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An antibody inhibitor of the LMO2-protein complex blocks its normal and tumorigenic functions

Oncogene volume 27, pages 4962–4968 (2008)Cite this article

Abstract

The LIM-domain protein LMO2 is a T-cell oncogenic protein first recognized by gene activation through chromosomal translocations, but it is also responsible for leukaemias arising as secondary, adverse effects in an X-SCID gene therapy trial. There are no specific reagents currently available to analyse the LMO2 multiprotein complex or to combat LMO2-dependent leukaemias. Accordingly, we have isolated an anti-LMO2 single chain Fv antibody fragment to determine if intracellular interference with LMO2-protein complexes can avert LMO2-dependent functions in normal and cancer settings. The anti-LMO2 single chain Fv, obtained using Intracellular Antibody Capture (IAC) technology, is specific for LMO2 among the LIM-only protein family and binds LMO2 through the third and fourth LIM fingers. Using vector-mediated expression of anti-LMO2 scFv, we show inhibition of Lmo2-dependent erythropoiesis but not endothelial development. We also demonstrate inhibition of Lmo2-dependent leukaemia in a mouse T-cell tumourigenesis transplantation assay with retroviral-mediated expression of anti-LMO2 scFv. Our studies establish that interference with the LMO2 multiprotein complex inhibits both normal and tumourigenic roles. The antibody fragment is a tool for dissecting LMO2 function in haematopoiesis and leukaemia and is a lead for development of therapeutics against LMO2-dependent T-ALL.

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References

  • Alderton G . (2007). Intra-fering with interactions. Nat Rev Cancer 7: 570–571.

    Article  CAS  Google Scholar 

  • Bai S, Thomas C, Rawat A, Ahsan F . (2006). Recent progress in dendrimer-based nanocarriers. Crit Rev Ther Drug Carrier Syst 23: 437–495.

    Article  CAS  PubMed  Google Scholar 

  • Fizzotti M, Chen EY, Link MP, Carroll AJ, Foroni L, Rabbitts TH et al. (1994). Simultaneous expression of RBTN2 and BCR-ABL oncogenes in a T-ALL with a t(11;14)(p13;q11) and a late appearing Philadelphia chromosome. Leukaemia 8: 1124–1130.

    CAS  Google Scholar 

  • Forster A, Pannell R, Drynan L, Cano F, Chan N, Codrington R et al. (2005). Chromosomal translocation engineering to recapitulate primary events of human cancer. Cold Spring Harb Symp Quant Biol 70: 275–282.

    Article  CAS  PubMed  Google Scholar 

  • Grutz G, Forster A, Rabbitts TH . (1998). Identification of the LMO4 gene encoding an interaction partner of the LIM-binding protein LDB1/NLI1: a candidate for displacement by LMO proteins in T cell acute leukaemia. Oncogene 17: 2799–2803.

    Article  CAS  PubMed  Google Scholar 

  • Hacein-Bey-Abina S, von Kalle C, Schmidt M, McCormack MP, Wulffraat N, Leboulch P et al. (2003). LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science 302: 415–419.

    Article  CAS  PubMed  Google Scholar 

  • Holt LJ, Bussow K, Walter G, Tomlinson IM . (2000). By-passing selection: direct screening for antibody-antigen interactions using protein arrays. Nucleic Acids Res 28: E72.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Karlsson O, Thor S, Norberg T, Ohlsson H, Edlund T . (1990). Insulin gene enhancer binding protein Isl-1 is a member of a novel class of proteins containing both a homeo- and a cys-his domain. Nature 344: 879–882.

    Article  CAS  PubMed  Google Scholar 

  • Kenny DA, Jurata LW, Saga Y, Gill GN . (1998). Identification and characterization of LMO4, an LMO gene with a novel pattern of expression during embryogenesis. Proc Natl Acad Sci USA 95: 11257–11262.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Larson R, Fisch P, Larson T, Lavenir I, Langford T, King G et al. (1994). T cell tumours with disparate phenotypes develop with long latency in mice transgenic for rbtn2. Oncogene 9: 3675–3681.

    CAS  PubMed  Google Scholar 

  • Larson RC, Lavenir I, Larson TA, Baer R, Warren AJ, Wadman I et al. (1996). Protein dimerisation between Lmo2 (Rbtn2) and Tal1 alters thymocyte development and potentiates T cell tumorigenesis in transgenic mice. EMBO J 15: 1021–1027.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Larson RC, Osada H, Larson TA, Lavenir I, Rabbitts TH . (1995). The oncogenic LIM protein Rbtn2 causes thymic developmental aberrations that precede malignancy in transgenic mice. Oncogene 11: 853–862.

    CAS  PubMed  Google Scholar 

  • Nam CH, Rabbitts TH . (2006). The role of LMO2 in development and in T cell leukemia after chromosomal translocation or retroviral insertion. Mol Ther 13: 15–25.

    Article  CAS  PubMed  Google Scholar 

  • Neale GA, Rehg JE, Goorha RM . (1995). Ectopic expression of rhombotin-2 causes selective expansion of the thymus and T-cell tumours in transgenic mice. Blood 86: 3060–3071.

    CAS  PubMed  Google Scholar 

  • Peer D, Karp JM, Hong S, Farokhzad OC, Margalit R, Langer R . (2007). Nanocarriers as an emerging platform for cancer therapy. Nat Nanotechnol 2: 751–760.

    Article  CAS  PubMed  Google Scholar 

  • Racevskis J, Dill A, Sparano JA, Ruan H . (1999). Molecular cloning of LMO4, a new human LIM domain gene. Bioch Biophys Acta 1445: 148–153.

    CAS  Google Scholar 

  • Sheets MD, Amersdorfer P, Finnern R, Sargent P, Lindqvist E, Schier R et al. (1998). Efficient construction of a large nonimmune phage antibody library: the production of high-affinity human single-chain antibodies to protein antigens. Proc Natl Acad Sci USA 95: 6157–6162.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Spanopoulou E, Roman CA, Corcoran LM, Schlissel MS, Silver DP, Nemazee D et al. (1994). Functional immunoglobulin transgenes guide ordered B-cell differentiation in Rag-1 deficient mice. Genes Dev 8: 1030–1035.

    Article  CAS  PubMed  Google Scholar 

  • Tanaka T, Lobato MN, Rabbitts TH . (2003). Single domain intracellular antibodies: a minimal fragment for direct in vivo selection of antigen-specific intrabodies. J Mol Biol 331: 1109–1120.

    Article  CAS  PubMed  Google Scholar 

  • Tanaka T, Rabbitts TH . (2003). Intrabodies based on intracellular capture frameworks that bind the RAS protein with high affinity and impair oncogenic transformation. EMBO J 22: 1025–1035.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tanaka T, Williams RL, Rabbitts TH . (2007). Tumour prevention by a single antibody domain inhibiting binding of signal transduction molecules to activated RAS. EMBO J 26: 3250–3259.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tse E, Lobato MN, Forster A, Tanaka T, Chung GTY, Rabbitts TH . (2002). Intracellular antibody capture technology: application to selection of single chain Fv recognising the BCR-ABL oncogenic protein. J Mol Biol 317: 85–94.

    Article  CAS  PubMed  Google Scholar 

  • Van Parijs L, Refaeli Y, Lord JD, Nelson BH, Abbas AK, Baltimore D . (1999). Uncoupling IL-2 signals that regulate T cell proliferation, survival, and Fas-mediated activation-induced cell death. Immunity 11: 281–288.

    Article  CAS  PubMed  Google Scholar 

  • Wadman IA, Osada H, Grutz GG, Agulnick AD, Westphal H, Forster A et al. (1997). The LIM-only protein Lmo2 is a bridging molecule assembling an erythroid, DNA-binding complex which includes the TAL1, E47, GATA-1 and Ldb1/NLI proteins. EMBO J 16: 3145–3157.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Warren AJ, Colledge WH, Carlton MBL, Evans MJ, Smith AJH, Rabbitts TH . (1994). The oncogenic cysteine-rich LIM domain protein rbtn2 is essential for erythroid development. Cell 78: 45–58.

    Article  CAS  PubMed  Google Scholar 

  • Yamada Y, Warren AW, Dobson C, Forster A, Pannell R, Rabbitts TH . (1998). The T cell leukaemia LIM protein Lmo2 is necessary for adult mouse haematopoiesis. Proc Natl Acad Sci USA 95: 3890–3895.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by the Medical Research Council. CHN was supported by a fellowship from the Lady Tata Memorial Trust. We would like to thank Dr T Kitamura (University of Tokyo) for PlatE packaging cells and Dr D Baltimore (California Institute of Technology) for pMIG vector.

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Author notes

  1. C-H Nam

    Present address: 3Current address: Dr C-H Nam, Korea Institute of Science and Technology Europe, Forschungsgesellschaft mbH, Stuhlsatzenhausweg 97, Saarbrücken D-66123, Germany.,

  2. A Appert

    Present address: 4Current address: Dr Alex Appert, Level 3 lab block, Division of Molecular Histopathology, Department of Pathology, Addenbrooke's Hospital, Hills Road, Cambridge.,

Authors and Affiliations

  1. MRC Laboratory of Molecular Biology, Cambridge, UK

    C-H Nam, M N Lobato, A Appert, L F Drynan & T H Rabbitts

  2. Leeds Institute of Molecular Medicine, St James's University Hospital, Leeds, UK

    T Tanaka & T H Rabbitts

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  1. C-H Nam
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  2. M N Lobato
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  4. L F Drynan
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Correspondence to T H Rabbitts.

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The authors have no conflicting financial interests.

Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Nam, CH., Lobato, M., Appert, A. et al. An antibody inhibitor of the LMO2-protein complex blocks its normal and tumorigenic functions. Oncogene 27, 4962–4968 (2008). https://doi.org/10.1038/onc.2008.130

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