Precursor-B-cell receptor (pre-BCR) signaling and spleen tyrosine kinase (SYK) recently were introduced as therapeutic targets for patients with B-cell acute lymphoblastic leukemia (B-ALL), but the importance of this pathway in B-ALL subsets and mechanism of downstream signaling have not fully been elucidated. Here, we provide new detailed insight into the mechanism of pre-BCR signaling in B-ALL. We compared the effects of pharmacological and genetic disruption of pre-BCR signaling in vitro and in mouse models for B-ALL, demonstrating exquisite dependency of pre-BCR+ B-ALL, but not other B-ALL subsets, on this signaling pathway. We demonstrate that SYK, PI3K/AKT, FOXO1 and MYC are important downstream mediators of pre-BCR signaling in B-ALL. Furthermore, we define a characteristic immune phenotype and gene expression signature of pre-BCR+ ALL to distinguish them from other B-ALL subsets. These data provide comprehensive new insight into pre-BCR signaling in B-ALL and corroborate pre-BCR signaling and SYK as promising new therapeutic targets in pre-BCR+ B-ALL.
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Roberts KG, Morin RD, Zhang J, Hirst M, Zhao Y, Su X et al. Genetic alterations activating kinase and cytokine receptor signaling in high-risk acute lymphoblastic leukemia. Cancer Cell 2012; 22: 153–166.
Mullighan CG, Goorha S, Radtke I, Miller CB, Coustan-Smith E, Dalton JD et al. Genome-wide analysis of genetic alterations in acute lymphoblastic leukaemia. Nature 2007; 446: 758–764.
Herzog S, Reth M, Jumaa H . Regulation of B-cell proliferation and differentiation by pre-B-cell receptor signalling. Nat Rev Immunol 2009; 9: 195–205.
Mårtensson I-L, Almqvist N, Grimsholm O, Bernardi AI . The pre-B cell receptor checkpoint. FEBS Lett 2010; 584: 2572–2579.
Kitamura D, Roes J, Kühn R, Rajewsky KA . B cell-deficient mouse by targeted disruption of the membrane exon of the immunoglobulin μ chain gene. Nature 1991; 350: 423–426.
Kitamura D, Kudo A, Schaal S, Müller W, Melchers F, Rajewsky K . A critical role of λ5 protein in B cell development. Cell 1992; 69: 823–831.
Bankovich AJ, Raunser S, Juo ZS, Walz T, Davis MM, Garcia KC . Structural insight into Pre-B cell receptor function. Science 2007; 316: 291–294.
Übelhart R, Bach MP, Eschbach C, Wossning T, Reth M, Jumaa H . N-linked glycosylation selectively regulates autonomous precursor BCR function. Nat Immunol 2010; 11: 759–765.
Ohnishi K, Melchers F . The nonimmunoglobulin portion of λ5 mediates cell-autonomous pre-B cell receptor signaling. Nat Immunol 2003; 4: 849–856.
Yasuda T, Sanjo H, Pagès G, Kawano Y, Karasuyama H, Pouysségur J et al. Erk kinases link Pre-B cell receptor signaling to transcriptional events required for early B cell expansion. Immunity 2008; 28: 499–508.
Herzog S, Hug E, Meixlsperger S, Paik J-H, DePinho RA, Reth M et al. SLP-65 regulates immunoglobulin light chain gene recombination through the PI(3)K-PKB-Foxo pathway. Nat Immunol 2008; 9: 623–631.
Eswaran J, Sinclair P, Heidenreich O, Irving J, Russell LJ, Hall A et al. The pre-B-cell receptor checkpoint in acute lymphoblastic leukaemia. Leukemia 2015; 29: 1623–1631.
Chen Z, Shojaee S, Buchner M, Geng H, Lee JW, Klemm L et al. Signalling thresholds and negative B-cell selection in acute lymphoblastic leukaemia. Nature 2015; 521: 357–361.
Geng H, Hurtz C, Lenz KB, Chen Z, Baumjohann D, Thompson S et al. Self-enforcing feedback activation between BCL6 and Pre-B cell receptor signaling defines a distinct subtype of acute lymphoblastic leukemia. Cancer Cell 2015; 27: 409–425.
Duy C, Hurtz C, Shojaee S, Cerchietti L, Geng H, Swaminathan S et al. BCL6 enables Ph+ acute lymphoblastic leukaemia cells to survive BCR-ABL1 kinase inhibition. Nature 2011; 473: 384–388.
Trageser D, Iacobucci I, Nahar R, Duy C, Levetzow G, von, Klemm L et al. Pre–B cell receptor–mediated cell cycle arrest in Philadelphia chromosome–positive acute lymphoblastic leukemia requires IKAROS function. J Exp Med 2009; 206: 1739–1753.
Jumaa H, Bossaller L, Portugal K, Storch B, Lotz M, Flemming A et al. Deficiency of the adaptor SLP-65 in pre-B-cell acute lymphoblastic leukaemia. Nature 2003; 423: 452–456.
Feldhahn N, Klein F, Mooster JL, Hadweh P, Sprangers M, Wartenberg M et al. Mimicry of a constitutively active pre–B cell receptor in acute lymphoblastic leukemia cells. J Exp Med 2005; 201: 1837–1852.
Swaminathan S, Huang C, Geng H, Chen Z, Harvey R, Kang H et al. BACH2 mediates negative selection and p53-dependent tumor suppression at the pre-B cell receptor checkpoint. Nat Med 2013; 19: 1014–1022.
Bicocca VT, Chang BH, Masouleh BK, Muschen M, Loriaux MM, Druker BJ et al. Crosstalk between ROR1 and the Pre-B cellreceptor promotes survival of t(1;19) acute lymphoblastic leukemia. Cancer Cell 2012; 22: 656–667.
Perova T, Grandal I, LMJ Nutter, Papp E, Matei IR, Beyene J et al. Therapeutic potential of spleen tyrosine kinase inhibition for treating high-risk precursor B cell acute lymphoblastic leukemia. Sci Transl Med 2014; 6: 236ra62.
Uckun FM, Qazi S, Cely I, Sahin K, Shahidzadeh A, Ozercan I et al. Nanoscale liposomal formulation of a SYK P-site inhibitor against B-precursor leukemia. Blood 2013; 121: 4348–4354.
Uckun FM, Ek RO, Jan S-T, Chen C-L, Qazi S . Targeting SYK kinase-dependent anti-apoptotic resistance pathway in B-lineage acute lymphoblastic leukaemia (ALL) cells with a potent SYK inhibitory pentapeptide mimic. Br J Haematol 2010; 149: 508–517.
van der Veer A, van der Velden VHJ, Willemse ME, Hoogeveen PG, Petricoin EF, Beverloo HB et al. Interference with pre-B-cell receptor signaling offers a therapeutic option for TCF3-rearranged childhood acute lymphoblastic leukemia. Blood Cancer J 2014; 4: e181.
Clark MR, Mandal M, Ochiai K, Singh H . Orchestrating B cell lymphopoiesis through interplay of IL-7 receptor and pre-B cell receptor signalling. Nat Rev Immunol 2014; 14: 69–80.
Espeli M, Rossi B, Mancini SJC, Roche P, Gauthier L, Schiff C . Initiation of pre-B cell receptor signaling: common and distinctive features in human and mouse. Semin Immunol 2006; 18: 56–66.
Hoellenriegel J, Coffey GP, Sinha U, Pandey A, Sivina M, Ferrajoli A et al. Selective, novel spleen tyrosine kinase (Syk) inhibitors suppress chronic lymphocytic leukemia B-cell activation and migration. Leukemia 2012; 26: 1576–1583.
Ann Ran F, Hsu Patrick D, Wright Jason, Agarwala Vineeta, Scott David A, Zhang Feng . Genome engineering using the CRISPR-Cas9 system. Nat Protoc 2013; 8: 2281–2308.
Nakamura N, Ramaswamy S, Vazquez F, Signoretti S, Loda M, Sellers WR . Forkhead transcription factors are critical effectors of cell death and cell cycle arrest downstream of PTEN. Mol Cell Biol 2000; 20: 8969–8982.
Ma W, Wang M, Wang Z-Q, Sun L, Graber D, Matthews J et al. Effect of long-term storage in TRIzol on microarray-based gene expression profiling. Cancer Epidemiol Biomark Prev 2010; 19: 2445–2452.
Lee S-T, Xiao Y, Muench MO, Xiao J, Fomin ME, Wiencke JK et al. A global DNA methylation and gene expression analysis of early human B-cell development reveals a demethylation signature and transcription factor network. Nucleic Acids Res 2012; 40: 11339–11351.
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.
Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci USA 2005; 102: 15545–15550.
Mootha VK, Lindgren CM, Eriksson K-F, Subramanian A, Sihag S, Lehar J et al. PGC-1α-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes. Nat Genet 2003; 34: 267–273.
Hystad ME, Myklebust JH, Bø TH, Sivertsen EA, Rian E, Forfang L et al. Characterization of early stages of human B cell development by gene expression profiling. J Immunol 2007; 179: 3662–3671.
Hoffmann R, Lottaz C, Kühne T, Rolink A, Melchers F . Neutrality, compensation, and negative selection during evolution of B-cell development transcriptomes. Mol Biol Evol 2007; 24: 2610–2618.
Tsuganezawa K, Kiyokawa N, Matsuo Y, Kitamura F, Toyama-Sorimachi N, Kuida K et al. Flow cytometric diagnosis of the cell lineage and developmental stage of acute lymphoblastic leukemia by novel monoclonal antibodies specific to human Pre–B-cell receptor. Blood 1998; 92: 4317–4324.
Reilly MP, Sinha U, André P, Taylor SM, Pak Y, DeGuzman FR et al. PRT-060318, a novel Syk inhibitor, prevents heparin-induced thrombocytopenia and thrombosis in a transgenic mouse model. Blood 2011; 117: 2241–2246.
Cheng S, Coffey G, Zhang XH, Shaknovich R, Song Z, Lu P et al. SYK inhibition and response prediction in diffuse large B-cell lymphoma. Blood 2011; 118: 6342–6352.
Chen L, Monti S, Juszczynski P, Ouyang J, Chapuy B, Neuberg D et al. SYK inhibition modulates distinct PI3K/AKT- dependent survival pathways and cholesterol biosynthesis in diffuse large B cell lymphomas. Cancer Cell 2013; 23: 826–838.
Coffey G, DeGuzman F, Inagaki M, Pak Y, Delaney SM, Ives D et al. Specific inhibition of spleen tyrosine kinase suppresses leukocyte immune function and inflammation in animal models of rheumatoid arthritis. J Pharmacol Exp Ther 2012; 340: 350–359.
Guo B, Kato RM, Garcia-Lloret M, Wahl MI, Rawlings DJ . Engagement of the human pre-B cell receptor generates a lipid raft–dependent calcium signaling complex. Immunity 2000; 13: 243–253.
Dong X-Y, Chen C, Sun X, Guo P, Vessella RL, Wang R-X et al. FOXO1A is a candidate for the 13q14 tumor suppressor gene inhibiting androgen receptor signaling in prostate cancer. Cancer Res 2006; 66: 6998–7006.
Xie L, Ushmorov A, Leithäuser F, Guan H, Steidl C, Färbinger J et al. FOXO1 is a tumor suppressor in classical Hodgkin lymphoma. Blood 2012; 119: 3503–3511.
Sykes SM, Lane SW, Bullinger L, Kalaitzidis D, Yusuf R, Saez B et al. AKT/FOXO signaling enforces reversible differentiation blockade in myeloid leukemias. Cell 2011; 146: 697–708.
Heng TSP, Painter MW . Immunological Genome Project Consortium. The immunological genome project: networks of gene expression in immune cells. Nat Immunol 2008; 9: 1091–1094.
Gan B, Lim C, Chu G, Hua S, Ding Z, Collins M et al. FoxOs enforce a progression checkpoint to constrain mTORC1-activated renal tumorigenesis. Cancer Cell 2010; 18: 472–484.
Alizadeh AA, Eisen MB, Davis RE, Ma C, Lossos IS, Rosenwald A et al. Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling. Nature 2000; 403: 503–511.
Braselmann S, Taylor V, Zhao H, Wang S, Sylvain C, Baluom M et al. R406, an orally available spleen tyrosine kinase inhibitor blocks Fc receptor signaling and reduces immune complex-mediated inflammation. J Pharmacol Exp Ther 2006; 319: 998–1008.
Davis MI, Hunt JP, Herrgard S, Ciceri P, Wodicka LM, Pallares G et al. Comprehensive analysis of kinase inhibitor selectivity. Nat Biotechnol 2011; 29: 1046–1051.
Roberts KG, Li Y, Payne-Turner D, Harvey RC, Yang Y-L, Pei D et al. Targetable kinase-activating lesions in Ph-like acute lymphoblastic leukemia. N Engl J Med 2014; 371: 1005–1015.
Maude SL, Tasian SK, Vincent T, Hall JW, Sheen C, Roberts KG et al. Targeting JAK1/2 and mTOR in murine xenograft models of Ph-like acute lymphoblastic leukemia. Blood 2012; 120: 3510–3518.
Wossning T, Herzog S, Kohler F, Meixlsperger S, Kulathu Y, Mittler G et al. Deregulated Syk inhibits differentiation and induces growth factor-independent proliferation of pre-B cells. J Exp Med 2006; 203: 2829–2840.
Herman SEM, Gordon AL, Hertlein E, Ramanunni A, Zhang X, Jaglowski S et al. Bruton tyrosine kinase represents a promising therapeutic target for treatment of chronic lymphocytic leukemia and is effectively targeted by PCI-32765. Blood 2011; 117: 6287–6296.
Ponader S, Chen S-S, Buggy JJ, Balakrishnan K, Gandhi V, Wierda WG et al. The Bruton tyrosine kinase inhibitor PCI-32765 thwarts chronic lymphocytic leukemia cell survival and tissue homing in vitro and in vivo. Blood 2012; 119: 1182–1189.
Hunger SP . Chromosomal translocations involving the E2A gene in acute lymphoblastic leukemia: clinical features and molecular pathogenesis. Blood 1996; 87: 1211–1224.
Habib T, Park H, Tsang M, de Alborán IM, Nicks A, Wilson L et al. Myc stimulates B lymphocyte differentiation and amplifies calcium signaling. J Cell Biol 2007; 179: 717–731.
Suzuki H, Terauchi Y, Fujiwara M, Aizawa S, Yazaki Y, Kadowaki T et al. Xid-Like immunodeficiency in mice with disruption of the p85α subunit of phosphoinositide 3-Kinase. Science 1999; 283: 390–392.
Aiba Y, Kameyama M, Yamazaki T, Tedder TF, Kurosaki T . Regulation of B-cell development by BCAP and CD19 through their binding to phosphoinositide 3-kinase. Blood 2008; 111: 1497–1503.
Ramadani F, Bolland DJ, Garcon F, Emery JL, Vanhaesebroeck B, Corcoran AE et al. The PI3K Isoforms p110α and p110δ are essential for Pre–B cell receptor signaling and B cell development. Sci Signal 2010; 3: ra60.
Turner M, Joseph Mee P, Costello PS, Williams O, Price AA, Duddy LP et al. Perinatal lethality and blocked B-cell development in mice lacking the tyrosine kinase Syk. Nature 1995; 378: 298–302.
This work was supported by a Cancer Prevention and Research Institute of Texas (CPRIT) grant (to JAB), a Leukemia & Lymphoma Society Scholar Award in Clinical Research (to JAB), the Else-Kröner Forschungskolleg (FS), German Research Foundation, SFB 1074 (KMD and LHM), ‘Förderkreis für Tumor- und Leukämiekranke Kinder Ulm’ and in part by the MD Anderson Cancer Center Support Grant CA016672. STR DNA fingerprinting was done by the Cancer Center Support Grant-funded Characterized Cell Line core, NCI # CA016672. We are grateful for access to the data assembled by the ImmGen Consortium.
JAB received research funding from Portola Pharmaceuticals and GPC is employee and shareholder of Portola Pharmaceuticals.
This work was presented, in part, in oral sessions at the 2013 annual meeting of the American Society of Hematology (ASH) in New Orleans and at the 2014 annual meeting of the American Society of Hematology (ASH) in San Francisco.
Supplementary Information accompanies this paper on the Leukemia website
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Köhrer, S., Havranek, O., Seyfried, F. et al. Pre-BCR signaling in precursor B-cell acute lymphoblastic leukemia regulates PI3K/AKT, FOXO1 and MYC, and can be targeted by SYK inhibition. Leukemia 30, 1246–1254 (2016) doi:10.1038/leu.2016.9
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