HER2-targeted recombinant protein immuno-caspase-6 effectively induces apoptosis in HER2-overexpressing GBM cells in vitro and in vivo

Zhang,Leiming; Ren,Junlin; Zhang,Hangyu; Cheng,Gang; Xu,Yanming; Yang,Shuangwu; Dong,Chao; Fang,Dandong; Zhang,Jianning; Yang,Angang

doi:10.3892/or.2016.5088

HER2-targeted recombinant protein immuno-caspase-6 effectively induces apoptosis in HER2-overexpressing GBM cells in vitro and in vivo

Authors:
- Leiming Zhang
- Junlin Ren
- Hangyu Zhang
- Gang Cheng
- Yanming Xu
- Shuangwu Yang
- Chao Dong
- Dandong Fang
- Jianning Zhang
- Angang Yang
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Affiliations: Department of Neurosurgery, PLA Navy General Hospital, Beijing 100048, P.R. China, Department of Clinical Laboratory, PLA Navy General Hospital, Beijing 100048, P.R. China, Department of Hepatobiliary Surgery, Xijing Institute of Clinical Neuroscience, Xijing Hospital, Xi'an, Shaanxi 710032, P.R. China, State Key Laboratory of Cancer Biology, Department of Immunology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Neurosurgery, Xijing Institute of Clinical Neuroscience, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China
Published online on: September 12, 2016 https://doi.org/10.3892/or.2016.5088
Pages: 2689-2696

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Abstract

Glioblastoma multiforme (GBM), which is associated with a high rate of morbidity and mortality, is among the most malignant and treatment-refractory neoplasms in human adults. As GBM is highly resistant to conventional therapies, immunotherapies are a promising treatment candidate. HER2 is an attractive target for GBM immunotherapy, as its expression is highly associated with various types of GBM. We previously reported that a novel HER2-targeted recombinant protein e23sFv-Fdt-casp6 has an antitumor effect on HER2-positive gastric cancer cells. In this study, we established a genetically modified Chinese hamster ovary cell line, which produced and secreted e23sFv-Fdt-casp6 proteins. Following specific binding to and internalization into HER2-overexpressing tumor cells, the e23sFv-Fdt-casp6 protein induced tumor cell apoptosis and inhibited the proliferation of HER2-overexpressing A172 and U251MG cells in vitro, but not in U87MG cells with undetectable HER2. The e23sFv-Fdt-casp6 gene was introduced into severe combined immunodeficient mice bearing human glioblastoma xenografts by using intramuscular injections of a liposome-encapsulated vector. The recombinant protein e23sFv-Fdt-casp6 specifically targeted tumor cells and induced apoptosis, thereby leading to potent inhibition of tumor growth and prolonged the survival time of tumor-bearing mice. We concluded that e23sFv‑Fdt‑casp6 represents a promising HER2-targeted treatment option for human gliomas.

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1	Wen PY and Kesari S: Malignant gliomas in adults. N Engl J Med. 359:492–507. 2008. View Article : Google Scholar : PubMed/NCBI
2	Furnari FB, Fenton T, Bachoo RM, Mukasa A, Stommel JM, Stegh A, Hahn WC, Ligon KL, Louis DN, Brennan C, et al: Malignant astrocytic glioma: Genetics, biology, and paths to treatment. Genes Dev. 21:2683–2710. 2007. View Article : Google Scholar : PubMed/NCBI
3	Ohgaki H and Kleihues P: Genetic pathways to primary and secondary glioblastoma. Am J Pathol. 170:1445–1453. 2007. View Article : Google Scholar : PubMed/NCBI
4	Lupu R, Colomer R, Kannan B and Lippman ME: Characterization of a growth factor that binds exclusively to the erbB-2 receptor and induces cellular responses. Proc Natl Acad Sci USA. 89:2287–2291. 1992. View Article : Google Scholar : PubMed/NCBI
5	Koka V, Potti A, Forseen SE, Pervez H, Fraiman GN, Koch M and Levitt R: Role of Her-2/neu overexpression and clinical determinants of early mortality in glioblastoma multiforme. Am J Clin Oncol. 26:332–335. 2003. View Article : Google Scholar : PubMed/NCBI
6	Ahmed N, Salsman VS, Kew Y, Shaffer D, Powell S, Zhang YJ, Grossman RG, Heslop HE and Gottschalk S: HER2-specific T cells target primary glioblastoma stem cells and induce regression of autologous experimental tumors. Clin Cancer Res. 16:474–485. 2010. View Article : Google Scholar : PubMed/NCBI
7	Liu G, Ying H, Zeng G, Wheeler CJ, Black KL and Yu JS: HER-2, gp100, and MAGE-1 are expressed in human glioblastoma and recognized by cytotoxic T cells. Cancer Res. 64:4980–4986. 2004. View Article : Google Scholar : PubMed/NCBI
8	Bian XW, Shi JQ and Liu FX: Pathologic significance of proliferative activity and oncoprotein expression in astrocytic tumors. Anal Quant Cytol Histol. 22:429–437. 2000.
9	Forseen SE, Potti A, Koka V, Koch M, Fraiman G and Levitt R: Identification and relationship of HER-2/neu overexpression to short-term mortality in primary malignant brain tumors. Anticancer Res. 22:1599–1602. 2002.PubMed/NCBI
10	Mineo JF, Bordron A, Quintin-Roué I, Loisel S, Ster KL, Buhé V, Lagarde N and Berthou C: Recombinant humanised anti-HER2/neu antibody (Herceptin) induces cellular death of glioblastomas. Br J Cancer. 91:1195–1199. 2004.PubMed/NCBI
11	Press MF, Cordon-Cardo C and Slamon DJ: Expression of the HER-2/neu proto-oncogene in normal human adult and fetal tissues. Oncogene. 5:953–962. 1990.PubMed/NCBI
12	Bold RJ, Termuhlen PM and McConkey DJ: Apoptosis, cancer and cancer therapy. Surg Oncol. 6:133–142. 1997. View Article : Google Scholar
13	Riedl SJ and Shi Y: Molecular mechanisms of caspase regulation during apoptosis. Nat Rev Mol Cell Biol. 5:897–907. 2004. View Article : Google Scholar : PubMed/NCBI
14	Srinivasula SM, Ahmad M, MacFarlane M, Luo Z, Huang Z, Fernandes-Alnemri T and Alnemri ES: Generation of constitutively active recombinant caspases-3 and -6 by rearrangement of their subunits. J Biol Chem. 273:10107–10111. 1998. View Article : Google Scholar : PubMed/NCBI
15	Komata T, Kondo Y, Kanzawa T, Hirohata S, Koga S, Sumiyoshi H, Srinivasula SM, Barna BP, Germano IM, Takakura M, et al: Treatment of malignant glioma cells with the transfer of constitutively active caspase-6 using the human telomerase catalytic subunit (human telomerase reverse transcriptase) gene promoter. Cancer Res. 61:5796–5802. 2001.PubMed/NCBI
16	Ren JL, Meng YL, Hu B, Jia LT, Zhang R, Xu YM, Xie QS, Zhang YQ, Jin BQ, Chen SY, et al: The effect of direct translocation across endosomes on the cytotoxicity of the recombinant protein e23sFv-Fdt-casp6 to HER2 positive gastric cancer cells. Biomaterials. 32:7641–7650. 2011. View Article : Google Scholar : PubMed/NCBI
17	Wang T, Zhao J, Ren JL, Zhang L, Wen WH, Zhang R, Qin WW, Jia LT, Yao LB, Zhang YQ, et al: Recombinant immunoproapoptotic proteins with furin site can translocate and kill HER2-positive cancer cells. Cancer Res. 67:11830–11839. 2007. View Article : Google Scholar : PubMed/NCBI
18	Omasa T, Onitsuka M and Kim WD: Cell engineering and cultivation of chinese hamster ovary (CHO) cells. Curr Pharm Biotechnol. 11:233–240. 2010. View Article : Google Scholar : PubMed/NCBI
19	Mineo JF, Bordron A, Quintin-Roué I, Maurage CA, Buhé V, Loisel S, Dubois F, Blond S and Berthou C: Increasing of HER2 membranar density in human glioblastoma U251MG cell line established in a new nude mice model. J Neurooncol. 76:249–255. 2006. View Article : Google Scholar
20	Vajkoczy P, Schilling L, Ullrich A, Schmiedek P and Menger MD: Characterization of angiogenesis and microcirculation of high-grade glioma: An intravital multifluorescence microscopic approach in the athymic nude mouse. J Cereb Blood Flow Metab. 18:510–520. 1998. View Article : Google Scholar : PubMed/NCBI
21	Hegde M, Corder A, Chow KK, Mukherjee M, Ashoori A, Kew Y, Zhang YJ, Baskin DS, Merchant FA, Brawley VS, et al: Combinational targeting offsets antigen escape and enhances effector functions of adoptively transferred T cells in glioblastoma. Mol Ther. 21:2087–2101. 2013. View Article : Google Scholar : PubMed/NCBI
22	Akiyama Y, Oshita C, Kume A, Iizuka A, Miyata H, Komiyama M, Ashizawa T, Yagoto M, Abe Y, Mitsuya K, et al: α-type-1 polarized dendritic cell-based vaccination in recurrent high-grade glioma: A phase I clinical trial. BMC Cancer. 12:6232012. View Article : Google Scholar
23	Phuphanich S, Wheeler CJ, Rudnick JD, Mazer M, Wang H, Nuño MA, Richardson JE, Fan X, Ji J, Chu RM, et al: Phase I trial of a multi-epitope-pulsed dendritic cell vaccine for patients with newly diagnosed glioblastoma. Cancer Immunol Immunother. 62:125–135. 2013. View Article : Google Scholar :
24	Sasaki S, Takeshita F, Xin KQ, Ishii N and Okuda K: Adjuvant formulations and delivery systems for DNA vaccines. Methods. 31:243–254. 2003. View Article : Google Scholar : PubMed/NCBI
25	Robinson HL and Pertmer TM: DNA vaccines for viral infections: Basic studies and applications. Adv Virus Res. 55:1–74. 2000. View Article : Google Scholar : PubMed/NCBI
26	Sun Y, Hu YH, Liu CS and Sun L: Construction and analysis of an experimental Streptococcus iniae DNA vaccine. Vaccine. 28:3905–3912. 2010. View Article : Google Scholar : PubMed/NCBI
27	Kim D, Hung CF, Wu TC and Park YM: DNA vaccine with α-galactosylceramide at prime phase enhances anti-tumor immunity after boosting with antigen-expressing dendritic cells. Vaccine. 28:7297–7305. 2010. View Article : Google Scholar : PubMed/NCBI