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Lentiviral marking of patient-derived acute lymphoblastic leukaemic cells allows in vivo tracking of disease progression

Leukemia volume 27, pages 718–721 (2013)Cite this article

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A Corrigendum to this article was published on 07 August 2013

Biological studies of acute leukaemias are hampered by the technical difficulties involved in maintaining the viability and proliferation of patient-derived blasts in vitro. This is particularly true for studies trying to elucidate the nature and biology of leukaemia-propagating cells, the definition of which relies on functional assays of self-renewal/clonal expansion. Furthermore, the lack of critical niche interactions raises concerns over the validity of biological data generated purely in an in vitro setting, for example, when using derived cell line approaches1 or patient-derived cells with compromised viability. Developments in immunocompromised mouse models, notably NOD/scid and more recently the NOD/LtSz-scid IL-2 Rγ null (NSG)2 and similar mice, have provided an opportunity to study leukaemia-propagating cells in a bone marrow, albeit xenogeneic, microenvironment.3, 4, 5, 6 Alongside this, there is a need to develop techniques to allow the labelling and tracking of leukaemic blasts in such xenotransplantation assays.

Multicolour flow cytometry demonstrated the unbiased transduction of all identified populations defined by the precursor B-cell surface markers CD19, CD20 and CD34 (Figure 1b). During the development of the protocol, a total of 10 transductions (7 primary samples, 3 ‘primografted’ samples, which had been passaged through mice) were performed on five different patient-derived specimens with a mean transduction efficiency of 21% (range 2–42%) (Supplementary Table S1). The two final transductions were performed using viral stocks, which had previously been titrated using the B-lymphoblastic leukaemia cell line SEM7 and shown in that system to have 5.6 × 107–1.2 × 108 transducing units/ml. These experiments showed the relative transduction efficiency in L826 to be 0.5–3.4% and L839 to be 1.6–11%, compared with that in SEM.

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References

  1. Meacham CE, Ho EE, Dubrovsky E, Gertler FB, Hemann MT . In vivo RNAi screening identifies regulators of actin dynamics as key determinants of lymphoma progression. Nat Genet 2009; 41: 1133–1137.

    Article  CAS  Google Scholar 

  2. Shultz LD, Lyons BL, Burzenski LM, Gott B, Chen X, Chaleff S et al. Human lymphoid and myeloid cell development in NOD/LtSz-scid IL2R gamma null mice engrafted with mobilized human hemopoietic stem cells. J Immunol 2005; 174: 6477–6489.

    Article  CAS  Google Scholar 

  3. Hong D, Gupta R, Ancliff P, Atzberger A, Brown J, Soneji S et al. Initiating and cancer-propagating cells in TEL-AML1-associated childhood leukemia. Science 2008; 319: 336–339.

    Article  CAS  Google Scholar 

  4. Kong Y, Yoshida S, Saito Y, Doi T, Nagatoshi Y, Fukata M et al. CD34+CD38+CD19+ as well as CD34+CD38-CD19+ cells are leukemia-initiating cells with self-renewal capacity in human B-precursor ALL. Leukemia 2008; 22: 1207–1213.

    Article  CAS  Google Scholar 

  5. le Viseur C, Hotfilder M, Bomken S, Wilson K, Rottgers S, Schrauder A et al. In childhood acute lymphoblastic leukemia, blasts at different stages of immunophenotypic maturation have stem cell properties. Cancer Cell 2008; 14: 47–58.

    Article  CAS  Google Scholar 

  6. Martinez Soria N, Tussiwand R, Ziegler P, Manz MG, Heidenreich O . Transient depletion of RUNX1/RUNX1T1 by RNA interference delays tumour formation in vivo. Leukemia 2009; 23: 188–190.

    Article  CAS  Google Scholar 

  7. Greil J, Gramatzki M, Burger R, Marschalek R, Peltner M, Trautmann U et al. The acute lymphoblastic leukaemia cell line SEM with t(4;11) chromosomal rearrangement is biphenotypic and responsive to interleukin-7. Br J Haematol 1994; 86: 275–283.

    Article  CAS  Google Scholar 

  8. Kustikova OS, Modlich U, Fehse B . Retroviral insertion site analysis in dominant haematopoietic clones. Methods Mol Biol 2009; 506: 373–390.

    Article  CAS  Google Scholar 

  9. Schmidt M, Hoffmann G, Wissler M, Lemke N, Mussig A, Glimm H et al. Detection and direct genomic sequencing of multiple rare unknown flanking DNA in highly complex samples. Hum Gene Ther 2001; 12: 743–749.

    Article  CAS  Google Scholar 

  10. Bonnet D, Dick JE . Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med 1997; 3: 730–737.

    Article  CAS  Google Scholar 

  11. Hope KJ, Jin L, Dick JE . Acute myeloid leukemia originates from a hierarchy of leukemic stem cell classes that differ in self-renewal capacity. Nat Immunol 2004; 5: 738–743.

    Article  CAS  Google Scholar 

  12. Lapidot T, Sirard C, Vormoor J, Murdoch B, Hoang T, Caceres-Cortes J et al. A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Nature 1994; 367: 645–648.

    Article  CAS  Google Scholar 

  13. Rehe K, Wilson K, Bomken S, McNeill H, Stanulla M, Den Boer ML et al. In acute lymphoblastic leukaemia, stemness is frequent and ubiquitous. Blood 2010; 116, : Abstract 92.

  14. Morisot S, Wayne AS, Bohana-Kashtan O, Kaplan IM, Gocke CD, Hildreth R et al. High frequencies of leukemia stem cells in poor-outcome childhood precursor-B acute lymphoblastic leukemias. Leukemia 2010; 24: 1859–1866.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Christopher Baum and Olga Kustikova for their help with the ligation-mediated PCR and Martin Stanulla and Martin Schrappe for the provision of sample L4951. SB is funded by a Medical Research Council (MRC) Clinical Research Training Fellowship. The IVIS Spectrum was funded by a grant from the Wellcome Trust, grant 087961. Additional funding came from Leukaemia Lymphoma Research (LLR) and core infrastructure support from the North of England Children′s Cancer Research Fund (NECCR).

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Authors and Affiliations

  1. Newcastle Cancer Centre, Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, UK

    S Bomken, L Buechler, K Rehe, F Ponthan, A Elder, H Blair, C M Bacon, J Vormoor & O Heidenreich

  2. Department of Paediatric and Adolescent Haematology and Oncology, Great North Children′s Hospital, Newcastle upon Tyne Hospitals NHS Foundation Trust, Newcastle upon Tyne, UK

    S Bomken, K Rehe & J Vormoor

  3. Department of Cellular Pathology, Newcastle upon Tyne Hospitals NHS Foundation Trust, Newcastle upon Tyne, UK

    C M Bacon

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Correspondence to O Heidenreich.

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Bomken, S., Buechler, L., Rehe, K. et al. Lentiviral marking of patient-derived acute lymphoblastic leukaemic cells allows in vivo tracking of disease progression. Leukemia 27, 718–721 (2013). https://doi.org/10.1038/leu.2012.206

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