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Tumor cells expressing a fusion protein of MULT1 and Fas are rejected in vivo by apoptosis and NK cell activation

Gene Therapy volume 15, pages 1302–1310 (2008)Cite this article

Abstract

Tumor cells evade immunosurveillance by elements of the innate immune system, such as natural killer (NK) cells, by downregulating or ‘shedding’ certain cell-surface molecules like mouse UL16-binding protein-like transcript 1 (MULT1) that can activate NK cells through NK cell receptors such as NKG2D; they also avoid Fas-mediated apoptosis by downregulating its expression. In the present study we report the design and evaluation of the antitumor activity of a novel fusion protein, MULT1E/FasTI, consisting of the extracellular domain of MULT1 and the transmembrane and intracellular domains of Fas. The fusion construct (pMULT1E/FasTI) was transfected into the mouse pulmonary carcinoma cell line TC-1; and stable cell clones expressing the fusion protein were established. In-vitro cell culture studies demonstrated that the binding of the NKG2D/Fc, a recombinant protein of mouse NK cell receptor, to MULT1E/FasTI expressed on tumor cells was able to elicit apoptosis as assayed by Annexin V–fluorescein isothiocyanate staining and caspase-3 enzyme-linked immunosorbent assay and to activate NKG2D-expressing cells, such as NK cells. In-vivo subcutaneous tumor studies demonstrated that tumor cells expressing MULT1E/FasTI grew significantly slower than cells without the protein. Pulmonary metastasis studies showed that most of the mice completely rejected tumor cells expressing MULT1E/FasTI. This approach may generate a new therapeutic agent for tumor treatment when combined with tumor cell-specific gene delivery vehicles such as oncolytic adenovirus vectors.

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Acknowledgements

We thank Eric Holle and his staff for the professional care of the mice used in this study and Lakendra Workman for her administrative contribution in this study. This study was supported in part by the GHS Oncology Foundation and the New Hope fund.

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Authors and Affiliations

  1. Department of Biological Sciences, Clemson University, Clemson, SC, USA

    H S R Kotturi, M Branham-O'Connor, X Yu, T E Wagner & Y Wei

  2. Oncology Research Institute, Greenville Hospital System, Greenville, SC, USA

    J Li, S L Stickel, X Yu, T E Wagner & Y Wei

Authors
  1. H S R Kotturi
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  2. J Li
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  4. S L Stickel
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  7. Y Wei
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Correspondence to Y Wei.

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Kotturi, H., Li, J., Branham-O'Connor, M. et al. Tumor cells expressing a fusion protein of MULT1 and Fas are rejected in vivo by apoptosis and NK cell activation. Gene Ther 15, 1302–1310 (2008). https://doi.org/10.1038/gt.2008.77

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