Cancer Chemotherapy and Pharmacology

, Volume 71, Issue 3, pp 799–807 | Cite as

Immunogenicity and toxicity of transferrin receptor-targeted hybrid peptide as a potent anticancer agent

  • Megumi Kawamoto
  • Masayuki Kohno
  • Tomohisa Horibe
  • Koji Kawakami
Original Article

Abstract

Purpose

Transferrin receptor (TfR) is a cell membrane-associated glycoprotein involved in the cellular uptake of iron and the regulation of cell growth. Recent studies have shown elevated expression levels of TfR on cancer cells compared with normal cells. We previously designed a TfR-lytic hybrid peptide, which combines the TfR-binding peptide and a lytic peptide, and reported that it bound specifically to TfR and selectively killed cancer cells. Furthermore, the intravenous administration of TfR-lytic peptide in an athymic mouse model significantly inhibited tumor progression. To evaluate the immunogenicity of this peptide as a novel and potent anticancer agent, we investigated whether TfR-lytic hybrid peptide elicits cellular and humoral immune responses to produce antibodies. We also examined the toxicity of this peptide in syngeneic mice.

Methods

We performed hematologic and blood chemistry test and histological analysis and assessed hemolytic activity to check toxicity. To evaluate the immunogenicity, measurement of murine interferon-gamma and detection of TfR-lytic-specific antibody by ELISA were demonstrated.

Results

No T cell immune response or antibodies were detected in the group treated with TfR-lytic hybrid peptide. No hematologic toxicity, except for a decrease in leukocytes, was observed, and no remarkable influence on metabolic parameters and organs (liver, kidney, and spleen) was noted.

Conclusions

Therefore, TfR-lytic hybrid peptide might provide an alternative therapeutic option for patients with cancer.

Keywords

Transferrin receptor Immunogenicity Toxicity Peptide drug Molecular target drug for cancer treatment Immunotoxin 

Supplementary material

280_2013_2074_MOESM1_ESM.ppt (60 kb)
Supplementary Fig. 1 Anti-tumor activity of TfR-lytic hybrid peptide in athymic mouse model in vivo. U251 human glioma cells were implanted subcutaneously into athymic mice. Intravenous injection of either saline (control) or TfR-lytic peptide (3 mg/kg) as indicated by the arrows. Data are expressed as mean ± SD, n = 6 animals in each group. Supplementary material 1 (PPT 60 kb)
280_2013_2074_MOESM2_ESM.ppt (87 kb)
Supplementary Fig. 2 Specific immunoglobulin M for TfR-lytic hybrid peptide or KLH-conjugated TfR-lytic peptide in the serum of treated mice with these peptides. Serum samples were collected on days 12, 19, 26, and 33 in GL261 cells implanting, and (A) used in a serial dilutions, (B) assessed to determine the optical density of serum specimens diluted 1:16. Mean ± SD (n = 3 mice/group). Supplementary material 2 (PPT 87 kb)
280_2013_2074_MOESM3_ESM.doc (41 kb)
Supplementary material 3 (DOC 41 kb)

References

  1. 1.
    Pastan I (1997) Targeted therapy of cancer with recombinant immunotoxins. Biochim Biophys Acta 1333:C1–C6PubMedGoogle Scholar
  2. 2.
    Krietman RJ (2006) Immunotoxins for targeted cancer therapy. AAPS J 8:E532–E551CrossRefGoogle Scholar
  3. 3.
    Rand RW, Kreitman RJ, Patronas N, Varricchio F, Pastan I, Puri RK (2000) Intratumoral administration of recombinant circularly permuted interleukin-4-Pseudomonas exotoxin in patients with high-grade glioma. Clin Cancer Res 6:2157–2165PubMedGoogle Scholar
  4. 4.
    Cintredekin Besudotox Intraparenchymal Study Group (2007) Direct intracerebral delivery of cintredekin besudotox (IL13-PE38QQR) in recurrent malignant glioma: a report by the Cintredekin Besudotox Intraparenchymal Study Group. J Clin Oncol 25:837–844CrossRefGoogle Scholar
  5. 5.
    Foss FM (2000) DAB (389) IL-2 (ONTAK): a novel fusion toxin therapy for lymphoma. Clin Lymphoma 2:110–117Google Scholar
  6. 6.
    Piascik P (1999) FDA approves fusion protein for treatment of lymphoma. J Am Pharm Assoc 39:571–572Google Scholar
  7. 7.
    Frankel AE (2004) Reducing the immune response to immunotoxin commentary re R. Hassan et al. Pretreatment with rituximab does not inhibit the human immune response against the immunogenic protein LMB-1. Clin Cancer Res 10:16–18CrossRefGoogle Scholar
  8. 8.
    Hertler AA, Spitler LE, Frankel AE (1987) Humoral immune response to a ricin A chain immunotoxin in patients with metastatic melanoma. Cancer Drug Deliv 4:245–253PubMedCrossRefGoogle Scholar
  9. 9.
    Kreitman RJ, Wilson WH, White JD, Stetler-Stevenson M, Jaffe ES, Giardina S, Waldmann TA, Pastan I (2000) Phase I trial of recombinant immunotoxin anti-Tac(Fv)-PE38 (LMB-2) in patients with hematologic malignancies. J Clin Oncol 18:1622–1636PubMedGoogle Scholar
  10. 10.
    Neckers LM, Trepel JB (1986) Transferrin receptor expression and the control of cell growth. Cancer Invest 4:461–470PubMedCrossRefGoogle Scholar
  11. 11.
    Habeshaw JA, Lister TA, Stansfeld AG, Greaves MF (1983) Correlation of transferrin receptor expression with histological class and outcome in non-Hodgkin lymphoma. Lancet 1:498–501PubMedCrossRefGoogle Scholar
  12. 12.
    Kondo K, Noguchi M, Mukai Z, Matsuno Y, Sato Y, Shimosato Y, Monden Y (1990) Transferrin receptor expression in adenocarcinoma of the lung as a histopathologic indicator of prognosis. Chest 97:1367–1371PubMedCrossRefGoogle Scholar
  13. 13.
    Walker RA, Day SJ (1986) Transferrin receptor expression in nonmalignant and malignant human breast tissue. J Pathol 148:217–224PubMedCrossRefGoogle Scholar
  14. 14.
    Yang DC, Wang F, Elliott RL, Head JF (2001) Expression of transferrin receptor and ferritin H-chain mRNA are associated with clinical and histopathological prognostic indicators in breast cancer. Anticancer Res 21:541–549PubMedGoogle Scholar
  15. 15.
    Prior R, Reifenberger G, Wechsler W (1990) Transferrin receptor expression in tumours of the human nervous system: relation to tumour type, grading and tumour growth fraction. Virchows Arch A Pathol Anat Histopathol 416:491–496PubMedCrossRefGoogle Scholar
  16. 16.
    Das Gupta A, Shah VI (1990) Correlation of transferrin receptor expression with histologic grade and immunophenotype in chronic lymphocytic leukemia and non-Hodgkin’s lymphoma. Hematol Pathol 4:37–41PubMedGoogle Scholar
  17. 17.
    Kohno M, Horibe T, Haramoto M, Yano Y, Nakajima O, Matsuzaki K, Kawakami K (2011) A novel hybrid peptide targeting EGFR-expressing cancers. Eur J Cancer 47:773–783PubMedCrossRefGoogle Scholar
  18. 18.
    Kawamoto M, Horibe T, Kohno M, Kawakami K (2011) A novel transferrin receptor-targeted hybrid peptide disintegrates cancer cell membrane to induce rapid killing of cancer cells. BMC Cancer 11:359PubMedCrossRefGoogle Scholar
  19. 19.
    Yang L, Horibe T, Kohno M, Haramoto M, Ohara K, Puri RK, Kawakami K (2012) Targeting interleukin-4 receptor α with hybrid peptide for effective cancer therapy. Mol Cancer Ther 1:235–243CrossRefGoogle Scholar
  20. 20.
    Lien S, Lowman HB (2003) Therapeutic peptides. Trends Biotechnol 21:556–562PubMedCrossRefGoogle Scholar
  21. 21.
    Fuessel S, Meye A, Schmitz M, Zastrow S, Linné C, Richter K, Löbel B, Hakenberg OW, Hoelig K, Rieber EP, Wirth MP (2006) Vaccination of hormone-refractory prostate cancer patients with peptide cocktail-loaded dendritic cells: results of a phase I clinical trial. Prostate 66:811–821PubMedCrossRefGoogle Scholar
  22. 22.
    Chromek M, Slamová Z, Bergman P, Kovács L, Podracká L, Ehrén I, Hökfelt T, Gudmundsson GH, Gallo RL, Agerberth B, Brauner A (2006) The antimicrobial peptide cathelicidin protects the urinary tract against invasive bacterial infection. Nat Med 12:636–641PubMedCrossRefGoogle Scholar
  23. 23.
    Kumar P, Wu H, McBride JL, Jung KE, Kim MH, Davidson BL, Lee SK, Shankar P, Manjunath N (2007) Transvascular delivery of small interfering RNA to the central nervous system. Nature 448:39–43PubMedCrossRefGoogle Scholar
  24. 24.
    Lee JH, Engler JA, Collawn JF, Moore BAJ (2001) Receptor mediated uptake of peptides that bind the human transferring receptor. Eur J Biochem 268:2004–2012PubMedCrossRefGoogle Scholar
  25. 25.
    Recht L, Torres CO, Smith TW, Raso V, Griffin TW (1990) Transferrin receptor in normal and neoplastic brain tissue: implications for brain-tumor immunotherapy. J Neurosurg 72:941–945PubMedCrossRefGoogle Scholar
  26. 26.
    Chirasani SR, Markovic DS, Synowitz M, Eichler SA, Wisniewski P, Kaminska B, Otto A, Wanker E, Schäfer M, Chiarugi P, Meier JC, Kettenmann H, Glass R (2009) Transferrin-receptor-mediated iron accumulation controls proliferation and glutamate release in glioma cells. J Mol Med (Berl) 87:153–167CrossRefGoogle Scholar
  27. 27.
    Park JM, Jung JE, Lee BJ (1994) Antimicrobial peptides from the skin of a Korean frog, Rana rugosa. Biochem Biophys Res Commun 205:948–954PubMedCrossRefGoogle Scholar
  28. 28.
    Helmerhorst EJ, Reijnders IM, van ‘t Hof W, Veerman EC, Nieuw Amerongen AV (1999) A critical comparison of the hemolytic and fungicidal activities of cationic antimicrobial peptides. FEBS Lett 449:105–110PubMedCrossRefGoogle Scholar
  29. 29.
    Zagzag D, Salnikow K, Chiriboga L, Yee H, Lan L, Ali MA, Garcia R, Demaria S, Newcomb EW (2005) Downregulation of major histocompatibility complex antigens in invading glioma cells: stealth invasion of the brain. Lab Invest 85:328–341PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Megumi Kawamoto
    • 1
  • Masayuki Kohno
    • 1
  • Tomohisa Horibe
    • 1
  • Koji Kawakami
    • 1
  1. 1.Department of Pharmacoepidemiology, Graduate School of Medicine and Public HealthKyoto UniversitySakyo-ku, KyotoJapan

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