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Systemic p53 Gene Therapy of Cancer with Immunolipoplexes Targeted by Anti-Transferrin Receptor scFv

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Abstract

Background

A long-standing goal in genetic therapy for cancer is a systemic gene delivery system that selectively targets tumor cells, including metastases. Here we describe a novel cationic immunolipoplex system that shows high in vivo gene transfer efficiency and antitumor efficacy when used for systemic p53 gene therapy of cancer.

Materials and Methods

A cationic immunolipoplex incorporating a biosynthetically lipid-tagged, anti-transferrin receptor single-chain antibody (TfRscFv), was designed to target tumor cells both in vitro and in vivo. A human breast cancer metastasis model was employed to evaluate the in vivo efficacy of systemically administered, TfRscFv-immunolipoplex-mediated, p53 gene therapy in combination with docetaxel.

Results

The TfRscFv-targeting cationic immunolipoplex had a size of 60–100 nm, showed enhanced tumor cell binding, and improved targeted gene delivery and transfection efficiencies, both in vitro and in vivo. The p53 tumor suppressor gene was not only systemically delivered by the immunolipoplex to human tumor xenografts in nude mice but also functionally expressed. In the nude mouse breast cancer metastasis model, the combination of the p53 gene delivered by the systemic administration of the TfRscFv-immunolipoplex and docetaxel resulted in significantly improved efficacy with prolonged survival.

Conclusions

This is the first report using scFv-targeting immunolipoplexes for systemic gene therapy. The TfRscFv has a number of advantages over the transferrin (Tf) molecule itself: (1) scFv has a much smaller size than Tf producing a smaller immunolipoplex giving better penetration into solid tumors; (2) unlike Tf, the scFv is a recombinant protein, not a blood product; (3) large scale production and strict quality control of the recombinant scFv, as well as scFv-immunolipoplex, are feasible. The sensitization of tumors to chemotherapy by this tumor-targeted and efficient p53 gene delivery method could lower the effective dose of the drug, correspondingly lessening the severe side effects, while decreasing the possibility of recurrence. Moreover, this approach is applicable to both primary and recurrent tumors, and more significantly, metastatic disease. The TfRscFv-targeting of cationic immunolipoplexes is a promising method of tumor targeted gene delivery that can be used for systemic gene therapy of cancer with the potential to critically impact the clinical management of cancer.

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Acknowledgments

We are grateful to Dr. David Fitzgerald for providing the plasmid pDFH2T-vecOK containing the TfRscFv cDNA, Dr. John de Kruif for providing the expression vector pLP1, Dr. Arnold Levine for providing the plasmid pBP100, Ms. Tina Wilson and Kelle Reames for assistance with the in vivo animal studies, Dr. Wei-Qun Huang for assistance with Western analysis, Dr. Karen Creswell for assistance with flow cytometry analysis, and Leanne Sleer for assistance with the manuscript. This work was supported in part by NCI grant R01 CA45158 (E. C.) and NCI STTR Phase I grant R41 CA80449 (E. C.), and SynerGene Therapeutics, Inc.

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  1. These authors contributed equally to the study.

Authors and Affiliations

  1. Department of Oncology, Lombardi Cancer Center, The Research Building/E420, Georgetown University Medical Center, 3970 Reservoir Road, NW, 20007, Washington D.C., USA

    Liang Xu, Wen-Hua Tang, Cheng-Cheng Huang, William Alexander, Lai-Man Xiang, Kathleen F. Pirollo, Antonina Rait & Esther H. Chang

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  1. Liang Xu
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  2. Wen-Hua Tang
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  6. Kathleen F. Pirollo
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  7. Antonina Rait
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  8. Esther H. Chang
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Correspondence to Esther H. Chang.

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Xu, L., Tang, WH., Huang, CC. et al. Systemic p53 Gene Therapy of Cancer with Immunolipoplexes Targeted by Anti-Transferrin Receptor scFv. Mol Med 7, 723–734 (2001). https://doi.org/10.1007/BF03401962

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