induction minimal residual disease (MRD) in Children’s Oncology Group (COG) Study AALL0434. Blood 2014; 124: 1.

    Article  Google Scholar 

  6. Conter V, Valsecchi MG, Buldini B, Parasole R, Locatelli F, Colombini A et al. Early T-cell precursor acute lymphoblastic leukemia in children treated in AIEOP centres with AIEOP-BFM protocols: a retrospective analysis. Lancet Haematol 2016; 3: e80–e86.

    Article  PubMed  Google Scholar 

  7. Maude SL, Dolai S, Delgado-Martin C, Vincent T, Robbins A, Selvanathan A et al. Efficacy of JAK/STAT pathway inhibition in murine xenograft models of early T-cell precursor (ETP) acute lymphoblastic leukemia. Blood 2015; 125: 1759–1767.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Goossens S, Radaelli E, Blanchet O, Durinck K, Van der Meulen J, Peirs S et al. ZEB2 drives immature T-cell lymphoblastic leukaemia development via enhanced tumour-initiating potential and IL-7 receptor signalling. Nat Commun 2015; 6: 5794.

    Article  CAS  PubMed  Google Scholar 

  9. Milani G, Rebora P, Accordi B, Galla L, Bresolin S, Cazzaniga G et al. Low PKCα expression within the MRD-HR stratum defines a new subgroup of childhood T-ALL with very poor outcome. Oncotarget 2014; 5: 5234–5245.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Goeman JJ, van de Geer SA, de Kort F, van Houwelingen HC . A global test for groups of genes: testing association with a clinical outcome. Bioinformatics 2004; 20: 93–99.

    Article  CAS  PubMed  Google Scholar 

  11. Medyouf H, Alcalde H, Berthier C, Guillemin MC, dos Santos NR, Janin A et al. Targeting calcineurin activation as a therapeutic strategy for T-cell acute lymphoblastic leukemia. Nat Med 2007; 13: 736–741.

    Article  CAS  PubMed  Google Scholar 

  12. Copp J, Manning G, Hunter T . TORC-specific phosphorylation of mammalian target of rapamycin (mTOR): phospho-Ser2481 is a marker for intact mTOR signaling complex 2. Cancer Res 2009; 69: 1821–1827.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Lonetti A, Cappellini A, Spartà AM, Chiarini F, Buontempo F, Evangelisti C et al. PI3K pan-inhibition impairs more efficiently proliferation and survival of T-cell acute lymphoblastic leukemia cell lines when compared to isoform-selective PI3K inhibitors. Oncotarget 2015; 6: 10399–10414.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Ren W, Yin J, Duan J, Liu G, Tan B, Yang G et al. mTORC1 signaling and IL-17 expression: defining pathways and possible therapeutic targets. Eur J Immunol 2016; 46: 291–299.

    Article  CAS  PubMed  Google Scholar 

  15. Zhou J, Wulfkuhle J, Zhang H, Gu P, Yang Y, Deng J et al. Activation of the PTEN/mTOR/STAT3 pathway in breast cancer stem-like cells is required for viability and maintenance. Proc Natl Acad Sci USA 2007; 104: 16158–16163.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Ma J, Meng Y, Kwiatkowski DJ, Chen X, Peng H, Sun Q et al. Mammalian target of rapamycin regulates murine and human cell differentiation through STAT3/p63/jagged/notch cascade. J Clin Invest 2010; 120: 103–114.

    Article  CAS  PubMed  Google Scholar 

  17. Shen YA, Wang CY, Chuang HY, Hwang JJ, Chi WH, Shu CH et al. D44 and CD24 coordinate the reprogramming of nasopharyngeal carcinoma cells towards a cancer stem cell phenotype through STAT3 activation. Oncotarget 2016; 7: 58351–58366.

    PubMed  PubMed Central  Google Scholar 

  18. Crombet O, Lastrapes K, Zieske A, Morales-Arias J . Complete morphologic and molecular remission after introduction of dasatinib in the treatment of a pediatric patient with t-cell acute lymphoblastic leukemia and ABL1 amplification. Pediatr Blood Cancer 2012; 59: 333–334.

    Article  PubMed  Google Scholar 

  19. De Keersmaecker K, Porcu M, Cox L, Girardi T, Vandepoel R, de Beeck JO et al. NUP214-ABL1-mediated cell proliferation in T-cell acute lymphoblastic leukemia is dependent on the LCK kinase and various interacting proteins. Haematologica 2014; 99: 85–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Batista A, Barata JT, Raderschall E, Sallan SE, Carlesso N, Nadler LM et al. Targeting of active mTOR inhibits primary leukemia T cells and synergizes with cytotoxic drugs and signaling inhibitors. Exp Hematol 2011; 39: 457–472.e3.

    Article  CAS  PubMed  Google Scholar 

  21. Zhang C, Ryu YK, Chen TZ, Hall CP, Webster DR, Kang MH . Synergistic activity of rapamycin and dexamethasone in vitro and in vivo in acute lymphoblastic leukemia via cell-cycle arrest and apoptosis. Leuk Res 2012; 36: 342–349.

    Article  CAS  PubMed  Google Scholar 

  22. Daver N, Boumber Y, Kantarjian H, Ravandi F, Cortes J, Rytting ME et al. A Phase I/II study of the mTOR inhibitor everolimus in combination with hyperCVAD chemotherapy in patients with relapsed/refractory acute lymphoblastic leukemia. Clin Cancer Res 2015; 21: 2704–2714.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by grants from Fondazione CARIPARO-Istituto di Ricerca Pediatrica Città della Speranza (n.13/05) and AIRC (IG 2016, Id.19186); VS is supported by a fellowship from Fondazione Italiana per la Ricerca sul Cancro (n.16616). We thank B Michielotto, S Disarò and Dr P Scarparo (University of Padova) for technical assistance, and Dr E Giarin (Istituto di Ricerca Pediatrica 'Città della Speranza') for help in patients data management.

Author contributions

VS, VL and BA conceived the experiments; VS and VL performed the experiments; VS and BA analyzed the data; BA and SB performed statistical analyses; BB provided the patients data and classification; MP, FG, NA, CP and GC performed mutational screening; BA, VS, VC and SM interpreted the data; BA and SM wrote the manuscript; VC, GB and BA supervised the research.

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Author notes

  1. N Andriano

    Present address: 5Present address: Department of Pediatric Hemato-Oncology, Azienda Policlinico-Ospedale Vittorio Emanuele, Catania, Italy.,

  2. G Basso and B Accordi: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Woman’s and Child’s Health, University of Padova, Padova, Italy

    V Serafin, V Lissandron, B Buldini, S Bresolin, M Paganin, F Grillo, G Basso & B Accordi

  2. Department of Pediatrics, Centro Ricerca Tettamanti, University of Milano-Bicocca, Fondazione MBBM, Monza, Italy

    N Andriano, C Palmi & G Cazzaniga

  3. Department of Surgery, Medicine, Dentistry and Morphology, University of Modena and Reggio Emilia, Modena, Italy

    S Marmiroli

  4. Department of Pediatrics, Center of Hemato-Oncology, University of Milano-Bicocca, Fondazione MBBM, Ospedale San Gerardo, Monza, Italy

    V Conter

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Correspondence to B Accordi.

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Serafin, V., Lissandron, V., Buldini, B. et al. Phosphoproteomic analysis reveals hyperactivation of mTOR/STAT3 and LCK/Calcineurin axes in pediatric early T-cell precursor ALL. Leukemia 31, 1007–1011 (2017). https://doi.org/10.1038/leu.2017.13

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