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

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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.

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Acknowledgments

We thank Ms. Nana Kawaguchi, Ms. Kumi Kodama, Ms. Aya Torisawa, Ms. Keiko Shimoura, and Ms. Maiko Yamada of the Department of Pharmacoepidemiology, Kyoto University, for technical assistance with cell culturing and animal care. This study was supported by a grant-in-aid for Young Scientists (A) (grant no. 23680089) from the Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Department of Pharmacoepidemiology, Graduate School of Medicine and Public Health, Kyoto University, Yoshidakonoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan

    Megumi Kawamoto, Masayuki Kohno, Tomohisa Horibe & Koji Kawakami

Authors
  1. Megumi Kawamoto
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  2. Masayuki Kohno
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  3. Tomohisa Horibe
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  4. Koji Kawakami
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Corresponding author

Correspondence to Koji Kawakami.

Electronic supplementary material

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280_2013_2074_MOESM1_ESM.ppt

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

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)

Supplementary material 3 (DOC 41 kb)

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Kawamoto, M., Kohno, M., Horibe, T. et al. Immunogenicity and toxicity of transferrin receptor-targeted hybrid peptide as a potent anticancer agent. Cancer Chemother Pharmacol 71, 799–807 (2013). https://doi.org/10.1007/s00280-013-2074-4

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  • DOI: https://doi.org/10.1007/s00280-013-2074-4

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