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Cutting off the power: inhibition of leukemia cell growth by pausing basal ATP release and P2X receptor signaling?

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Abstract

T cells respond to antigen stimulation with the rapid release of cellular ATP, which stimulates an autocrine feedback mechanism that regulates calcium influx through P2X receptors. This autocrine purinergic feedback mechanism plays an essential role in the activation of T cells resulting in cell proliferation and clonal expansion. We recently reported that increases in mitochondrial ATP production drive this stimulation-induced purinergic signaling mechanism but that low-level mitochondrial ATP production fuels basal T cell functions required to maintain vigilance of unstimulated T cells. Here we studied whether defects in these purinergic signaling mechanisms are involved in the unwanted proliferation of leukemia T cells. We found that acute leukemia T cells (Jurkat) possess a larger number and more active mitochondria than their healthy counterparts. Jurkat cells have higher intracellular ATP concentrations and generat more extracellular ATP than unstimulated T cells from healthy donors. As a result, increased purinergic signaling through P2X1 and P2X7 receptors elevates baseline levels of cytosolic Ca2+ in Jurkat cells. We found that pharmacological inhibition of this basal purinergic signaling mechanism decreases mitochondrial activity, Ca2+ signaling, and cell proliferation. Similar results were seen in the leukemic cell lines THP-1, U-937, and HL-60. Combined treatment with inhibitors of P2X1 or P2X7 receptors and the chemotherapeutic agent 6-mercaptopurine completely blocked Jurkat cell proliferation. Our results demonstrate that increased mitochondrial metabolism promotes autocrine purinergic signaling and uncontrolled proliferation of leukemia cells. These findings suggest that deranged purinergic signaling can result in T cell malignancy and that therapeutic targeting aimed at purinergic signaling is a potential strategy to combat T cell leukemia.

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Acknowledgments

This work was funded in part by grants from the National Institutes of Health, GM-51477, GM-60475, AI-080582, and T32GM103702 (W.G.J.), and from the German Research Foundation (DFG), LE-3209/1-1 (C.L.). We thank Drs. Yasutaka Kurishita and Itaru Hamachi for kindly providing the fluorescent ATP probe 2-2Zn(II).

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Authors and Affiliations

  1. Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, Boston, MA, 02215, USA

    Carola Ledderose, Tobias Woehrle, Stephan Ledderose, Katharina Strasser, Richard Seist, Yi Bao, Jingping Zhang & Wolfgang G. Junger

  2. Ludwig Boltzmann Institute for Traumatology, 1200, Vienna, Austria

    Wolfgang G. Junger

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  1. Carola Ledderose
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  2. Tobias Woehrle
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Correspondence to Wolfgang G. Junger.

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All procedures performed in this study involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Supplemental Fig. 1

Staining of mitochondria in Jurkat cells with MitoTracker Green does not depend on mitochondrial membrane potential. a–c Jurkat cells were treated with CCCP (1 μM) or HBSS (control) for 10 min, loaded with MitoTracker Green AM (50 nM) or TMRE (100 nM) for 15 min and analyzed with a flow cytometer. Representative histograms are shown in panels a and b and cumulative results of n = 3 experiments (mean ± SD) are shown in panel c. MTG, MitoTracker Green; MFI, mean fluorescence; n.s., non-significant; ***p < 0.001, t test. (GIF 35 kb)

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Ledderose, C., Woehrle, T., Ledderose, S. et al. Cutting off the power: inhibition of leukemia cell growth by pausing basal ATP release and P2X receptor signaling?. Purinergic Signalling 12, 439–451 (2016). https://doi.org/10.1007/s11302-016-9510-y

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