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Radiation-induced autophagy potentiates immunotherapy of cancer via up-regulation of mannose 6-phosphate receptor on tumor cells in mice

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Abstract

There is a significant body of evidence demonstrating that radiation therapy (XRT) enhances the effect of immune therapy. However, the precise mechanisms by which XRT potentiates the immunotherapy of cancer remain elusive. Here, we report that XRT potentiates the effect of immune therapy via induction of autophagy and resultant trafficking of mannose-6-phopsphate receptor (MPR) to the cell surface. Irradiation of different tumor cells caused substantial up-regulation of MPR on the cell surface in vitro and in vivo. Down-regulation of MPR in tumor cells with shRNA completely abrogated the combined effect of XRT and immunotherapy (CTLA4 antibody) in B16F10-bearing mice without changes in the tumor-specific responses of T cells. Radiation-induced MPR up-regulation was the result of redistribution of the receptor to the cell surface. This effect was caused by autophagy with redirection of MPR to autophagosomes in a clathrin-dependent manner. In autophagosomes, MPR lost its natural ligands, which resulted in subsequent trafficking of empty receptor(s) back to the surface. Together, our data demonstrated a novel mechanism by which XRT can enhance the effect of immunotherapy and the molecular mechanism of this process.

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Abbreviations

3MA:

3-Methyladenine

ATG5:

Autophagy protein 5

ATG16L1:

Autophagy-related protein 16-1

CTL:

Cytotoxic T lymphocyte

CTLA4:

Cytotoxic T lymphocyte antigen 4

DAMP:

Damage-associated molecular pattern

DC:

Dendritic cell

GrzB:

Granzyme B

Gy:

Gray

IGFII:

Insulin-like growth factor II

iNOS:

Inducible nitric oxide synthase

LAMP2:

Lysosome-associated membrane protein 2

LLC:

Lewis lung cancer

LC3:

Microtubule-associated protein 1A/1B-light chain 3

MPR:

Mannose-6-phopsphate receptor

PD-L1:

Programmed death-ligand 1

shRNA:

Short hairpin RNA

siRNA:

Small interfering RNA

TAX:

Taxol

TGN:

Trans-golgi network

TLR:

Toll-like receptor

XRT:

Radiation therapy

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Acknowledgments

We thank Dr. Jeffrey Weber for critical reading of the manuscript. This work was supported in part by pilot funds from Donald A. Adam Comprehensive Melanoma Research Center at H. Lee Moffitt Cancer Center and by NIH grant CA168536 to Dmitry I. Gabrilovich. This work was supported in part by microscopy core of H. Lee Moffitt Cancer Center.

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Authors declare no conflicts of interests.

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Authors and Affiliations

  1. H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, 33647, USA

    Sungjune Kim, Rupal Ramakrishnan, Prakash Chinnaiyan, Nikhil Rao, Erin Fowler & John Heine

  2. The Wistar Institute, 3600 Spruce Str., Rm. 118, Philadelphia, PA, 19104, USA

    Sergio Lavilla-Alonso & Dmitry I. Gabrilovich

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  1. Sungjune Kim
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  2. Rupal Ramakrishnan
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  3. Sergio Lavilla-Alonso
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Correspondence to Dmitry I. Gabrilovich.

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Sungjune Kim and Rupal Ramakrishnan have equally contributed to the work.

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Kim, S., Ramakrishnan, R., Lavilla-Alonso, S. et al. Radiation-induced autophagy potentiates immunotherapy of cancer via up-regulation of mannose 6-phosphate receptor on tumor cells in mice. Cancer Immunol Immunother 63, 1009–1021 (2014). https://doi.org/10.1007/s00262-014-1573-4

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