Reciprocal cellular cross-talk within the tumor microenvironment promotes oncolytic virus activity

Abstract

Tumors are complex ecosystems composed of networks of interacting 'normal' and malignant cells. It is well recognized that cytokine-mediated cross-talk between normal stromal cells, including cancer-associated fibroblasts (CAFs), vascular endothelial cells, immune cells, and cancer cells, influences all aspects of tumor biology1. Here we demonstrate that the cross-talk between CAFs and cancer cells leads to enhanced growth of oncolytic virus (OV)-based therapeutics. Transforming growth factor-β (TGF-β) produced by tumor cells reprogrammed CAFs, dampened their steady-state level of antiviral transcripts and rendered them sensitive to virus infection. In turn, CAFs produced high levels of fibroblast growth factor 2 (FGF2), initiating a signaling cascade in cancer cells that reduced retinoic acid-inducible gene I (RIG-I) expression and impeded the ability of malignant cells to detect and respond to virus. In xenografts derived from individuals with pancreatic cancer, the expression of FGF2 correlated with the susceptibility of the cancer cells to OV infection, and local application of FGF2 to resistant tumor samples sensitized them to virotherapy both in vitro and in vivo. An OV engineered to express FGF2 was safe in tumor-bearing mice, showed improved therapeutic efficacy compared to parental virus and merits consideration for clinical testing.

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Figure 1: CAFs, but not NFs, are sensitive to virus-based therapy due to rewired antiviral networks.
Figure 2: Paracrine factors secreted by cancer cells and CAFs enhance OV replication.
Figure 3: FGF2 induces replication of various clinically relevant OVs in a panel of cancer cells and activated fibroblasts.
Figure 4: FGF2 induces sensitivity to OV therapy in vivo.

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References

  1. 1

    Hanahan, D. & Coussens, L.M. Accessories to the crime: functions of cells recruited to the tumor microenvironment. Cancer Cell 21, 309–322 (2012).

    CAS  Article  Google Scholar 

  2. 2

    Chung, A.S. et al. An interleukin-17–mediated paracrine network promotes tumor resistance to anti-angiogenic therapy. Nat. Med. 19, 1114–1123 (2013).

    CAS  Article  Google Scholar 

  3. 3

    Hwang, R.F. et al. Cancer-associated stromal fibroblasts promote pancreatic tumor progression. Cancer Res. 68, 918–926 (2008).

    CAS  Article  Google Scholar 

  4. 4

    Lisanti, M.P., Martinez-Outschoorn, U.E. & Sotgia, F. Oncogenes induce the cancer-associated fibroblast phenotype: Metabolic symbiosis and “fibroblast addiction” are new therapeutic targets for drug discovery. Cell Cycle 12, 2723–2732 (2013).

    CAS  Article  Google Scholar 

  5. 5

    Nakasone, E.S. et al. Imaging tumor-stroma interactions during chemotherapy reveals contributions of the microenvironment to resistance. Cancer Cell 21, 488–503 (2012).

    CAS  Article  Google Scholar 

  6. 6

    Straussman, R. et al. Tumour micro-environment elicits innate resistance to RAF inhibitors through HGF secretion. Nature 487, 500–504 (2012).

    CAS  Article  Google Scholar 

  7. 7

    Sun, Y. et al. Treatment-induced damage to the tumor microenvironment promotes prostate cancer therapy resistance through WNT16B. Nat. Med. 18, 1359–1368 (2012).

    CAS  Article  Google Scholar 

  8. 8

    Wilson, T.R. et al. Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors. Nature 487, 505–509 (2012).

    CAS  Article  Google Scholar 

  9. 9

    Breitbach, C.J. et al. Intravenous delivery of a multi-mechanistic cancer-targeted oncolytic poxvirus in humans. Nature 477, 99–102 (2011).

    CAS  Article  Google Scholar 

  10. 10

    Heo, J. et al. Randomized dose-finding clinical trial of oncolytic immunotherapeutic vaccinia JX-594 in liver cancer. Nat. Med. 19, 329–336 (2013).

    CAS  Article  Google Scholar 

  11. 11

    Kalluri, R. & Zeisberg, M. Fibroblasts in cancer. Nat. Rev. Cancer 6, 392–401 (2006).

    CAS  Article  Google Scholar 

  12. 12

    Löhr, M. et al. Transforming growth factor-β1 induces desmoplasia in an experimental model of human pancreatic carcinoma. Cancer Res. 61, 550–555 (2001).

    PubMed  Google Scholar 

  13. 13

    Rønnov-Jessen, L. & Petersen, O.W. Induction of αα-smooth muscle actin by transforming growth factor-β1 in quiescent human breast gland fibroblasts. Implications for myofibroblast generation in breast neoplasia. Lab. Invest. 68, 696–707 (1993).

    PubMed  Google Scholar 

  14. 14

    Sobol, P.T. et al. Adaptive antiviral immunity is a determinant of the therapeutic success of oncolytic virotherapy. Mol. Ther. 19, 335–344 (2011).

    CAS  Article  Google Scholar 

  15. 15

    Stojdl, D.F. et al. VSV strains with defects in their ability to shutdown innate immunity are potent systemic anti-cancer agents. Cancer Cell 4, 263–275 (2003).

    CAS  Article  Google Scholar 

  16. 16

    Brun, J. et al. Identification of genetically modified Maraba virus as an oncolytic rhabdovirus. Mol. Ther. 18, 1440–1449 (2010).

    CAS  Article  Google Scholar 

  17. 17

    Azzarone, B. et al. Abnormal properties of skin fibroblasts from patients with breast cancer. Int. J. Cancer. 33, 759–764 (1984).

    CAS  Article  Google Scholar 

  18. 18

    Brouty-Boyé, D. et al. Fetal myofibroblast-like cells isolated from post-radiation fibrosis in human breast cancer. Int. J. Cancer. 47, 697–702 (1991).

    Article  Google Scholar 

  19. 19

    Schor, S.L., Schor, A.M. & Rushton, G. Fibroblasts from cancer patients display a mixture of both foetal and adult-like phenotypic characteristics. J. Cell Sci. 90, 401–407 (1988).

    PubMed  Google Scholar 

  20. 20

    Cirri, P. & Chiarugi, P. Cancer associated fibroblasts: the dark side of the coin. Am. J. Cancer Res. 1, 482–497 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  21. 21

    Erez, N., Truitt, M., Olson, P., Arron, S.T. & Hanahan, D. Cancer-associated fibroblasts are activated in incipient neoplasia to orchestrate tumor-promoting inflammation in an NF-κB–dependent manner. Cancer Cell 17, 135–147 (2010).

    CAS  Article  Google Scholar 

  22. 22

    Franco, O.E., Shaw, A.K., Strand, D.W. & Hayward, S.W. Cancer associated fibroblasts in cancer pathogenesis. Semin. Cell Dev. Biol. 21, 33–39 (2010).

    CAS  Article  Google Scholar 

  23. 23

    Orimo, A. et al. Stromal fibroblasts present in invasive human breast carcinomas promote tumor growth and angiogenesis through elevated SDF-1/CXCL12 secretion. Cell 121, 335–348 (2005).

    CAS  Article  Google Scholar 

  24. 24

    Räsänen, K. & Vaheri, A. Activation of fibroblasts in cancer stroma. Exp. Cell Res. 316, 2713–2722 (2010).

    Article  Google Scholar 

  25. 25

    Spaeth, E.L. et al. Mesenchymal stem cell transition to tumor-associated fibroblasts contributes to fibrovascular network expansion and tumor progression. PLoS ONE 4, e4992 (2009).

    Article  Google Scholar 

  26. 26

    Ablasser, A. et al. RIG-I-dependent sensing of poly(dA:dT) through the induction of an RNA polymerase III––transcribed RNA intermediate. Nat. Immunol. 10, 1065–1072 (2009).

    CAS  Article  Google Scholar 

  27. 27

    Chiu, Y.H., Macmillan, J.B. & Chen, Z.J. RNA polymerase III detects cytosolic DNA and induces type I interferons through the RIG-I pathway. Cell 138, 576–591 (2009).

    CAS  Article  Google Scholar 

  28. 28

    Yoneyama, M. et al. The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nat. Immunol. 5, 730–737 (2004).

    CAS  Article  Google Scholar 

  29. 29

    Altomonte, J. & Ebert, O. Sorting out Pandora's box: discerning the dynamic roles of liver microenvironment in oncolytic virus therapy for hepatocellular carcinoma. Front. Oncol. 4, 85 (2014).

    Article  Google Scholar 

  30. 30

    Kurozumi, K. et al. Effect of tumor microenvironment modulation on the efficacy of oncolytic virus therapy. J. Natl. Cancer Inst. 99, 1768–1781 (2007).

    CAS  Article  Google Scholar 

  31. 31

    Stanford, M.M., Breitbach, C.J., Bell, J.C. & McFadden, G. Innate immunity, tumor microenvironment and oncolytic virus therapy: friends or foes? Curr. Opin. Mol. Ther. 10, 32–37 (2008).

    CAS  Google Scholar 

  32. 32

    Kim, R., Emi, M. & Tanabe, K. Cancer immunoediting from immune surveillance to immune escape. Immunology 121, 1–14 (2007).

    CAS  Article  Google Scholar 

  33. 33

    Lichty, B.D., Breitbach, C.J., Stojdl, D.F. & Bell, J.C. Going viral with cancer immunotherapy. Nat. Rev. Cancer 14, 559–567 (2014).

    CAS  Article  Google Scholar 

  34. 34

    Erkan, M. et al. The role of stroma in pancreatic cancer: diagnostic and therapeutic implications. Nat Rev Gastroenterol. Hepatol. 9, 454–467 (2012).

    CAS  Article  Google Scholar 

  35. 35

    Erkan, M. et al. The impact of the activated stroma on pancreatic ductal adenocarcinoma biology and therapy resistance. Curr. Mol. Med. 12, 288–303 (2012).

    CAS  Article  Google Scholar 

  36. 36

    Sanfilippo, C.M. & Blaho, J.A. ICP0 gene expression is a herpes simplex virus type 1 apoptotic trigger. J. Virol. 80, 6810–6821 (2006).

    CAS  Article  Google Scholar 

  37. 37

    Dayer, A.G. et al. Expression of FGF-2 in neural progenitor cells enhances their potential for cellular brain repair in the rodent cortex. Brain 130, 2962–2976 (2007).

    Article  Google Scholar 

  38. 38

    Kim, J.H. et al. Systemic armed oncolytic and immunologic therapy for cancer with JX-594, a targeted poxvirus expressing GM-CSF. Mol. Ther. 14, 361–370 (2006).

    CAS  Article  Google Scholar 

  39. 39

    Breitbach, C.J. et al. Targeting tumor vasculature with an oncolytic virus. Mol. Ther. 19, 886–894 (2011).

    CAS  Article  Google Scholar 

  40. 40

    Emig, D. et al. AltAnalyze and DomainGraph: analyzing and visualizing exon expression data. Nucleic Acids Res. 38, W755–W762 (2010).

    CAS  Article  Google Scholar 

  41. 41

    Eden, E., Navon, R., Steinfeld, I., Lipson, D. & Yakhini, Z. GOrilla: a tool for discovery and visualization of enriched GO terms in ranked gene lists. BMC Bioinformatics 10, 48 (2009).

    Article  Google Scholar 

  42. 42

    Samarajiwa, S.A., Forster, S., Auchettl, K. & Hertzog, P.J. INTERFEROME: the database of interferon regulated genes. Nucleic Acids Res. 37, D852–D857 (2009).

    CAS  Article  Google Scholar 

  43. 43

    Pfaffl, M.W. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29, e45 (2001).

    CAS  Article  Google Scholar 

  44. 44

    Östman, A. & Augsten, M. Cancer-associated fibroblasts and tumor growth–bystanders turning into key players. Curr. Opin. Genet. Dev. 19, 67–73 (2009).

    Article  Google Scholar 

  45. 45

    Tomayko, M.M. & Reynolds, C.P. Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother. Pharmacol. 24, 148–154 (1989).

    CAS  Article  Google Scholar 

  46. 46

    Dell, R.B., Holleran, S. & Ramakrishnan, R. Sample size determination. ILAR J 43, 207–213 (2002).

    CAS  Article  Google Scholar 

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Acknowledgements

This work was funded by grants from the Terry Fox Research Foundation (201201TFF-271514-TFF-AYDP-29782) and the Canadian Institutes of Health Research (314043) to J.C.B., J.-S.D., D.F.S., B.D.L. and R.C.A. C.S.I. is the recipient of a Fellowship award from the Alberta Innovative Health Solutions. J.C.B., D.F.S. and B.D.L. are supported by the Ontario Institute for Cancer Research and the Ottawa Regional Cancer Foundation. M.M. is funded by the Canadian Institutes of Health Research Frederick Banting and Charles Best Master's Award. C.B. is funded by the Natural Sciences and Engineering Research Council of Canada. We thank C. Cemeus and D. Vaillant for their exceptional technical support as well as members of the Bell, Auer, Atkins and Diallo laboratories for feedback on this project. pWPI-spbFGF plasmid was a gift from J. Kiss and P. Salmon (University of Geneva Medical School). HSV-1 N212 expressing GFP14,36 was a gift from K. Mossman (McMaster University). Reovirus was a gift from P. Lee (Dalhousie University). Rabbit anti-reovirus T3 antibody was a gift from E. Brown (University of Ottawa).

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C.S.I., M.M., C.B., D.B.N., S.C., F.L.B., S.H.T., J.M., M. Boileau, D.B., L.S., P.S., H.L.A. and V.A.J. conducted in vitro experiments. C.S.I., M.M., T.F., C.T.d.S. and D.B.N. performed mouse experiments. P.P. and A.C. recruited individuals with pancreatic cancer and collected biopsies from subjects with pancreatic cancer. R.M.S., M. Burdick and A.C. isolated normal and cancer-associated fibroblasts. A.A., J.Z. and R.C.A. engineered the MG1-FGF2 virus. C.S.I., M.M., C.B., A.A., C.L.A., D.F.S., J.-S.D., B.D.L. and J.C.B. designed the study and were involved in writing of the manuscript.

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Correspondence to John C Bell.

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Ilkow, C., Marguerie, M., Batenchuk, C. et al. Reciprocal cellular cross-talk within the tumor microenvironment promotes oncolytic virus activity. Nat Med 21, 530–536 (2015). https://doi.org/10.1038/nm.3848

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