Skip to main content

Advertisement

SpringerLink
Log in

Tuftsin-based, EGFR-targeting fusion protein and its enediyne-energized analog show high antitumor efficacy associated with CD47 down-regulation

  • Original Article
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Tuftsin (TF) is an immunomodulator tetrapeptide (Thr-Lys-Pro-Arg) that binds to the receptor neuropilin-1 (Nrp1) on the surface of cells. Many reports have described anti-tumor activity of tuftsin to relate with nonspecific activation of the host immune system. Lidamycin (LDM) that displays extremely potent cytotoxicity to cancer cells is composed of an apoprotein (LDP) and an enediyne chromophore (AE). In addition, Ec is an EGFR-targeting oligopeptide. In the present study, LDP was used as protein scaffold and the specific carrier for the highly potent AE. Genetically engineered fusion proteins LDP-TF and Ec-LDP-TF were prepared; then, the enediyne-energized fusion protein Ec-LDM-TF was generated by integration of AE into Ec-LDP-TF. The tuftsin-based fusion proteins LDP-TF and Ec-LDP-TF significantly enhanced the phagocytotic activity of macrophages as compared with LDP (P < 0.05). Ec-LDP-TF effectively bound to tumor cells and macrophages; furthermore, it markedly suppressed the growth of human epidermoid carcinoma A431 xenograft in athymic mice by 84.2 % (P < 0.05) with up-regulated expression of TNF-α and IFN-γ. Ec-LDM-TF further augmented the therapeutic efficacy, inhibiting the growth of A431 xenograft by 90.9 % (P < 0.05); notably, the Ec-LDM-TF caused marked down-regulation of CD47 in A431 cells. Moreover, the best therapeutic effect was recorded in the group of animals treated with the combination of Ec-LDP-TF with Ec-LDM-TF. The results suggest that tuftsin-based, enediyne-energized, and EGFR-targeting fusion proteins exert highly antitumor efficacy with CD47 modulation. Tuftsin-based fusion proteins are potentially useful for treatment of EGFR- and CD47-overexpressing cancers.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

AE:

Active enediyne from lidamycin

BSA:

Bovine serum albumin

CSCs:

Cancer stem cells

DSB:

DNA double-strand breaks

Ec:

The C-loop of epidermal growth factor (22 amino acids of EGF COOH terminal)

Ec-LDM-TF:

The enediyne-energized fusion protein composed of Ec, LDP, TF and AE

Ec-LDP-TF:

The fusion protein composed of Ec, LDP and TF

EGF:

Epidermal growth factor

EGFR:

Epidermal growth factor receptor

ELISA:

Enzyme-linked immunosorbent assay

FACS:

Fluorescence-activated cell sorting

FITC:

Fluorescein isothiocyanate

HER2:

Human epidermal growth factor receptor-2

HPLC:

High-performance liquid chromatography

HRP:

Horseradish peroxidase

IFN:

Interferon

IgG:

Immunoglobulin

LDM:

Lidamycin (composed of LDP and AE)

LDM-TF:

The enediyne-energized fusion protein composed of LDP, TF and AE

LDP:

Lidamycin apoprotein (110 amino acids)

LDP-TF:

The fusion protein composed of LDP and TF

Nrp1:

Receptor neuropilin-1

TF:

Tuftsin (4 amino acids)

TGF-β:

Transforming growth factor beta

TGI:

Tumor growth inhibition

TNF:

Tumor necrosis factor

References

  1. Fridkin M, Najjar VA (1989) Tuftsin: its chemistry, biology, and clinical potential. Crit Rev Biochem Mol Biol 24:1–40

    Article  CAS  PubMed  Google Scholar 

  2. Nishioka K (1979) Anti-tumour effect of the physiological tetrapeptide, tuftsin. Br J Cancer 39:342–345

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  3. Jain S, Amiji M (2012) Tuftsin-modified alginate nanoparticles as a noncondensing macrophage-targeted DNA delivery system. Biomacromolecules 13:1074–1085

    Article  CAS  PubMed  Google Scholar 

  4. Nissen JC, Selwood DL, Tsirka SE (2013) Tuftsin signals through its receptor neuropilin-1 via the transforming growth factor beta pathway. J Neurochem 127:394–402

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Khan A, Khan AA, Dwivedi V, Ahmad MG, Hakeem S, Owais M (2007) Tuftsin augments antitumor efficacy of liposomized etoposide against fibrosarcoma in Swiss albino mice. Mol Med 13:266–276

    CAS  PubMed Central  PubMed  Google Scholar 

  6. Nishioka K, Babcock GF, Phillips JH, Banks RA, Amoscato AA (1983) In vivo and in vitro antitumor activities of tuftsin. Ann N Y Acad Sci 419:234–241

    Article  CAS  PubMed  Google Scholar 

  7. Yuan W, Xia G, Zhao C, Sui C, Ma J (2012) Anti-idiotypic single chain mimicking CA125 linked with tuftsin provides protective immunity against ovarian cancer in mice. Mol Med Rep 5:388–394

    CAS  PubMed  Google Scholar 

  8. Jaiswal S, Chao MP, Majeti R, Weissman IL (2010) Macrophages as mediators of tumor immunosurveillance. Trends Immunol 31:212–219

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. Prud’Homme GJ, Glinka Y (2012) Neuropilins are multifunctional coreceptors involved in tumor initiation, growth, metastasis and immunity. Oncotarget 3:921–939

    PubMed Central  PubMed  Google Scholar 

  10. Shao RG, Zhen YS (1995) Relationship between the molecular composition of C1027, a new macromolecular antibiotic with enediyne chromophore, and its antitumor activity. Yao Xue Xue Bao 30:336–342

    CAS  PubMed  Google Scholar 

  11. Tanaka T, Fukuda-Ishisaka S, Hirama M, Otani T (2001) Solution structures of C-1027 apoprotein and its complex with the aromatized chromophore. J Mol Biol 309:267–283

    Article  CAS  PubMed  Google Scholar 

  12. Cai L, Chen H, Miao Q, Wu S, Shang Y, Zhen Y (2009) Binding capability of the enediyne-associated apoprotein to human tumors and constitution of a ligand oligopeptide-integrated protein. J Biotechnol 144:142–150

    Article  CAS  PubMed  Google Scholar 

  13. Kennedy DR, Gawron LS, Ju J, Liu W, Shen B, Beerman TA (2007) Single chemical modifications of the C-1027 enediyne core, a radiomimetic antitumor drug, affect both drug potency and the role of ataxia-telangiectasia mutated in cellular responses to DNA double-strand breaks. Cancer Res 67:773–781

    Article  CAS  PubMed  Google Scholar 

  14. Normanno N, De Luca A, Bianco C, Strizzi L, Mancino M, Maiello MR, Carotenuto A, De Feo G, Caponigro F, Salomon DS (2006) Epidermal growth factor receptor (EGFR) signaling in cancer. Gene 366:2–16

    Article  CAS  PubMed  Google Scholar 

  15. Kokai Y, Myers JN, Wada T, Brown VI, LeVea CM, Davis JG, Dobashi K, Greene MI (1989) Synergistic interaction of p185c-neu and the EGF receptor leads to transformation of rodent fibroblasts. Cell 58:287–292

    Article  CAS  PubMed  Google Scholar 

  16. Rathore D, Nayak SK, Batra JK (1996) Expression of ribonucleolytic toxin restrictocin in Escherichia coli: purification and characterization. FEBS Lett 392:259–262

    Article  CAS  PubMed  Google Scholar 

  17. Miao QF, Liu XY, Shang BY, Ouyang ZG, Zhen YS (2007) An enediyne-energized single-domain antibody-containing fusion protein shows potent antitumor activity. Anticancer Drugs 18:127–137

    Article  CAS  PubMed  Google Scholar 

  18. Zhong G, Zhang S, Li Y, Liu X, Gao R, Miao Q, Zhen Y (2010) A tandem scFv-based fusion protein and its enediyne-energized analogue show intensified therapeutic efficacy against lung carcinoma xenograft in athymic mice. Cancer Lett 295:124–133

    Article  CAS  PubMed  Google Scholar 

  19. Guo XF, Zhu XF, Shang Y, Zhang SH, Zhen YS (2010) A bispecific enediyne-energized fusion protein containing ligand-based and antibody-based oligopeptides against epidermal growth factor receptor and human epidermal growth factor receptor 2 shows potent antitumor activity. Clin Cancer Res 16:2085–2094

    Article  CAS  PubMed  Google Scholar 

  20. Roh YJ, Park YG, Kang S, Kim SY, Moon JI (2012) Effects of AFP-172 on COX-2-induced angiogenic activities on human umbilical vein endothelial cells. Graefes Arch Clin Exp Ophthalmol 250:1765–1775

    Article  CAS  PubMed  Google Scholar 

  21. Willingham SB, Volkmer JP, Gentles AJ, Sahoo D, Dalerba P, Mitra SS, Wang J, Contreras-Trujillo H, Martin R, Cohen JD, Lovelace P, Scheeren FA, Chao MP, Weiskopf K, Tang C, Volkmer AK, Naik TJ, Storm TA, Mosley AR, Edris B, Schmid SM, Sun CK, Chua MS, Murillo O, Rajendran P, Cha AC, Chin RK, Kim D, Adorno M, Raveh T, Tseng D, Jaiswal S, Enger PO, Steinberg GK, Li G, So SK, Majeti R, Harsh GR, van de Rijn M, Teng NN, Sunwoo JB, Alizadeh AA, Clarke MF, Weissman IL (2012) The CD47-signal regulatory protein alpha (SIRPa) interaction is a therapeutic target for human solid tumors. Proc Natl Acad Sci USA 109:6662–6667

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  22. Sheng W, Shang Y, Miao Q, Li Y, Zhen Y (2012) Antitumor efficacy of the scFv-based fusion protein and its enediyne-energized analogue directed against epidermal growth factor receptor. Anticancer Drugs 23:406–416

    Article  CAS  PubMed  Google Scholar 

  23. Jiang WG, Lu XA, Shang BY, Fu Y, Zhang SH, Zhou D, Li L, Li Y, Luo Y, Zhen YS (2013) Genetically engineered endostatin-lidamycin fusion proteins effectively inhibit tumor growth and metastasis. BMC Cancer 13:479

    Article  PubMed Central  PubMed  Google Scholar 

  24. Liu X, Guo J, Han S, Yao L, Chen A, Yang Q, Bo H, Xu P, Yin J, Zhang Z (2012) Enhanced immune response induced by a potential influenza A vaccine based on branched M2e polypeptides linked to tuftsin. Vaccine 30:6527–6533

    Article  CAS  PubMed  Google Scholar 

  25. Wu M, Nissen JC, Chen EI, Tsirka SE (2012) Tuftsin promotes an anti-inflammatory switch and attenuates symptoms in experimental autoimmune encephalomyelitis. PLoS ONE 7:e34933

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  26. Siemion IZ, Kluczyk A (1999) Tuftsin: on the 30-year anniversary of Victor Najjar’s discovery. Peptides 20:645–674

    Article  CAS  PubMed  Google Scholar 

  27. Swardfager W, Lanctot K, Rothenburg L, Wong A, Cappell J, Herrmann N (2010) A meta-analysis of cytokines in Alzheimer’s disease. Biol Psychiatry 68:930–941

    Article  CAS  PubMed  Google Scholar 

  28. Schoenborn JR, Wilson CB (2007) Regulation of interferon-gamma during innate and adaptive immune responses. Adv Immunol 96:41–101

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors acknowledge the National High Technology Research and Development Program (No. 2012AA02A301) and “Significant New Drug Development” Major Science and Technology Development Projects of China (No. 2013ZX09102064).

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, No. 1, Tiantan Xili, Beijing, 100050, China

    Wen-Juan Liu, Xiu-Jun Liu, Liang Li, Yi Li, Sheng-Hua Zhang & Yong-Su Zhen

  2. Department of Radiation Oncology, Shandong Cancer Hospital and Institute, Jinan, 250117, Shandong, China

    Wen-Juan Liu

Authors
  1. Wen-Juan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiu-Jun Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Liang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yi Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sheng-Hua Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yong-Su Zhen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yong-Su Zhen.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 421 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, WJ., Liu, XJ., Li, L. et al. Tuftsin-based, EGFR-targeting fusion protein and its enediyne-energized analog show high antitumor efficacy associated with CD47 down-regulation. Cancer Immunol Immunother 63, 1261–1272 (2014). https://doi.org/10.1007/s00262-014-1604-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00262-014-1604-1

Keywords

Search

Navigation