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Chronic Myeloproliferative Neoplasias

Inhibition of Grb2 expression demonstrates an important role in BCR–ABL-mediated MAPK activation and transformation of primary human hematopoietic cells

Leukemia volume 25, pages 305–312 (2011)Cite this article

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Abstract

Chronic myeloid leukemia (CML) results from the expression of the BCR/ABL oncogene in a primitive hematopoietic cell. However, BCR/ABL-activated signaling mechanisms are dependent on the cellular context in which it is expressed, and mechanisms underlying primitive human hematopoietic cell transformation by BCR–ABL are not well understood. Our previous studies have shown that BCR/ABL-Y177 has an essential role in Ras activation and human hematopoietic progenitor transformation in CML. The adapter protein growth factor receptor-binding protein-2 (Grb2) can bind phosphorylated BCR/ABL-Y177, induce Grb2-SoS complex formation and activate Ras signaling. We investigated the role of Grb2 in CML progenitor transformation by cotransducing human CD34+ cells with lentivirus vectors expressing short hairpin RNA to Grb2 and retrovirus vectors expressing BCR/ABL. We show that Grb2 knockdown significantly inhibits proliferation and survival of BCR–ABL-expressing CD34+ cells, but not control CD34+ cells. Grb2 knockdown reduced mitogen-activated protein kinase (MAPK) activity in BCR–ABL-expressing hematopoietic cells. We conclude that inhibition of Grb2 expression demonstrates an important role in BCR–ABL-mediated MAPK activation and transformation of primary human hematopoietic cells.These results support further investigation of downstream effectors of Grb2-mediated signals and targeting of Grb2 interactions in the treatment of CML.

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Acknowledgements

This study is supported by the NIH grant R01 HL77847 and R01CA095684 to Ravi Bhatia. We acknowledge the support of the COHNMC Analytical Cytometry and Sequencing cores. We are grateful to StemCyte for their generous gift of cord blood samples.

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Authors and Affiliations

  1. Division of Hematopoietic Stem Cell and Leukemia Research, Department of Hematology and Hematopoietic Cell Transplantation, City of Hope National Medical Center, Duarte, CA, USA

    H Modi, L Li, S Chu & R Bhatia

  2. Department of Molecular and Cellular Biology, City of Hope National Medical Center, Duarte, CA, USA

    J Rossi

  3. Department of Virology Research, City of Hope National Medical Center, Duarte, CA, USA

    J-K Yee

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  1. H Modi
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  2. L Li
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Correspondence to R Bhatia.

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Modi, H., Li, L., Chu, S. et al. Inhibition of Grb2 expression demonstrates an important role in BCR–ABL-mediated MAPK activation and transformation of primary human hematopoietic cells. Leukemia 25, 305–312 (2011). https://doi.org/10.1038/leu.2010.257

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