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TNFerade Biologic: Preclinical toxicology of a novel adenovector with a radiation-inducible promoter, carrying the human tumor necrosis factor alpha gene

Cancer Gene Therapy volume 9, pages 951–957 (2002)Cite this article

Abstract

TNFerade Biologic (TNFerade) is a second-generation (E1-, E3-, and E4-deleted) replication-deficient adenovector carrying the transgene encoding for human tumor necrosis factor alpha (TNFα), regulated by the radiation-sensitive promoter Early Growth Response (Egr-1). We hypothesized that intratumoral injection of TNFerade followed by radiation would result in potentially therapeutic levels of TNFα with minimal toxicity. Three preclinical studies were conducted, the purpose of which was to characterize the toxicity and pharmacokinetics of TNFerade in conjunction with radiation in nude as well as immune-competent (Balb/c) mice. A total of 80 mice in the nude mouse toxicology study, all bearing human squamous cell carcinoma xenografts, 120 mice in the Balb/c study, and 33 nude mice in the pharmacokinetic study were used. Doses ranging from 4×109 to 4×1010 particle units (pu) (4×1011 pu in the Balb/c study) were explored, with and without radiation. In the nude mice studies, TNFerade was injected intratumorally, whereas in the Balb/c study, TNFerade was administered by subcutaneous injection. TNFerade was well tolerated. In the nude mice studies, no significant toxicity occurred in any dose group. In the Balb/c study, 6/40 mice at the top dose (4×1011 pu) were sacrificed in moribund condition (5/20 in the TNFerade+radiation group, 1/20 in the TNFerade alone group). Necropsy showed local necrosis and ulceration at the site of the injection. No deaths or significant toxicity were observed at the lower dose levels (4×109 and 4×1010 pu), indicating a large safety margin for initial studies in humans. The pharmacokinetic study demonstrated high sustained levels of TNFα in the tumor homogenate with no “spillover” to plasma, where TNFα levels were below the level of detection. Radiation increased intratumoral levels of TNFα by a factor of 12 (from 0.998 to 11.55 ng/g). In conclusion, a gene therapy approach with TNFerade, in combination with radiation, represents a potential way to utilize the potent anticancer activity of TNFα without systemic toxicity.

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Acknowledgements

We thank Angela Barber for her competent and knowledgeable assistance in the preparation of this manuscript.

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Authors and Affiliations

  1. GenVec Inc., Gaithersburg, 20878, Maryland, USA

    Henrik Rasmussen, Camilla Rasmussen, Maria Lempicki, Rebecca Durham, Douglas Brough & C Richter King

  2. Department of Radiation and Cellular Oncology, University of Chicago Hospitals, Chicago, Illinois, USA

    Ralph Weichselbaum

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  1. Henrik Rasmussen
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Correspondence to Henrik Rasmussen.

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Rasmussen, H., Rasmussen, C., Lempicki, M. et al. TNFerade Biologic: Preclinical toxicology of a novel adenovector with a radiation-inducible promoter, carrying the human tumor necrosis factor alpha gene. Cancer Gene Ther 9, 951–957 (2002). https://doi.org/10.1038/sj.cgt.7700518

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