ABB  Vol.5 No.2 , January 2014
Juvenile myelo-monocytic leukemia (JMML): No effect of granulocyte monocyte-colony stimulating factor (GM-CSF) on Wilms Tumor gene (WT1) by nested Polymerase Chain Reaction (nPCR) and flow cytometry

This study was to determine whether GM-CSF induced WT1 gene expression and to establish an association with markers of proliferation CD71+CD34+ using nPCR and flow cytometry respectively, in samples obtained from 5 newly diagnosed JMML patients. Overtime (day 0 to day 14) there was an insignificant difference in WT1 gene expression and CD71+CD34+ in JMML samples when compared to peripheral blood of normal volunteers (n = 3). Our study suggests that there is a correlation between WT1 gene expression and cellular proliferation and that GMCSF in vitro does not create a significant difference in JMML samples.

Cite this paper
Khan, S. , Olszewski, M. , Huang, W. and Kletzel, M. (2014) Juvenile myelo-monocytic leukemia (JMML): No effect of granulocyte monocyte-colony stimulating factor (GM-CSF) on Wilms Tumor gene (WT1) by nested Polymerase Chain Reaction (nPCR) and flow cytometry. Advances in Bioscience and Biotechnology, 5, 155-159. doi: 10.4236/abb.2014.52019.
[1]   Grillo, Niemeyer, C.M., Arico, M. and Basso, G. (1997) Chronic myelomonocytic leukemia in childhood: A retrospective analysis of 110 cases. Blood, 89, 3534-3543.

[2]   Emanuel, P.D. (2008) Juvenile myelomonocytic leukemia and chronic myelomonocytic leukemia. Leukemia, 22, 1335-1342.

[3]   Emanuel, P.D., Bates, L.J., Castleberry, R.P., Gualtieri, R.J. and Zuckerman, K.S. (1991) Selective hypersensitivity to granulocyte-macrophage colony-stimulating factor by juvenile myeloid leukemia hematopoietic progenitors. Blood, 77, 925-929.

[4]   de Vries, A.C., Zwaan, C.M. and van den Heuvel-Eibrink, M.M. (2010) Molecular basis of juvenile myelomonocytic leukemia. Hematologica, 95, 179-181.

[5]   Kenichi, K. and Kazuyuki, M. (2008) Recent advances in the pathogenesis and management ofjuvenile myelomonocyticleukaemia. British Journal of Haematology, 141, 567-575.

[6]   Yoshida, N., Yagasaki, H., Xu, Y.Y., et al. (2009) Correlation of clinical features with the mutational status of GMCSF signaling pathway-related genes in juvenile myelomonocytic leukemia. Pediatric Research, 65, 334-340.

[7]   Gualtieri, R.J., Emanuel, P.D., Zuckerman, K.S., et al. (1989) Granulocyte-macrophage colony-stimulating factor is an endogenous regulator of cell proliferation in juvenile chronic myelogenous leukemia. Blood, 74, 2360-2367.

[8]   Scharnhorst, V., van der Eb, A.J. and Jochenmsen, A.G. (2001) WT1 proteins: Functions in growth and differentiation. Gene, 273, 141-161.

[9]   Inoue, K., Sugiyama, H., Ogawa, H., et al. (1994) WT1 as a new prognostic factor and a new marker for the detection of minimal residual disease in acute leukemia. Blood, 84, 3071-3079.

[10]   Chan, R.J., Cooper, T., Kratz, C.P., Weiss, B. and Loh, M.L. (2009) Juvenile myelomonocytic leukemia: A report from the 2nd International JMML symposium. Leukemia Research, 33, 355-362.

[11]   Jacobsohn, D.A., Tse, W.T., Cheleff, S., et al. (2009) High WT1 gene expression before hematopoietic stem cell transplant in children with acute myeloid leukemia predicts poor event-free survival. British Journal of Haematology, 146, 669-674.

[12]   Cilloni, D., Gottardi, E., Messa, F., et al. (2003) Significant correlation between the degree if WT1 expression and the international prognostic scoring system score in patients with myelodysplastic syndromes. Journal of Clinical Oncology, 21, 1988-1995.

[13]   Kletzel, M., Olzewski, M., Huang, W. and Chou, P.M. (2002) Utility of WT1 as a reliable tool for the detection of minimal residual disease in children with leukemia. Pediatric and Developmental Pathology, 5, 269-275.

[14]   Sugiyama, H. (1998) Wilms’ tumor gene (WT1) as a new marker for the detection of minimal residual disease in leukemia. Leuk Lymphoma, 30, 55-61.

[15]   Ostergaard, M., Olesen, L.H., Hasle, H., Kjeldsen, E. and Hokland, P. (2004) WT1 gene expression: An excellent tool for monitoring minimal residual disease in 70% of acute myeloid leukemia patients-results from a singlecenter study. British Journal of Haematology, 125, 590-600.

[16]   Bender, J.G., Unverzagt, K., Walker, D.E., et al. (1994) Phenotypic analysis and characterization of CD34+ cells from normal human bone marrow, cord blood, peripheral blood, and mobilized peripheral blood from patients undergoing autologous stem cell transplantation. Clinical Immunology and Immunopathology, 70, 10-18.

[17]   D’Arena, G., Cascavilla, N., Musto, P., et al. (1996) Flow cytometric characterization of CD34+ hematopoietic progenitor cells in mobilized peripheral blood and bone marrow of cancer patients. Haematologica, 81, 216-223.

[18]   Hoffman, R., Tong, J., Brandt, J., et al. (1993) The in vitro and in vivo effects of stem cell factor on human hematopoiesis. Stem Cells, 2, 76-82.

[19]   Hosen, N., Sonoda, Y., Oji, Y., et al. (2002) Very low frequencies of human normal CD34+ haematopoietic progenitor cells express the wilms’ tumour gene WT1 at levels similar to those in leukaemia cells. British Journal of Haematology, 116, 409-420.

[20]   Campana, D. and Coustan-Smith, E. (2004) Minimal residual disease studies by flow cytometry in acute leukemia. Acta Haematologica, 112, 8-15.

[21]   Olszewski, M., Huang, W., Chou, P.M., Duerst, R. and Kletzel, M. (2005) Wilms’ tumor 1 (WT1) gene in hematopoiesis: A surrogate marker of cell proliferation as a possible mechanism of action? Cytotherapy, 7, 57-61.

[22]   Tamaki, H., Ogawa, H., Ohyashiki, J.H., et al. (2005) The Wilms’ tumor is a good marker for diagnosis of disease progression of myelodysplastic syndrome. Leukemia, 29, 343-345.

[23]   Bader, P., Niemeyer, C., Weber, G., et al. (2004) WT1 gene expression: Useful marker for minimal residual disease in childhood myelodysplastic syndromes and juvenile myelomonocytic leukemia? European Journal of Haematology, 73, 25-28.

[24]   Ogawa, H., Tamaki, H., Ikegame, K., et al. (2003) The usefulness of monitoring WT1 gene transcripts for the prediction and management of relapse following allogeneic stem cell transplantation in acute type leukemia. Blood, 101, 5.

[25]   Qin, Y.Z., Zhu, H.H., Jiang, B. et al. (2009) Expression patterns of WT1 and PRAME in acute myeloid leukemia patients and their usefulness for monitoring minimal residual disease. Leukemia Research, 33, 384-390.

[26]   Phelan, S.A., Lindberg, C. and Call, K.M. (1994) Wilms’ tumor gene, WT1, mRNA, is down-regulated during induction of erythroid and megakaryocytic differentiation of K562 cells. Cell Growth & Differentiation, 5, 677-686.