Preclinical evaluation of radiation and systemic, RGD-targeted, adeno-associated virus phage-TNF gene therapy in a mouse model of spontaneously metastatic melanoma

Article metrics


The incidence of melanoma in the United States continues to rise, with metastatic lesions notoriously recalcitrant to therapy. There are limited effective treatment options available and a great need for more effective therapies that can be rapidly integrated in the clinic. In this study, we demonstrate that the combination of RGD-targeted adeno-associated virus phage (RGD-AAVP-TNF) with hypofractionated radiation therapy results in synergistic inhibition of primary syngeneic B16 melanoma in a C57 mouse model. Furthermore, this combination appeared to modify the tumor microenvironment, resulting in decreased Tregs in the draining LN and increased tumor-associated macrophages within the primary tumor. Finally, there appeared to be a reduction in metastatic potential and a prolongation of overall survival in the combined treatment group. These results indicate the use of targeted TNF gene therapy vector with radiation treatment could be a valuable treatment option for patients with metastatic melanoma.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5


  1. 1

    Howlader N, Noone A, Krapcho M, Garshell J, Neyman N, Altekruse S et al. SEER cancer statistics review. Surveill Epidemiol End Results Program 1975-2010, NCI, Bethesda, MD, November 2012.

  2. 2

    Markovic SN, Erickson LA, Rao RD, Weenig RH, Pockaj BA, Bardia A et al. Malignant melanoma in the 21st century, part 2: staging, prognosis, and treatment. Mayo Clin Proc 2007; 82: 490–513.

  3. 3

    Fife KM, Colman MH, Stevens GN, Firth IC, Moon D, Shannon KF et al. Determinants of outcome in melanoma patients with cerebral metastases. J Clin Oncol 2004; 22: 1293–1300.

  4. 4

    Okwan-Duodu D, Pollack BP, Lawson D, Khan MK . Role of radiation therapy as immune activator in the era of modern immunotherapy for metastatic malignant melanoma. Am J Clin Oncol 2014; 00: 1–7.

  5. 5

    Datta R, Rubin E, Sukhatme V, Qureshi S, Hallahan D, Weichselbaum RR et al. Ionizing radiation activates transcription of the EGR1 gene via CArG elements. Proc Natl Acad Sci USA 1992; 89: 10149–10153.

  6. 6

    Hallahan DE, Mauceri HJ, Seung LP, Dunphy EJ, Wayne JD, Hanna NN et al. Spatial and temporal control of gene therapy using ionizing radiation. Nat Med 1995; 1: 786–791.

  7. 7

    Staba MJ, Mauceri HJ, Kufe DW, Hallahan DE, Weichselbaum RR . Adenoviral TNF-alpha gene therapy and radiation damage tumor vasculature in a human malignant glioma xenograft. Gene Ther 1998; 5: 293–300.

  8. 8

    Chang KJ, Reid T, Senzer N, Swisher S, Pinto H, Hanna N et al. Phase i evaluation of TNFerade biologic plus chemoradiotherapy before esophagectomy for locally advanced resectable esophageal cancer. Gastrointest Endosc 2012; 75: 1139–1146.

  9. 9

    Senzer N, Mani S, Rosemurgy A, Nemunaitis J, Cunningham C, Guha C et al. TNFerade biologic, an adenovector with a radiation-inducible promoter, carrying the human tumor necrosis factor alpha gene: a phase I study in patients with solid tumors. J Clin Oncol 2004; 22: 592–601.

  10. 10

    Moreno-Ramirez D, de la Cruz-Merino L, Ferrandiz L, Villegas-Portero R, Nieto-Garcia A . Isolated limb perfusion for malignant melanoma: systematic review on effectiveness and safety. Oncologist 2010; 15: 416–427.

  11. 11

    Hallahan DE, Beckett MA, Kufe D, Weichselbaum RR . The interaction between recombinant human tumor necrosis factor and radiation in 13 human tumor cell lines. Int J Radiat Oncol Biol Phys 1990; 19: 69–74.

  12. 12

    Yao VJ, Ozawa MG, Varner AS, Kasman IM, Chanthery YH, Pasqualini R et al. Antiangiogenic therapy decreases integrin expression in normalized tumor blood vessels. Cancer Res 2006; 66: 2639–2649.

  13. 13

    Tandle A, Hanna E, Lorang D, Hajitou A, Moya CA, Pasqualini R et al. Tumor vasculature-targeted delivery of tumor necrosis factor-$α$. Cancer 2009; 115: 128–139.

  14. 14

    Yuan Z, Syrkin G, Adem A, Geha R, Pastoriza J, Vrikshajanani C et al. Blockade of inhibitors of apoptosis (IAPs) in combination with tumor-targeted delivery of tumor necrosis factor-alpha leads to synergistic antitumor activity. Cancer Gene Ther 2013; 20: 46–56.

  15. 15

    Teicher BA . Tumor models for efficacy determination. Mol Cancer Ther 2006; 5: 2435–2443.

  16. 16

    Tomayko MM, Reynolds CP . Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother Pharmacol 1989; 24: 148–154.

  17. 17

    Overwijk WW, Restifo NP . B16 as a mouse model for human melanoma. Curr Protoc Immunol 2001 May; Chapter 20:Unit 20.1:1–33.

  18. 18

    Bliss CI . The toxicity of poisons applied jointly. Ann Appl Biol 1939; 26: 585–615.

  19. 19

    Hou JY, Rodriguez-Gabin A, Samaweera L, Hazan R, Goldberg GL, Horwitz SB et al. Exploiting MEK inhibitor-mediated activation of ER?? for therapeutic intervention in ER-positive ovarian carcinoma. PLoS One 2013; 8(2): e54103 (1-10).

  20. 20

    Dunn GP, Bruce AT, Ikeda H, Old LJ, Schreiber RD . Cancer immunoediting: from immunosurveillance to tumor escape. Nat Immunol 2002; 3: 991–998.

  21. 21

    Dunn GP, Old LJ, Schreiber RD . The immunobiology of cancer immunosurveillance and immunoediting. Immunity 2004; 21: 137–148.

Download references

Author information

Correspondence to Z Yuan or C Guha.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark