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Abnormality of c-kit oncoprotein in certain patients with chronic myelogenous leukemia – potential clinical significance

Leukemia volume 16, pages 170–177 (2002)Cite this article

Abstract

Chronic myelogenous leukemia (CML) is characterized by the Philadelphia (Ph) chromosome and bcr/abl gene rearrangement which occurs in pluripotent hematopoietic progenitor cells expressing the c-kit receptor tyrosine kinase (KIT). To elucidate the biological properties of KIT in CML leukemogenesis, we performed analysis of alterations of the c-kit gene and functional analysis of altered KIT proteins. Gene alterations in the c-kit juxtamembrane domain of 80 CML cases were analyzed by reverse transcriptase and polymerase chain reaction-single strand conformation polymorphism (RT-PCR-SSCP). One case had an abnormality at codon 564 (AAT→AAG, Asn→Lys), and six cases had the same base abnormality at codon 541 (ATG→CTG, Met→Leu) in the juxtamembrane domain. Because the change from Met to Leu at codon 541 was a conservative one which was also observed in the normal population and normal tissues of CML patients, it probably represents a polymorphic variation. Although samples of hair roots and leukemic cells from the chronic phase of one CML patient showed no abnormality, an abnormality at codon 541 (ATG→CTG, Met→Leu) was found only at blastic crisis (BC) of this case. In the case with the abnormality at codon 564, the mutation was detected only in a sample of leukemic cells collected at BC. To examine the biological consequence and biological significance of these abnormalities, murine KITL540 and KITK563 expression vectors were introduced into interleukin-3 (IL-3)-dependent murine Ba/F3 cells to study their state of tyrosine phosphorylation and their growth rate. Ba/F3 cells expressing KITWT, KITL540 and KITK563 showed dose-dependent tyrosine phosphorylation after treatment with increasing concentrations of recombinant mouse stem cell factor (rmSCF). The cells expressing KITL540 and KITK563 were found to have greater tyrosine phosphorylation than cells expressing KITWT at 0.1 and 1.0 ng/ml of rmSCF. The Ba/F3 cells expressing KITK563 proliferated in response to 0.1 and 1.0 ng/ml of rmSCF as well as IL-3. The Ba/F3 cells expressing KITL540showed a relatively higher proliferative response to 0.1 ng/ml of rmSCF than the response of cells expressing KITWT. These mutations and in vitro functional analyses raise the possibility that the KIT abnormalities influence the white blood cell counts (P < 0.05) and survival (P < 0.04) of CML patients.

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References

  1. Groffen J, Stephenson JR, Heisterkamp N, de Klein A, Bartram CR, Grosveld G . Philadelphia chromosomal breakpoints are clustered within a limited region, bcr, on chromosome 22 Cell 1984 36: 93–99

    Article  CAS  PubMed  Google Scholar 

  2. Kantarjian HM, Smith TL, McCredie KB, Keating MJ, Walters RS, Talpaz M, Hester JP, Bligham G, Gehan E, Freireich EJ . Chronic myelogenous leukemia: a multivariate analysis of the associations of patient characteristics and therapy with survival Blood 1985 66: 1326–1335

    CAS  PubMed  Google Scholar 

  3. Shtivelman E, Lifshiftz B, Gale RP, Canaani E . Fused transcripts of abl and bcr genes in chronic myelogenous leukemia Nature 1985 315: 550–554

    Article  CAS  PubMed  Google Scholar 

  4. Inokuchi K, Inoue T, Tojo A, Futaki M, Miyake K, Yamada T, Tanabe Y, Ohki I, Dan K, Ozawa K, Asano S, Nomura T . A possible correlation between the type of bcr-abl hybrid messenger RNA and platelet count in Philadelphia-positive chronic myelogenous leukemia Blood 1991 78: 3125–3127

    CAS  PubMed  Google Scholar 

  5. Yamaguchi H, Inokuchi K, Shinohara T, Dan K . Extramedullary presentation of chronic myelogenous leukemia with p190 BCR/ABL transcripts Cancer Genet Cytogenet 1998 102: 74–77

    Article  CAS  PubMed  Google Scholar 

  6. Yamagata T, Mitani K, Kanda Y, Yazaki Y, Hirai H . Elevated platelet count features the variant type of BCR/ABL junction in chronic myelogenous leukemia Br J Haematol 1996 94: 370–372

    Article  CAS  PubMed  Google Scholar 

  7. Mills KI, Birnie GD . Does the breakpoint within the major breakpoint cluster region (M-bcr) influence the duration of the chronic phase in chronic myeloid leukemia? An analytical comparison of current literature Blood 1991 78: 1155–1161

    CAS  PubMed  Google Scholar 

  8. Shtalrid M, Talpaz M, Kurzrock R, Kantarjian H, Trujillo J, Gutterman J, Galina Y, Mark B . Analysis of breakpoints within the bcr gene and their correlation with clinical course of Philadelphia-positive chronic myelogenous leukemia Blood 1988 72: 485–490

    CAS  PubMed  Google Scholar 

  9. Faderl S, Talpaz M, Estrov Z, O'Brien S, Kurzrock R, Kantarjian HM . The biology of chronic myeloid leukemia N Engl J Med 1999 341: 164–172

    Article  CAS  PubMed  Google Scholar 

  10. Melo JV, Myint H, Galton DAG, Goldman JM . P190 BCR-ABL chronic myeloid leukemia: the missing link with chronic myelomonocytic leukemia? Leukemia 1994 8: 208–211

    CAS  PubMed  Google Scholar 

  11. Ogawa M, Matsuzaki Y, Nishikawa S, Hayashi S-I, Kunisada T, Sudo T, Kina H, Nakauchi H, Nishikawa S-I . Expression and function of c-kit in hematopoietic progenitor cells J Exp Med 1991 174: 63–71

    Article  CAS  PubMed  Google Scholar 

  12. Bernstein ID, Andrew RG, Zsebo KM . Recombinant human stem cell factor enhances formation of colonies by CD34+ and CD34+lin− cells, and the generation of colony-forming cell progeny from CD34+lin− cells cultured with interleukin-3, granulocyte colony-stimulating factor, or granulocyte–macrophage colony-stimulating factor Blood 1991 77: 2316–2321

    CAS  PubMed  Google Scholar 

  13. Palacios R, Steinmetz M . IL3-dependent mouse clone that express B-220 surface antigen, contain Ig genes in germ-line configration, and generate B lymphocytes in vivo Cell 1985 41: 727–734

    Article  CAS  PubMed  Google Scholar 

  14. Tanosaki S, Inokuchi K, Shimada T, Dan K . Relation between microsatellite instability and N-ras mutation and duration of disease free survival in patients with acute leukemia Cancer 1998 83: 475–481

    Article  CAS  PubMed  Google Scholar 

  15. Abo J, Inokuchi K, Dan K, Nomura T . p53 and N-ras mutations in two new leukemia cell lines established from a patient with multilineage CD7-positive acute leukemia Blood 1993 82: 2829–2836

    CAS  PubMed  Google Scholar 

  16. Inokuchi K, Fukaki M, Hanawa H, Tanosaki S, Yamaguchi H, Iwakiri R, Nomura T, Dan K . Heterogenous expression of bcr-abl fusion mRNA in a patient with Philadelphia-chromosome-positive acute lymphoblastic leukemia Br J Haematol 1997 97: 837–840

    Article  CAS  PubMed  Google Scholar 

  17. Furitsu T, Tsujimura T, Yono T, Ikeda H, Kitayama H, Koshimizu U, Sugahara H, Butterfield JH, Ashman LK, Kanayama Y, Matsuzawa Y, Kitamura Y, Kanakura Y . Identification of mutations in the coding sequence of the proto-oncogene c-kit in a human mast cell leukemia cell line causing ligand-dependent activation of c-kit J Clin Invest 1993 92: 1736–1744

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Mizushima S, Nagata S . pEF-BOS, a powerful mammalian expression vector Nucleic Acids Res 1990 18: 5322

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Inokuchi K, Miyake K, Takahashi H, Dan K, Nomura T . DCC protein expression in hematopoietic cell populations and its relation to leukemogenesis J Clin Invest 1996 97: 852–857

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Martin-Henao GA, Quiroga R, Sureda A, Garcia S . CD7 expression on CD34+ cells from chronic myeloid leukemia in chronic phase Am J Hematol 1999 61: 178–186

    Article  CAS  PubMed  Google Scholar 

  21. Hirota S, Isozaki K, Moriyama Y, Hashimoto K, Nishida T, Ishiguro S, Kawano K, Hanada M, Kurata A, Takeda M, Tunio GM, Matsuzawa Y, Kanakura Y, Shinomura Y, Kitamura Y . Gain-of-function mutations of c-kit in human gastrointestinal stromal tumors Science 1998 279: 577–580

    Article  CAS  PubMed  Google Scholar 

  22. Tsujimura T, Furitsu T, Morimoto M, Isozaki K, Nomura S, Matsuzawa Y, Kitamura Y, Kanakura Y . Ligand-independent activation of c-kit receptor tyrosine kinase in a murine mastocytoma cell line P-815 generated by a point mutation Blood 1994 83: 2619–2626

    CAS  PubMed  Google Scholar 

  23. Sakurai S, Fukasawa T, Chong JM, Tanaka A, Fukayama M . C-kit gene abnormalities in gastrointestinal stromal tumors (tumors of interstitial cells of Cajal) Jpn J Cancer Res 1999 90: 1321–1328

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Tsujimura T, Morimoto M, Hashimoto K, Moriyama Y, Kitayama H, Matsuzawa Y, Kitamura Y, Kanakura Y . Constitutive activation of c-kit in FMA3 murine mastocytoma cells caused by deletion of seven amino acids at the juxtamembrane domain Blood 1996 87: 273–283

    CAS  PubMed  Google Scholar 

  25. Kiyoi H, Naoe T, Nakano Y, Yokota S, Minami S, Miyawaki S, Asou N, Kuriyama K, Jinnai I, Shimazaki C, Akiyama H, Saito K, Oh H, Moyoji T, Omoto E, Saito H, Ohno R, Ueda R . Prognostic implication of FLT3 and N-RAS gene mutations in acute myeloid leukemia Blood 1999 93: 3074–3080

    CAS  PubMed  Google Scholar 

  26. Matthews W, Jordan CT, Wiegand GW, Pardoll D, Lemischka IR . A receptor tyrosine kinase specific to hematopoietic stem and progenitor cell-enriched populations Cell 1991 65: 1143–1152

    Article  CAS  PubMed  Google Scholar 

  27. Hayakawa F, Towatari M, Kiyoi H, Tanimoto M, Kitamura T, Saito H, Naoe T . Tandem-duplicated Flt3 constitutively activates STAT5 and MAP kinase and introduces autonomous cell growth in IL-3-dependent cell lines Oncogene 2000 19: 624–631

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr Y Kanakura for providing full-length murine c-kit expression vectors and Kirin Brewery Company, Ltd for providing rmIL-3 and rmSCF. This work was supported by Grants-in-Aid from the Ministry of Education, Science and Culture of Japan and the Takahashi Foundation.

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  1. Division of Hematology, Department of Internal Medicine, Nippon Medical School, Tokyo, Japan

    K Inokuchi, H Yamaguchi, M Tarusawa, M Futaki, H Hanawa, S Tanosaki & K Dan

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  1. K Inokuchi
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  2. H Yamaguchi
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Inokuchi, K., Yamaguchi, H., Tarusawa, M. et al. Abnormality of c-kit oncoprotein in certain patients with chronic myelogenous leukemia – potential clinical significance. Leukemia 16, 170–177 (2002). https://doi.org/10.1038/sj.leu.2402341

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