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Hypoxia-induced autophagy of stellate cells inhibits expression and secretion of lumican into microenvironment of pancreatic ductal adenocarcinoma

Cell Death & Differentiation (2018) | Download Citation

Abstract

Lumican is secreted by pancreatic stellate cells and inhibits cancer progression. Extracellular lumican inhibits cancer cell replication and restrains growth of early-stage pancreatic adenocarcinoma (PDAC) such that patients with tumors containing stromal lumican experience a three-fold longer survival after treatment. In the present study, patient tumor tissues, ex-vivo cultures of patient-derived xenografts (PDX), PDAC stellate and tumor cells were used to investigate whether hypoxia (1% O2) within the tumor microenvironment influences stromal lumican expression and secretion. We observed that hypoxia significantly reduced lumican expression and secretion from pancreatic stellate cells, but not cancer cells. Although hypoxia enhanced lactate dehydrogenase A (LDHA) expression and lactate secretion from all cells, neither hypoxia-induced nor exogenous lactate influenced lumican expression. Autophagy was induced by hypoxia in ex vivo cultures of PDX and pancreatic stellate cells, but not cancer cells cultured in 2D. Autophagic flux inhibitors, bafilomycin A1, chloroquine diphosphate salt, and ammonium chloride prevented hypoxia-mediated reduction in lumican expression in stellate cells. Furthermore, inhibition of AMP-regulated protein kinase (AMPK) phosphorylation or hypoxia-inducible factor (HIF)-1α expression within hypoxic stellate cells restored lumican expression levels. Hypoxia did not affect lumican mRNA expression, indicating that hypoxia-induced reduction of lumican occurs post-transcriptionally; in addition, AMPK inhibition prevented hypoxia-reduced phosphorylation of the mTOR/p70S6K/4EBP signaling pathway, a key contributor to protein synthesis. Taken together, these findings demonstrate that hypoxia reduces stromal lumican in PDAC through autophagy-mediated degradation and reduction in protein synthesis within pancreatic cancer stellate cells.

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Acknowledgements

This work was supported by grants from the Skip Viragh Family Foundation (to JBF), the W. Smith Foundation (to JBF), National Institutes of Health (NIH) grant T32CA009599 (to MRP), an in-kind grant from the Center for Advanced Biomedical Imaging and GE Healthcare (to EJK and JBF), and U54 CA210181-01 “Center for Immunotherapeutic Transport Oncophysics (CITO)” grant (to Koay, Fleming and Brekken).

Funding

This work was supported by grants from the Skip Viragh Family Foundation (to JBF), the W. Smith Foundation (to JBF), National Institutes of Health (NIH) grant T32CA009599 (to MRP), an in-kind grant from the Center for Advanced Biomedical Imaging and GE Healthcare (to EJK and JBF), and U54 CA210181-01 “Center for Immunotherapeutic Transport Oncophysics (CITO)” grant (to Koay, Fleming and Brekken).

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Affiliations

  1. Department of Surgical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    • Xinqun Li
    • , Ya’an Kang
    • , Bingbing Dai
    • , Mayrim Rios Perez
    • , Michael Pratt
    • , Michael Kim
    •  & Jason B. Fleming
  2. Division of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    • Yeonju Lee
    •  & Eugene J. Koay
  3. Hamon Center of Therapeutic Oncology Research and Division of Surgical Oncology, Department of Surgery, University of Texas Southwestern Medical Center, Dallas, TX, USA

    • Rolf A. Brekken
  4. Department of Gastrointestinal Oncology, Moffitt Cancer Center, Tampa, FL, USA

    • Jason B. Fleming

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Conflict of interest

The authors declare that they have no conflict of interest.

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Correspondence to Jason B. Fleming.

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DOI

https://doi.org/10.1038/s41418-018-0207-3

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Edited by G. Del Sal