Cancer Immunology, Immunotherapy

, Volume 65, Issue 5, pp 613–624 | Cite as

Anti-tumor effects of DNA vaccine targeting human fibroblast activation protein α by producing specific immune responses and altering tumor microenvironment in the 4T1 murine breast cancer model

  • Qiu Xia
  • Fang-Fang Zhang
  • Fei Geng
  • Chen-Lu Liu
  • Ping Xu
  • Zhen-Zhen Lu
  • Bin Yu
  • Hui Wu
  • Jia-Xin Wu
  • Hai-Hong Zhang
  • Wei Kong
  • Xiang-Hui Yu
Original Article
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Abstract

Fibroblast activation protein α (FAPα) is a tumor stromal antigen overexpressed by cancer-associated fibroblasts (CAFs). CAFs are genetically more stable compared with the tumor cells and immunosuppressive components of the tumor microenvironment, rendering them excellent targets for cancer immunotherapy. DNA vaccines are widely applied due to their safety. To specifically destroy CAFs, we constructed and examined the immunogenicity and anti-tumor immune mechanism of a DNA vaccine expressing human FAPα. This vaccine successfully reduced 4T1 tumor growth through producing FAPα-specific cytotoxic T lymphocyte responses which could kill CAFs, and the decrease in FAPα-expressing CAFs resulted in markedly attenuated expression of collagen I and other stromal factors that benefit the tumor progression. Based on these results, a DNA vaccine targeting human FAPα may be an attractive and effective cancer immunotherapy strategy.

Keywords

FAPα CAF Immune suppression DNA vaccine Immunotherapy 

Abbreviations

CAFs

Cancer-associated fibroblasts

ECM

Extracellular matrix

FAPα

Fibroblast activation protein α

FGF-2

Fibroblast growth factor-2

HGF

Hematopoietic growth factor

PDGF

Platelet-derived growth factor

SDF-1

Stromal cell-derived factor-1

VEGFα

Vascular endothelial growth factor alpha

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Notes

Acknowledgments

This study was supported by the National Natural Science Foundation of China (No. 31300765), Doctoral Program of Higher Education (New Teachers) (No. 20120061120025), Jilin Province Science and Technology Development Program (no. 20130522006JH), the National Science and Technology Major Project of the Ministry of Science and Technology of China (Nos. 2014ZX09304314-001, 2012ZX10001009-12) and the Fundamental Research Funds for the Central Universities (No. JCKY-QKJC03).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Siegel R, Ma J, Zou Z, Jemal A (2014) Cancer statistics, 2014. CA Cancer J Clin 64:9–29CrossRefPubMedGoogle Scholar
  2. 2.
    Prud’homme GJ (2005) DNA vaccination against tumors. J Gene Med 7:3–17CrossRefPubMedGoogle Scholar
  3. 3.
    Nguyen T, Urban J, Kalinski P (2014) Therapeutic cancer vaccines and combination immunotherapies involving vaccination. Immunotargets Ther 3:135–150Google Scholar
  4. 4.
    Scanlan MJ, Gure AO, Jungbluth AA, Old LJ, Chen Y-T (2002) Cancer/testis antigens: an expanding family of targets for cancer immunotherapy. Immunol Rev 188:22–32CrossRefPubMedGoogle Scholar
  5. 5.
    Acharya PS, Zukas A, Chandan V, Katzenstein A-LA, Puré E (2006) Fibroblast activation protein: a serine protease expressed at the remodeling interface in idiopathic pulmonary fibrosis. Hum Pathol 37:352–360CrossRefPubMedGoogle Scholar
  6. 6.
    Garin-Chesa P, Old LJ, Rettig WJ (1990) Cell surface glycoprotein of reactive stromal fibroblasts as a potential antibody target in human epithelial cancers. Proc Natl Acad Sci USA 87:7235–7239CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Niedermeyer J, Scanlan MJ, Garin-Chesa P, Daiber C, Fiebig HH, Old LJ, Rettig WJ, Schnapp A (1997) Mouse fibroblast activation protein: molecular cloning, alternative splicing and expression in the reactive stroma of epithelial cancers. Int J Cancer 71:383–389CrossRefPubMedGoogle Scholar
  8. 8.
    Goldstein LA, Ghersi G, Piñeiro-Sánchez ML, Salamone M, Yeh Y, Flessate D, Chen W-T (1997) Molecular cloning of seprase: a serine integral membrane protease from human melanoma. Biochim Biophys Acta 1361:11–19CrossRefPubMedGoogle Scholar
  9. 9.
    Rettig WJ, Garin-Chesa P, Healey JH, Su SL, Ozer HL, Schwab M, Albino AP, Old LJ (1993) Regulation and heteromeric structure of the fibroblast activation protein in normal and transformed cells of mesenchymal and neuroectodermal origin. Cancer Res 53:3327–3335PubMedGoogle Scholar
  10. 10.
    Micke P (2004) Tumour-stroma interaction: Cancer-associated fibroblasts as novel targets in anti-cancer therapy? Lung Cancer 45:S163–S175CrossRefPubMedGoogle Scholar
  11. 11.
    Östman A, Augsten M (2009) Cancer-associated fibroblasts and tumor growth—bystanders turning into key players. Curr Opin Genet Dev 19:67–73CrossRefPubMedGoogle Scholar
  12. 12.
    Lee J, Fassnacht M, Nair S, Boczkowski D, Gilboa E (2005) Tumor immunotherapy targeting fibroblast activation protein, a product expressed in tumor-associated fibroblasts. Cancer Res 65:11156–11163CrossRefPubMedGoogle Scholar
  13. 13.
    Cheng JD, Dunbrack RL, Valianou M, Rogatko A, Alpaugh RK, Weiner LM (2002) Promotion of tumor growth by murine fibroblast activation protein, a serine protease, in an animal model. Cancer Res 62:4767–4772PubMedGoogle Scholar
  14. 14.
    Wen Y, Wang CT, Ma TT et al (2010) Immunotherapy targeting fibroblast activation protein inhibits tumor growth and increases survival in a murine colon cancer model. Cancer Sci 101:2325–2332CrossRefPubMedGoogle Scholar
  15. 15.
    Fassnacht M, Lee J, Milazzo C, Boczkowski D, Su Z, Nair S, Gilboa E (2005) Induction of CD4+ and CD8+ T-cell responses to the human stromal antigen, fibroblast activation protein: implication for cancer immunotherapy. Clin Cancer Res 11:5566–5571CrossRefPubMedGoogle Scholar
  16. 16.
    Fearon DT (2014) The carcinoma-associated fibroblast expressing fibroblast activation protein and escape from immune surveillance. Cancer Immunol Res 2:187–193CrossRefPubMedGoogle Scholar
  17. 17.
    Kalluri R, Zeisberg M (2006) Fibroblasts in cancer. Nat Rev Cancer 6:392–401CrossRefPubMedGoogle Scholar
  18. 18.
    Ribatti D, Vacca A (2008) The role of microenvironment in tumor angiogenesis. Genes Nutr 3:29–34CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Joyce JA, Fearon DT (2015) T cell exclusion, immune privilege, and the tumor microenvironment. Science 348:74–80CrossRefPubMedGoogle Scholar
  20. 20.
    Marincola FM, Jaffee EM, Hicklin DJ, Ferrone S (2000) Escape of human solid tumors from T-cell recognition: molecular mechanisms and functional significance. Adv Immunol 74:181–273CrossRefPubMedGoogle Scholar
  21. 21.
    O’Connor DS, Schechner JS, Adida C, Mesri M, Rothermel AL, Li F, Nath AK, Pober JS, Altieri DC (2000) Control of apoptosis during angiogenesis by survivin expression in endothelial cells. Am J Pathol 156:393–398CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Gabrilovich DI, Chen HL, Girgis KR, Cunningham HT, Meny GM, Nadaf S, Kavanaugh D, Carbone DP (1996) Production of vascular endothelial growth factor by human tumors inhibits the functional maturation of dendritic cells. Nat Med 2:1096–1103CrossRefPubMedGoogle Scholar
  23. 23.
    Elgert KD, Alleva DG, Mullins DW (1998) Tumor-induced immune dysfunction: the macrophage connection. J Leukoc Biol 64:275–290PubMedGoogle Scholar
  24. 24.
    Kraman M, Bambrough PJ, Arnold JN, Roberts EW, Magiera L, Jones JO, Gopinathan A, Tuveson DA, Fearon DT (2010) Suppression of antitumor immunity by stromal cells expressing fibroblast activation protein-α. Science 330:827–830CrossRefPubMedGoogle Scholar
  25. 25.
    LeBleu VS (2015) Imaging the tumor microenvironment. Cancer J 21:174–178CrossRefPubMedGoogle Scholar
  26. 26.
    Liu R, Li H, Liu L, Yu J, Ren X (2012) Fibroblast activation protein: a potential therapeutic target in cancer. Cancer Biol Ther 13:123–129CrossRefPubMedGoogle Scholar
  27. 27.
    Cheng JD, Valianou M, Canutescu AA, Jaffe EK, Lee H-O, Wang H, Lai JH, Bachovchin WW, Weiner LM (2005) Abrogation of fibroblast activation protein enzymatic activity attenuates tumor growth. Mol Cancer Ther 4:351–360CrossRefPubMedGoogle Scholar
  28. 28.
    Lladser A, Ljungberg K, Tufvesson H, Tazzari M, Roos A-K, Quest AF, Kiessling R (2010) Intradermal DNA electroporation induces survivin-specific CTLs, suppresses angiogenesis and confers protection against mouse melanoma. Cancer Immunol Immunother 59:81–92CrossRefPubMedGoogle Scholar
  29. 29.
    Nagaraj S, Pisarev V, Kinarsky L, Sherman S, Muro-Cacho C, Altieri DC, Gabrilovich DI (2007) Dendritic cell-based full-length survivin vaccine in treatment of experimental tumors. J Immunother 30:169–179CrossRefPubMedGoogle Scholar
  30. 30.
    Wang Y-Q, Zhang H-H, Liu C-L et al (2013) Enhancement of survivin-specific anti-tumor immunity by adenovirus prime protein-boost immunity strategy with DDA/MPL adjuvant in a murine melanoma model. Int Immunopharmacol 17:9–17CrossRefPubMedGoogle Scholar
  31. 31.
    Wang Y, Liu C, Xia Q et al (2014) Antitumor effect of adenoviral vector prime protein boost immunity targeting the MUC1 VNTRs. Oncol Rep 31:1437–1444PubMedGoogle Scholar
  32. 32.
    Zhang H, Wang Y, Liu C et al (2012) DNA and adenovirus tumor vaccine expressing truncated survivin generates specific immune responses and anti-tumor effects in a murine melanoma model. Cancer Immunol Immunother 61:1857–1867CrossRefPubMedGoogle Scholar
  33. 33.
    You Q, Jiang C, Wu Y et al (2012) Subcutaneous administration of modified vaccinia virus ankara expressing an Ag85B-ESAT6 fusion protein, but not an adenovirus-based vaccine, protects mice against intravenous challenge with Mycobacterium tuberculosis. Scand J Immunol 75:77–84CrossRefPubMedGoogle Scholar
  34. 34.
    Loeffler M, Kruger JA, Niethammer AG, Reisfeld RA (2006) Targeting tumor-associated fibroblasts improves cancer chemotherapy by increasing intratumoral drug uptake. J Clin Invest 116:1955–1962. doi: 10.1172/JCI26532 CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Liao D, Luo Y, Markowitz D, Xiang R, Reisfeld RA (2009) Cancer associated fibroblasts promote tumor growth and metastasis by modulating the tumor immune microenvironment in a 4T1 murine breast cancer model. PLoS One 4:e7965CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Henry LR, Lee H-O, Lee JS, Klein-Szanto A, Watts P, Ross EA, Chen W-T, Cheng JD (2007) Clinical implications of fibroblast activation protein in patients with colon cancer. Clin Cancer Res 13:1736–1741CrossRefPubMedGoogle Scholar
  37. 37.
    Hofheinz R-D, Al-Batran S-E, Hartmann F et al (2003) Stromal antigen targeting by a humanised monoclonal antibody: an early phase II trial of sibrotuzumab in patients with metastatic colorectal cancer. Onkologie 26:44–48CrossRefPubMedGoogle Scholar
  38. 38.
    Scott AM, Wiseman G, Welt S et al (2003) A phase I dose-escalation study of sibrotuzumab in patients with advanced or metastatic fibroblast activation protein-positive cancer. Clin Cancer Res 9:1639–1647PubMedGoogle Scholar
  39. 39.
    Niedermeyer J, Kriz M, Hilberg F et al (2000) Targeted disruption of mouse fibroblast activation protein. Mol Cell Biol 20:1089–1094CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Park JE, Lenter MC, Zimmermann RN, Garin-Chesa P, Old LJ, Rettig WJ (1999) Fibroblast activation protein, a dual specificity serine protease expressed in reactive human tumor stromal fibroblasts. J Biol Chem 274:36505–36512CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Qiu Xia
    • 1
  • Fang-Fang Zhang
    • 1
  • Fei Geng
    • 1
  • Chen-Lu Liu
    • 1
  • Ping Xu
    • 1
  • Zhen-Zhen Lu
    • 1
  • Bin Yu
    • 1
  • Hui Wu
    • 1
  • Jia-Xin Wu
    • 1
  • Hai-Hong Zhang
    • 1
  • Wei Kong
    • 1
  • Xiang-Hui Yu
    • 1
  1. 1.National Engineering Laboratory for AIDS Vaccine, School of Life ScienceJilin UniversityChangchunPeople’s Republic of China

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