Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Novel strategies to augment genetically delivered immunotoxin molecular therapy for cancer therapy

Cancer Gene Therapy volume 16, pages 861–872 (2009)Cite this article

Abstract

Immunotoxin therapy is a promising molecular cancer treatment strategy. Its main advantage is seletive cytotoxicity towards tumor cells and minimal toxicity in normal tissues. However, a short half-life and rapid clearance severely hampers its clinical application. We report here a novel genetic approach in which a recombinant adenovirus vector was used to deliver an immunotoxin gene e23(scFv)-PE40 targeted to the oncogene c-erbB-2 (also known as Her2/neu). This vector, when combined with a low dose of a conditionally replicative adenovirus vector (CRAd), has enhanced tumor-killing ability either alone or in combination with the chemotherapeutic agent etoposide. Our data show that low-dose CRAd facilitated the replication of replication-deficient Ad-e23(scFv)-PE40 up to 6–20 times and the transcription of e23(scFv)-PE40 gene up to 12 times. Moreover, etoposide increased the e23(scFv)-PE40 transcription up to 8.5 times. Furthermore, we show that systemic application of Ad-e23(scFv)-PE40 and enhanced expression of the immunotoxin gene was well tolerated as determined by serum biochemical markers and histological examination of most vital organs. Taken together, our data support a novel genetic immunotoxin delivery approach that may yield enhanced efficacy against a variety of Her2/neu-expressing tumors.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

Abbreviations

Ad:

adenovirus

CRAd:

conditionally replicative competent adenovirus

FBS:

fetal bovine serum

MOI:

multiplicity of infection

PE:

Pseudomonas exotoxin

scFv:

single-chain fragment variable

TUNEL:

terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling.

References

  1. Burstein HJ . The distinctive nature of HER2-positive breast cancers. N Engl J Med 2005; 353: 1652–1654.

    Article  CAS  PubMed  Google Scholar 

  2. Pritchard KI, Shepherd LE, O’Malley FP, Andrulis IL, Tu D, Bramwell VH et al. HER2 and responsiveness of breast cancer to adjuvant chemotherapy. N Engl J Med 2006; 354: 2103–2111.

    Article  CAS  PubMed  Google Scholar 

  3. Ross JS, Fletcher JA, Linette GP, Stec J, Clark E, Ayers M et al. The Her-2/neu gene and protein in breast cancer 2003: biomarker and target of therapy. Oncologist 2003; 8: 307–325.

    Article  CAS  PubMed  Google Scholar 

  4. Stephens P, Hunter C, Bignell G, Edkins S, Davies H, Teague J et al. Lung cancer: intragenic ERBB2 kinase mutations in tumours. Nature 2004; 431: 525–526.

    Article  CAS  PubMed  Google Scholar 

  5. Swanton C, Futreal A, Eisen T . Her2-targeted therapies in non-small cell lung cancer. Clin Cancer Res 2006; 12: 4377s–4383s.

    Article  CAS  PubMed  Google Scholar 

  6. Slamon DJ, Godolphin W, Jones LA, Holt JA, Wong SG, Keith DE et al. Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 1989; 244: 707–712.

    Article  CAS  PubMed  Google Scholar 

  7. Lui VW, He Y, Goyal K, Huang L . Specific down-regulation of HER-2/neu mediated by a chimeric U6 hammerhead ribozyme results in growth inhibition of human ovarian carcinoma. Mol Ther 2001; 3: 169–177.

    Article  CAS  PubMed  Google Scholar 

  8. Kim MA, Jung EJ, Lee HS, Lee HE, Jeon YK, Yang HK et al. Evaluation of HER-2 gene status in gastric carcinoma using immunohistochemistry, fluorescence in situ hybridization, and real-time quantitative polymerase chain reaction. Hum Pathol 2007; 38: 1386–1393.

    Article  CAS  PubMed  Google Scholar 

  9. Shiau AL, Lin YP, Shieh GS, Su CH, Wu WL, Tsai YS et al. Development of a conditionally replicating Pseudorabies virus for HER-2/neu-overexpressing bladder cancer therapy. Mol Ther 2007; 15: 131–138.

    Article  CAS  PubMed  Google Scholar 

  10. Schlieman MG, Fahy BN, Ramsamooj R, Beckett L, Bold RJ . Incidence, mechanism and prognostic value of activated AKT in pancreas cancer. Br J Cancer 2003; 89: 2110–2115.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Eccles SA . The role of c-erbB-2/HER2/neu in breast cancer progression and metastasis. J Mammary Gland Biol Neoplasia 2001; 6: 393–406.

    Article  CAS  PubMed  Google Scholar 

  12. Schade B, Lam SHL, Cernea D, Sanguin-Gendreau V, Cardiff RD, Jung BL et al. Distinct ErbB-2-coupled signaling pathways promote mammary tumors with unique pathologic and transcriptional profiles. Cancer Res 2007; 67: 7579–7588.

    Article  CAS  PubMed  Google Scholar 

  13. Press MF, Cordon-Cardo C, Slamon DJ . Expression of the HER-2/neu proto-oncogene in normal human adult and fetal tissues. Oncogene 1990; 5: 953–962.

    CAS  PubMed  Google Scholar 

  14. Slamon DJ, Leyland-Jones B, Shak S, Fuchs H, Paton V, Bajamonde A et al. Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2. N Engl J Med 2001; 344: 783–792.

    Article  CAS  PubMed  Google Scholar 

  15. Piccart-Gebhart MJ, Procter M, Leyland-Jones B, Goldhirsch A, Untch M, Smith I et al. Trastuzumab after adjuvant chemotherapy in HER2-positive breast cancer. N Engl J Med 2005; 353: 1659–1672.

    Article  CAS  PubMed  Google Scholar 

  16. Joensuu H, Kellokumpu-Lehtinen PL, Bono P, Alanko T, Kataja V, Asola R et al. Adjuvant docetaxel or vinorelbine with or without Trastuzumab for breast cancer. N Engl J Med 2006; 354: 809–820.

    Article  CAS  PubMed  Google Scholar 

  17. Kreitman RJ, Pastan I . Immunotoxins for targeted cancer therapy. Adv Drug Deliv Rev 1998; 31: 53–88.

    Article  CAS  PubMed  Google Scholar 

  18. Pastan I, Hassan R, FitzGerald DJ, Kreitman RJ . Immunotoxin therapy of cancer. Nat Rev Cancer 2006; 6: 559–565.

    Article  CAS  PubMed  Google Scholar 

  19. Posey JA, Khazaeli MB, Bookman MA, Nowrouzi A, Grizzle WE, Thornton J et al. A phase I trial of the single-chain immunotoxin SGN-10 (BR96 sFv-PE40) in patients with advanced solid tumors. Clin Cancer Res 2002; 8: 3092–3099.

    CAS  PubMed  Google Scholar 

  20. Jin FS, Youle RJ, Johnson VG, Shiloach J, Fass R, Longo DL et al. Suppression of the immune response to immunotoxins with anti-CD4 monoclonal antibodies. J Immunol 1991; 146: 1806–1811.

    CAS  PubMed  Google Scholar 

  21. Fox E, Hwang JJ, Rizvi N, Sing A, Soho C, Marshall JL et al. Phase I study of the combination of the immunotoxin SGN-10 (BR96sFv-PE40) with docetaxel (DOC) in patients (pts) with solid tumors expressing Lewis-Y. Proc Am Soc Clin Oncol 2003; 222 (abstr 891).

  22. Hassan R, Williams-Gould J, Watson T, Pai-Scherf L, Pastan I . Pretreatment with rituximab does not inhibit the human immune response against the immunogenic protein LMB-1. Clin Cancer Res 2004; 10: 16–18.

    Article  CAS  PubMed  Google Scholar 

  23. Yang AG, Chen SY . A new class of antigen-specific killer cells. Nat Biotechnol 1997; 15: 46–51.

    Article  CAS  PubMed  Google Scholar 

  24. Chen SY, Yang AG, Chen JD, Kute T, King CR, Collier J et al. Potent antitumour activity of a new class of tumour-specific killer cells. Nature 1997; 385: 78–80.

    Article  CAS  PubMed  Google Scholar 

  25. Huang Q, Zhang XW, Wang H, Yan B, Dewhirst M, Li CY . A novel conditionally replicative adenovirus vector targeting telomerase-positive tumor cells. Clin Cancer Res 2004; 10: 1439–1445.

    Article  CAS  PubMed  Google Scholar 

  26. Livak KJ, Schmittgen TD . Analysis of relative gene expression data using real-time quantitative PCR and the 2−CT method. Methods 2001; 25: 402–408.

    Article  CAS  PubMed  Google Scholar 

  27. Fleige S, Walf V, Huch S, Prgomet C, Sehm J, Pfaffl MW . Comparison of relative mRNA quantification models and the impact of RNA integrity in quantitative real-time RT-PCR. Biotechnol Lett 2006; 28: 1601–1613.

    Article  CAS  PubMed  Google Scholar 

  28. Siegall CB, Chaudhary VK, FitzGerald DJ, Pastan I . Functional analysis of domains II, Ib, and III of Pseudomonas exotoxin. J Biol Chem 1989; 264: 14256–14261.

    CAS  PubMed  Google Scholar 

  29. Jinno Y, Ogata M, Chaudhary VK, Willingham MC, Adhya S, Fitagerald D et al. Domain II mutants of Pseudomonas exotoxin deficient in translocation. J Biol Chem 1989; 264: 15953–15959.

    CAS  PubMed  Google Scholar 

  30. Hwang J, Fitzgerald DJ, Adhya S, Pastan I . Functional domains of Pseudomonas exotoxin identified by deletion analysis of the gene expressed in E. coli. Cell 1987; 48: 129–136.

    Article  CAS  PubMed  Google Scholar 

  31. Houchins JP . Immunotoxin-induced apoptosis. Stem cell 2000; 18: 384–385.

    Article  CAS  Google Scholar 

  32. Jørgensen R, Merrill AR, Yates SP, Marquez VE, Schwan AL, Boesen T et al. Exotoxin A-eEF2 complex structure indicates ADP ribosylation by ribosome mimicry. Nature 2005; 436: 979–984.

    Article  PubMed  Google Scholar 

  33. Gallant CV, Raivio TL, Olson JC, Woods DE, Storey DG . Pseudomonas aeruginosa cystic fibrosis clinical isolates produce exotoxin A with altered ADP-ribosyltransferase activity and cytotoxicity. Microbiology 2000; 146: 1891–1899.

    Article  CAS  PubMed  Google Scholar 

  34. Palmer DH, Mautner V, Mirza D, Oliff S, Gerritsen W, van der Sijp JR et al. Virus-directed enzyme prodrug therapy: intratumoral administration of a replication-deficient adenovirus encoding nitroreductase to patients with resectable liver cancer. J Clin Oncol 2004; 22: 1546–1552.

    Article  CAS  PubMed  Google Scholar 

  35. Von Der Thüsen H, Kuiper J, Fekkes ML, De Vos P, Van Berkel TJ, Biessen EA . Attenuation of atherogenesis by systemic and local adenovirus-mediated gene transfer of interleukin-10 in LDLr−/− mice. FASEB J 2001; 15: 2730–2732.

    Article  PubMed  Google Scholar 

  36. Wang Y, Yang Z, Liu S, Kon T, Krol A, Li CY et al. Characterisation of systemic dissemination of nonreplicating adenoviral vectors from tumours in local gene delivery. Br J Cancer 2005; 92: 1414–1420.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Ambriovic-Ristov A, Gabrilovac J, Cimbora-Zovko T, Osmak M . Increased adenoviral transduction efficacy in human laryngeal carcinoma cells resistant to cisplatin is associated with increased expression of integrin αvβ3 and coxsackie adenovirus receptor. Int J Cancer 2004; 110: 660–667.

    CAS  PubMed  Google Scholar 

  38. Hemminki A, Kanerva A, Liu B, Wang M, Alvarez RD, Siegal GP et al. Modulation of coxsackie-adenovirus receptor expression for increased adenoviral transgene expression. Cancer Res 2003; 63: 847–853.

    CAS  PubMed  Google Scholar 

  39. Shieh GS, Shiau AL, Yo YT, Lin PR, Chang CC, Tzai TS et al. Low-dose etoposide enhances telomerase-dependent adenovirus-mediated cytosine deaminase gene therapy through augmentation of adenoviral infection and transgene expression in a syngeneic bladder tumor model. Cancer Res 2006; 66: 9957–9966.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr Siyi Chen for the provision of e23(scFv)-PE40-bearing plasmids and Dr Aǹgang Yang for the provision of SK-BR-3 and SGC-7901cell lines. We are grateful for the technical assistant by Fang Wei, Xiafang Chen, Miaoying Yi, Yuhua Tian, Ping Xu, Kuangchen Xie and Yuanshan Lu in First People's Hospital, Shanghai Jiaotong University, for the statistic analyses by Yongbing Xiang and Yuting Tan in the Cancer Institute of Shanghai Jiaotong University. This work was supported by Grant of National Basic Research Project of China (2004CB518804), National Natural Science Foundation for Outstanding Youth (30325043, 30428015).

Author information

Author notes

  1. X Liu, J Wu & S Zhang

    Present address: Current address: Department of Ophthalmology, EYE & ENT Hospital of Fudan University, Shanghai 200032, China,

Authors and Affiliations

  1. Experimental Center, First People's Hospital, Shanghai Jiaotong University, Shanghai, China

    X Liu, J Wu, S Zhang & Q Huang

  2. Department of Radiation Oncology, University of Colorado Health Sciences Center, Aurora, CO, USA

    C Li

  3. National Laboratory for Oncogene and Related Genes Research, Cancer Institute of Shanghai Jiaotong University, Shanghai, China

    Q Huang

Authors
  1. X Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Q Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to C Li or Q Huang.

Additional information

Supplementary Information accompanies the paper on Cancer Gene Therapy website (http://www.nature.com/cgt)

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liu, X., Wu, J., Zhang, S. et al. Novel strategies to augment genetically delivered immunotoxin molecular therapy for cancer therapy. Cancer Gene Ther 16, 861–872 (2009). https://doi.org/10.1038/cgt.2009.30

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/cgt.2009.30

Keywords

Search

Advanced search

Quick links