Letter | Published:

Leukaemia disease genes: large-scale cloning and pathway predictions

Abstract

Retroviral insertional mutagenesis in BXH2 and AKXD recombinant inbred mice induces a high incidence of myeloid or B- and T-cell leukaemia1,2 and the proviral integration sites in the leukaemias provide powerful genetic tags for disease gene identification. Some of the disease genes identified by proviral tagging are also associated with human disease3,4,5, validating this approach for human disease gene identification. Although many leukaemia disease genes have been identified over the years, many more remain to be cloned6. Here we describe an inverse PCR (IPCR) method for proviral tagging that makes use of automated DNA sequencing and the genetic tools provided by the Mouse Genome Project, which increases the throughput for disease gene identification. We also use this IPCR method to clone and analyse more than 400 proviral integration sites from AKXD and BXH2 leukaemias and, in the process, identify more than 90 candidate disease genes. Some of these genes function in pathways already implicated in leukaemia, whereas others are likely to define new disease pathways. Our studies underscore the power of the mouse as a tool for gene discovery and functional genomics.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    Bedigian, H.G., Johnson, D.A., Jenkins, N.A., Copeland, N.G. & Evans, R. Spontaneous and induced leukemias of myeloid origin in recombinant inbred BXH mice. J. Virol. 51, 586–594 (1984).

  2. 2

    Gilbert, D.J., Neumann, P.E., Taylor, B.A., Jenkins, N.A. & Copeland, N.G. Susceptibility of AKXD recombinant inbred mouse strains to lymphomas. J. Virol. 67, 2083–2090 (1993).

  3. 3

    Ogawa, S. et al. Structurally altered Evi-1 protein generated in the 3q21q26 syndrome. Oncogene 13, 183–191 (1996).

  4. 4

    Copeland, N.G. & Jenkins, N.A. Myeloid leukemia: disease genes and mouse models. in Animal Models of Cancer Predisposition Syndromes (eds Hiai, H. & Hino, O.) 53–63 (Karger, Basel, 1999).

  5. 5

    Roberts, T., Chernova, O. & Cowell, J.K. NB4S, a member of the TBC1 domain family of genes, is truncated as a result of a constitutional t(1;10)(p22;q21) chromosome translocation in a patient with stage 4S neuroblastoma. Hum. Mol. Genet. 7, 1169–1178 (1998).

  6. 6

    Look, A.T. Oncogenic transcription factors in the human acute leukemias. Science 278, 1059–1064 (1997).

  7. 7

    Silver, J. & Keerikatte, V. Novel use of polymerase chain reaction to amplify cellular DNA adjacent to an integrated provirus. J. Virol. 63, 1924–1928 (1989).

  8. 8

    Sorensen, A.B., Duch, M., Jorgensen, P. & Pedersen, F.S. Amplification and sequence analysis of DNA flanking integrated proviruses by a simple two-step polymerase chain reaction method. J. Virol. 67, 7118–7124 (1993).

  9. 9

    Valk, P.J.M., Joosten, M., Vankan, Y., Lowenberg, B. & Delwel, R. A rapid RT-PCR based method to isolate complementary DNA fragments flanking retrovirus integration sites. Nucleic Acids Res. 25, 4419–4421 (1997).

  10. 10

    Nakamura, T., Largaespada, D.A., Shaughnessy, J.D. Jr, Jenkins, N.A. & Copeland, N.G. Cooperative activation of Hoxa and Pbx1-related genes in murine myeloid leukaemias. Nature Genet. 12, 149–153 (1996).

  11. 11

    Shannon, K. The Ras signaling pathway and the molecular basis of myeloid leukemogenesis. Curr. Opin. Hematol. 2, 305–308 (1995).

  12. 12

    Kawasaki, H. et al. A Rap guanine nucleotide exchange factor enriched highly in the basal ganglia. Proc. Natl Acad. Sci. USA 95, 13278–13283 (1998).

  13. 13

    Ebinu, J.O. et al. RasGRP, a Ras guanyl nucleotide-releasing protein with calcium- and diacylglycerol-binding motifs. Science 280, 1082–1086 (1998).

  14. 14

    van de Wetering, M., Oosterwegel, M., van Norren, K. & Clevers, H. Sox-4, an Sry-like HMG box protein, is a transcriptional activator in lymphocytes. EMBO J. 12, 3847–3854 (1993).

  15. 15

    Jonkers, J. & Berns, A. Retroviral insertional mutagenesis as a strategy to identify cancer genes. Biochim. Biophys. Acta 1287, 29–57 (1996).

  16. 16

    Carballo, E., Lai, W.S. & Blackshear, P.J. Feedback inhibition of macrophage tumor necrosis factor-α production by tristetraprolin. Science 281, 1001–1005 (1998).

  17. 17

    Taylor, G.A. et al. A pathogenetic role for TNF α in the syndrome of cachexia, arthritis, and autoimmunity resulting from tristetraprolin (TTP) deficiency. Immunity 4, 445–454 (1996).

  18. 18

    Copeland, N.G. & Jenkins, N.A. Development and applications of a molecular genetic linkage map of the mouse genome. Trends Genet. 7, 113–118 (1991).

  19. 19

    Ward, Y. et al. Control of MAP kinase activation by the mitogen-induced threonine/tyrosine phosphatase PAC1. Nature 367, 651–654 (1994).

  20. 20

    Pear, W.S. et al. Exclusive development of T cell neoplasms in mice transplanted with bone marrow expressing activated Notch alleles. J. Exp. Med. 183, 2283–2291 (1996).

  21. 21

    Rohn, J.L., Lauring, A.S., Linenberger, M.L. & Overbaugh, J. Transduction of Notch2 in feline leukemia virus-induced thymic lymphoma. J. Virol. 70, 8071–8080 (1996).

  22. 22

    Ellisen, L.W. et al. TAN-1, the human homolog of the Drosophila notch gene, is broken by chromosomal translocations in T lymphoblastic neoplasms. Cell 66, 649–661 (1991).

  23. 23

    Vitols, S., Gahrton, G., Bjorkholm, M. & Peterson, C. Hypocholesterolaemia in malignancy due to elevated low-density lipoprotein-receptor activity in tumour cells: evidence from studies in patients with leukaemia. Lancet 2, 1150–1154 (1985).

  24. 24

    Cable, S. et al. Peroxisomes in human colon carcinomas. A cytochemical and biochemical study. Virchows Arch. B Cell Pathol. Incl. Mol. Pathol. 62, 221–226 (1992).

  25. 25

    Ericsson, J., Usheva, A. & Edwards, P.A. YY1 is a negative regulator of transcription of three sterol regulatory element-binding protein-responsive genes. J. Biol. Chem. 274, 14508–14513 (1999).

  26. 26

    Austen, M., Cerni, C., Luscher-Firzlaff, J.M. & Luscher, B. YY1 can inhibit c-Myc function through a mechanism requiring DNA binding of YY1 but neither its transactivation domain nor direct interaction with c-Myc. Oncogene 17, 511–520 (1998).

  27. 27

    Jenkins, N.A., Copeland, N.G., Taylor, B.A., Bedigian, H.G. & Lee, B.K. Ecotropic murine leukemia virus DNA content of normal and lymphomatous tissues of BXH-2 recombinant inbred mice. J. Virol. 42, 379–388 (1982).

  28. 28

    Feinberg, A.P. & Vogelstein, B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132, 6–13 (1983).

  29. 29

    Mucenski, M.L., Taylor, B.A., Jenkins, N.A. & Copeland, N.G. AKXD recombinant inbred strains: models for studying the molecular genetic basis of murine lymphomas. Mol. Cell. Biol. 6, 4236–4243 (1986).

  30. 30

    Luo, G., Ivics, Z., Izsvak, Z. & Bradley, A. Chromosomal transposition of a Tc1/mariner-like element in mouse embryonic stem cells. Proc. Natl Acad. Sci. USA 95, 10769–10773 (1998).

Download references

Acknowledgements

We thank N. O'Sullivan for technical assistance; D. Gilbert for help with the IB mapping; and H.C. Morse and M.C. Dean for helpful comments. This research was sponsored in part by the National Cancer Institute, DHHS, under contract with ABL.

Author information

Correspondence to Neal G. Copeland.

Rights and permissions

Reprints and Permissions

About this article

Further reading

Figure 1: Schematic representation of the IPCR proviral tagging method.