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Hapten-directed targeting to single-chain antibody receptors

Cancer Gene Therapy volume 11, pages 380–388 (2004)Cite this article

Abstract

Artificial recombinant receptors may be useful for selectively targeting imaging and therapeutic agents to sites of gene expression. To evaluate this approach, we developed transgenes to express highly on cells a single-chain antibody (scFv) against the hapten 4-ethoxymethylene-2-phenyl-2-oxazoline-5-one (phOx). A phOx enzyme conjugate was created by covalently attaching phOx molecules to polyethylene glycol (PEG)-modified β-glucuronidase. Cells expressing phOx scFv but not control scFv receptors were selectively killed after exposure to ß-glucuronidase derivatized with phOx and PEG (phOx-βG-PEG) and a glucuronide prodrug (p-hydroxy aniline mustard β-D-glucuronide, HAMG) of p-hydroxyaniline mustard. Targeted activation of HAMG produced bystander killing of receptor-negative cells in mixed populations containing as few as 10% phOx-receptor-positive cells. Functional phOx scFv receptors were stably expressed on B16-F1 melanoma tumors in vivo. Treatment of mice bearing established phOx-receptor-positive tumors with phOx-βG-PEG and HAMG significantly (P⩽.0005) suppressed tumor growth as compared with treatment with βG-PEG and HAMG or prodrug alone. phOx was unstable in the serum, suggesting alternative haptens may be more suitable for in vivo applications. Our results show that therapeutic agents can be targeted to artificial hapten receptors in vitro and in vivo. The expression of artificial receptors on target cells may allow preferential delivery of therapeutic or imaging molecules to sites of transgene expression.

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Abbreviations

HAMG:

p-hydroxy aniline mustard β-D-glucuronide

pHAM:

p-hydroxyaniline mustard

phOx:

4-ethoxymethylene-2-phenyl-2-oxazoline-5-one

phOx-ßG-PEG:

ß-glucuronidase derivatized with phOx and polyethylene glycol

scFv:

single-chain antibody

TM:

transmembrane domain

γ1:

H-CH2-CH3 region of human IgG1

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Acknowledgements

We wish to thank Ms Ya-Min Lin of the Institute of Molecular Biology, Academia Sinica for assistance in cell sorting. This study was supported by grants from the National Science Council (NSC91-3112-P-001-026-Y) and Academia Sinica (AS-91-IBMS-3PP), Taipei, Taiwan.

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Authors and Affiliations

  1. School of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, Taiwan

    Tian-Lu Cheng & Chiu-Min Cheng

  2. MedicoGenomic Research Center, Kaohsiung Medical University, Kaohsiung, Taiwan

    Tian-Lu Cheng & Chiu-Min Cheng

  3. NeuroMedical Scientific Center, Buddhist Tzu Chi General Hospital, Hualien, Taiwan

    Kuang-Wen Liao

  4. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan

    Shey-Cherng Tzou, Bing-Mae Chen & Steve R Roffler

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  1. Tian-Lu Cheng
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Correspondence to Steve R Roffler.

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Cheng, TL., Liao, KW., Tzou, SC. et al. Hapten-directed targeting to single-chain antibody receptors. Cancer Gene Ther 11, 380–388 (2004). https://doi.org/10.1038/sj.cgt.7700712

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