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Targeting PYK2 mediates microenvironment-specific cell death in multiple myeloma

Oncogene volume 35, pages 2723–2734 (2016)Cite this article

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Abstract

Multiple myeloma (MM) remains an incurable malignancy due, in part, to the influence of the bone marrow microenvironment on survival and drug response. Identification of microenvironment-specific survival signaling determinants is critical for the rational design of therapy and elimination of MM. Previously, we have shown that collaborative signaling between β1 integrin-mediated adhesion to fibronectin and interleukin-6 confers a more malignant phenotype via amplification of signal transducer and activator of transcription 3 (STAT3) activation. Further characterization of the events modulated under these conditions with quantitative phosphotyrosine profiling identified 193 differentially phosphorylated peptides. Seventy-seven phosphorylations were upregulated upon adhesion, including PYK2/FAK2, Paxillin, CASL and p130CAS consistent with focal adhesion (FA) formation. We hypothesized that the collaborative signaling between β1 integrin and gp130 (IL-6 beta receptor, IL-6 signal transducer) was mediated by FA formation and proline-rich tyrosine kinase 2 (PYK2) activity. Both pharmacological and molecular targeting of PYK2 attenuated the amplification of STAT3 phosphorylation under co-stimulatory conditions. Co-culture of MM cells with patient bone marrow stromal cells (BMSC) showed similar β1 integrin-specific enhancement of PYK2 and STAT3 signaling. Molecular and pharmacological targeting of PYK2 specifically induced cell death and reduced clonogenic growth in BMSC-adherent myeloma cell lines, aldehyde dehydrogenase-positive MM cancer stem cells and patient specimens. Finally, PYK2 inhibition similarly attenuated MM progression in vivo. These data identify a novel PYK2-mediated survival pathway in MM cells and MM cancer stem cells within the context of microenvironmental cues, providing preclinical support for the use of the clinical stage FAK/PYK2 inhibitors for treatment of MM, especially in a minimal residual disease setting.

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Abbreviations

ASO:

anti-sense oligonucleotide

BMSC:

bone marrow stromal cell

ERK:

extracellular signal-regulated kinase

FAK:

focal adhesion kinase

FCM:

flow cytometry method

FERM:

4.1 protein, ezrin, radixin, moesin domain

FN:

fibronectin

gp130:

IL-6 beta receptor, IL-6 signal transducer

IL-6:

interleukin-6

JAK:

Janus kinase

MRD:

minimal residual disease

PYK2:

proline-rich tyrosine kinase 2

STAT:

signal transducer and activator of transcription

Sus:

suspension

TME:

tumor microenvironment.

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Acknowledgements

We thank Jianguo Tao and William S Dalton for helpful discussions and critical reading of the manuscript. This work was supported by Florida Department of Health Bankhead-Coley Team Science Program grant 2BT03 (to KSH and LHA). The Moffitt Proteomics Facility is supported by the US Army Medical Research and Materiel Command under Award W81XWH-08-2-0101 for a National Functional Genomics Center, the National Cancer Institute under Award P30-CA076292 as a Cancer Center Support Grant and the Moffitt Foundation. Patient specimens were obtained from the Total Cancer Care program at Moffitt Cancer Center. Patient specimen collection, phosphoproteome mapping and flow cytometry were performed by the Translational Research, Proteomics and Flow Cytometry Core facilities at Moffitt Cancer Center. Antisense oligonucleotides were provided by Isis Pharmaceuticals (Carlsbad, CA, USA). The focal adhesion kinase inhibitors VS-6062,VS-6063 (defactinib), and VS-4718 were provided by Verastem, Inc. (Needham, MA, USA).

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Authors and Affiliations

  1. Chemical Biology and Molecular Medicine Program, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA

    M B Meads, L Mathews, J Gemmer, L Nong, I Rosado-Lopez, T Nguyen & K H Shain

  2. Department of Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA

    M B Meads, L Mathews, J Gemmer, L Nong, I Rosado-Lopez, T Nguyen & K H Shain

  3. Proteomics Core Facility, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA

    B Fang & J M Koomen

  4. Verastem, Inc., Needham, Massachusetts, USA

    J E Ring & J A Pachter

  5. Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    W Matsui

  6. ISIS Pharmaceuticals, Carlsbad, CA, USA

    A R MacLeod

  7. Department of Molecular Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA

    L A Hazlehurst & J M Koomen

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Correspondence to K H Shain.

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Meads, M., Fang, B., Mathews, L. et al. Targeting PYK2 mediates microenvironment-specific cell death in multiple myeloma. Oncogene 35, 2723–2734 (2016). https://doi.org/10.1038/onc.2015.334

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