Analytical and Bioanalytical Chemistry

, Volume 411, Issue 4, pp 925–933 | Cite as

Engineering oncolytic vaccinia virus with functional peptides through mild and universal strategy

  • Li-Li Huang
  • Xue Li
  • Kejiang Liu
  • Binsuo Zou
  • Hai-Yan XieEmail author
Research Paper


Oncolytic virotherapy is one of promising tumor therapy modalities. However, its therapeutic efficacy is still limited due to the immunogenicity and poor tumor-targeting capability. In this report, an engineered oncolytic vaccinia virus (OVV) was constructed by site-specifically introducing azide groups to the envelope of OVV during the in situ assembling process of virions. Subsequently, dibenzocyclooctynes (DBCO) derivate T7 peptide and DBCO derivate self-peptide were simultaneously conjugated to the azide-modified OVV (azide-OVV) via copper-free click chemistry. The infectivity of peptide-conjugated virus was well kept. Meanwhile, both of the targeting capacity to transferrin receptor (TfR)-overexpressed tumor cells and the in vivo blood circulation time increased. Therefore, the growth of TfR-positive tumor could be significantly inhibited after intravenously injecting the engineered OVV, while no noticeable side effects. This construction strategy can be popularized to other enveloped oncolytic virus (OV), thus a universal engineering platform can be provided for OV cancer therapy.

Graphical Abstract

An engineered oncolytic vaccinia virus (OVV) was constructed by bioconjugating DBCO derivate T7 peptide and DBCO derivate self-peptide with azide-modified OVV via copper-free click chemistry. As a result, the tumor inhibit effect was significantly enhanced attributed to the prolonged in vivo circulation time and improved targeting recognition capability.


Oncolytic virotherapy T7 peptide Self peptide Copper-free click chemistry Azide-enabled oncolytic vaccinia virus Immunogenicity 



We thank Xin-Yuan Liu for providing oncolytic vaccinia virus (OncoPox-IL24-MnSOD). We thank Wei Wei of Institute of Process Engineering, Chinese Academy of Sciences, for his help in animal experiments.

Funding information

This work was supported by the National Science and Technology Major Project (No. 2018ZX 10301405-001), China Postdoctoral Science Foundation (No. 2018M630076), and National Natural Science Foundation of China (No. 21874011).

Compliance with ethical standards

Competing interests

The authors declare that they have no competing interest.


  1. 1.
    Fountzilas C, Patel S, Mahalingam D. Oncolytic virotherapy updates and future directions. Oncotarget. 2017;8(60):102617–39.CrossRefGoogle Scholar
  2. 2.
    Heo J, Reid T, Ruo L, Breitbach CJ, Rose S, Bloomston M, et al. Randomized dose-finding clinical trial of oncolytic immunotherapeutic vaccinia JX-594 in liver cancer. Nat Med. 2013;19(3):329–36.CrossRefGoogle Scholar
  3. 3.
    Ungerechts G, Bossow S, Leuchs B, Holm PS, Rommelaere J, Coffey M, et al. Moving oncolytic viruses into the clinic: clinical-grade production, purification, and characterization of diverse oncolytic viruses. Mol Ther Methods Clin Dev. 2016;3:16018.CrossRefGoogle Scholar
  4. 4.
    Miest TS, Cattaneo R. New viruses for cancer therapy: meeting clinical needs. Nat Rev Microbiol. 2014;12(1):23–34.CrossRefGoogle Scholar
  5. 5.
    Zonov E, Kochneva G, Yunusova A, Grazhdantseva A, Richter V, Ryabchikova E. Features of the antitumor effect of vaccinia virus Lister strain. Viruses. 2016;8(1):1–16.CrossRefGoogle Scholar
  6. 6.
    Wang Y, Yuan M, Ahmed J, Lemoine NR. Oncolytic vaccinia virus. U.S. Patent Application No 15/301, 304, 2015.Google Scholar
  7. 7.
    Delwar ZM, Kuo Y, Wen YH, Rennie PS, Jia W. Oncolytic virotherapy blockade by microglia and macrophages requires STAT1/3. Cancer Res. 2017;78(3):718–30.CrossRefGoogle Scholar
  8. 8.
    Marelli G, Howells A, Lemoine NR, Wang Y. Oncolytic viral therapy and the immune system: a double-edged sword against cancer. Front Immunol. 2018;9(866):1–8.Google Scholar
  9. 9.
    Hung CF, Tsai YC, He L, Coukos G, Fodor I, Qin L, et al. Vaccinia virus preferentially infects and controls human and murine ovarian tumors in mice. Gene Ther. 2007;14(1):20–9.CrossRefGoogle Scholar
  10. 10.
    Cattaneo R, Miest T, Shashkova EV, Barry MA. Reprogrammed viruses as cancer therapeutics: targeted, Armed and Shielded. Nat Rev Microbiol. 2008;6(7):529–40.CrossRefGoogle Scholar
  11. 11.
    Parker AL, Newman C, Briggs S, Seymour L, Sheridan PJ. Nonviral gene delivery: techniques and implications for molecular medicine. Expert Rev Mol Med. 2003;5(22):1–15.CrossRefGoogle Scholar
  12. 12.
    Lee CH, Kasala D, Na Y, Lee MS, Kim SW, Leong JH, et al. Enhanced therapeutic efficacy of an adenovirus-PEI-bile-acid complex in tumors with low coxsackie and adenovirus receptor expression. Biomaterials. 2014;35(21):5505–16.CrossRefGoogle Scholar
  13. 13.
    O'Riordan CR, Lachapelle A, Delgado C, Parkes V, Wadsworth SC, Smith AE, et al. PEGylation of adenovirus with retention of infectivity and protection from neutralizing antibody in vitro and in vivo. Hum Gene Ther. 1999;10(8):1349–58.CrossRefGoogle Scholar
  14. 14.
    Tesfay MZ, Kirk AC, Hadac EM, Griesmann GE, Federspiel MJ, Barber GN, et al. PEGylation of vesicular stomatitis virus extends virus persistence in blood circulation of passively immunized mice. J Virol. 2013;87(7):3752–9.CrossRefGoogle Scholar
  15. 15.
    Park TG, Jeong JH, Kim SW. Current status of polymeric gene delivery systems. Adv Drug Deliv Rev. 2006;58:467–86.CrossRefGoogle Scholar
  16. 16.
    Li S, Huang L. Nonviral gene therapy: promises and challenges. Gene Ther. 2000;7(1):31–4.CrossRefGoogle Scholar
  17. 17.
    Hao J, Huang LL, Zhang R, Wang HZ, Xie HYA. Mild and reliable method to label enveloped virus with quantum dots by copper-free click chemistry. Anal Chem. 2012;84(19):8364–70.CrossRefGoogle Scholar
  18. 18.
    Huang LL, Lu GH, Hao J, Wang H, Yin DL, Xie HY. Enveloped virus labeling via both intrinsic biosynthesis and metabolic incorporation of phospholipids in host cells. Anal Chem. 2013;85(10):5263–70.CrossRefGoogle Scholar
  19. 19.
    Huang LL, Wu LL, Li X, Liu K, Zhao D, Xie HY. Labeling and single particle tracking based entry mechanism study of vaccinia virus from the Tiantan strain. Anal Chem. 2018;90(5):3452–9.CrossRefGoogle Scholar
  20. 20.
    Banerjee PS, Ostapachuk P, Hearing P, Carrico I. Chemoselective attachment of small molecule effector functionality to human adenoviruses facilitates gene delivery to cancer cells. J Am Chem Soc. 2010;132(39):13615–7.CrossRefGoogle Scholar
  21. 21.
    Dissanayake S, Denny WA, Gamage S, Sarojini V. Recent developments in anticancer drug delivery using cell penetrating and tumor targeting peptides. J Control Release. 2017;250:62–76.CrossRefGoogle Scholar
  22. 22.
    Kuang Y, An S, Guo Y, Huang S, Shao K, Liu Y, et al. T7 Peptide-functionalized nanoparticles utilizing rna interference for glioma dual targeting. Int J Pharm. 2013;454:11–20.CrossRefGoogle Scholar
  23. 23.
    Han L, Huang R, Li J, Liu S, Huang S, Jiang C. Plasmid pORF-hTRAIL and doxorubicin co-delivery targeting to tumor using peptide-conjugated polyamidoamine dendrimer. Biomaterials. 2011;32(4):1242–52.CrossRefGoogle Scholar
  24. 24.
    Rodriguez PL, Harada T, Christian DA, Pantano DA, Tsai RK, Discher DE. Minimal “self” peptides that inhibit phagocytic clearance and enhance delivery of nanoparticles. Science. 2013;339(6122):971–5.CrossRefGoogle Scholar
  25. 25.
    Huang LL, Zhou P, Wang HZ, Zhang R, Hao J, Xie HY, et al. A new stable and reliable method for labeling nucleic acids of fully replicative viruses. Chem Commun. 2012;48(18):2424–6.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Li-Li Huang
    • 1
  • Xue Li
    • 1
  • Kejiang Liu
    • 1
  • Binsuo Zou
    • 1
  • Hai-Yan Xie
    • 1
    Email author
  1. 1.School of Life ScienceBeijing Institute of TechnologyBeijingChina

Personalised recommendations