Abstract
We performed a pilot trial in refractory cancer patients to investigate the feasibility of intratumoral injection of TAPET-CD, an attenuated Salmonella bacterium expressing the E. coli cytosine deaminase gene. A total of three patients received three dose levels of TAPET-CD (3 × 106–3 × 107 CFU/m2) via intratumoral injection once every 28 days as long as progression of disease or intolerable toxicity was not observed. From days 4 to 14 of each 28 day cycle, patients also received 5-fluorocytosine (5-FC) at a dose of 100 mg/kg/day p.o. divided three times daily. Six cycles of treatment were administered. No significant adverse events clearly attributable to TAPET-CD were demonstrated. Two patients had intratumor evidence of bacterial colonization with TAPET-CD, which persisted for at least 15 days after initial injection. Conversion of 5-FC to 5-fluorouracil (5-FU) as a result of cytosine deaminase expression was demonstrated in these two patients. The tumor to plasma ratio of 5-FU for these two colonized patients was 3.0, demonstrating significantly increased levels of 5-FU at the site of TAPET-CD colonization and insignificant systemic spread of the bacteria. In contrast, the tumor to plasma ratio of 5-FU of the patient who did not show colonization of TAPET-CD was less than 1.0. These results support the principle that a Salmonella bacterium can be utilized as a delivery vehicle of the cytosine deaminase gene to malignant tissue and that the delivered gene is functional (i.e. able to convert 5-FC to 5-FU) at doses at or below 3 × 107 CFU/m2.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
References
Mose JR, Mose G . Onkolyseversuche mit apathogenen anaeroben Sporenbildern am Ehrlich Tumor des Maus. Z Krebsforsch. 1959;63:63–74.
Mose JR, Mose G . Oncolysis by clostridia. I. Activity of Clostridium butyricum (M-55) and other nonpathogenic clostridia against the Ehrlich carcinoma. Cancer Res. 1964;24:212–216.
Thiele EH, Arison R, Boxer GE . Oncolysis by clostridia. III. Effects of clostridia and chemotherapeutic agents on rodent tumors. Cancer Res. 1964;2:222–233.
Engelbart K, Gericke D . Oncolysis by clostridia V. Transplanted tumors of the hamster. Cancer Res. 1954;24:239–243.
Carey RW, Holland JF, Whang HY, et al. Clostridial oncolysis in man. Eur J Cancer. 1967;3:37–46.
Heppner F, Mose J . The liquefaction (oncolysis) of malignant gliomas by a non-pathogenic clostridium. Acta Neurol. 1978;12:123–125.
Heppner F, Mose J, Ascher PW, Walter G . Oncolysis of malignant gliomas of the brain. The 13th International Congress on Chemotherapy, 1983: vol 226, 38–45.
Giel CP . Abscess formation in pheochromocytoma. N Engl J Med. 1954;251:980–982.
Simmers TA, Mijnhout G, Van Meyel JJ . Salmonellosis: an usual complication of hepatocellular carcinoma. Scand J Gastroenterol. 1997;32:1180–1182.
Noguerado A, Cabanyes J, Vivancos J, et al. Abscess caused by Salmonella enteritidis within a glioblastoma multiforme. J Infect. 1987;15:61–63.
Black PH, Kunz LJ, Swartz MN . Salmonellosis: a review of some unusual aspects. N Engl J Med. 1960;262:921–926.
Rodriguez RE, Valero V, Watanakunakom C . Salmonella focal intracranial infections: review of the world literature (1884–1984) and report of an usual case. Rev Infect Dis. 1986;8:31–41.
Grahm FO, Coleman PN . Infection of a secondary carcinoma by Salmonella montevideo. BMJ. 1952;1:1116.
Luo X, Ittensohn M, Liu Y, et al. Genetically modified Salmonella typhimurium inhibited growth of primary tumors and metastases. Proc Am Assoc Cancer Res. 1999;40:87.
Low K, Ittensohn M, Lin S, et al. VNP20009, a genetically modified Salmonella typhimurium for treatment of solid tumors. Proc Am Assoc Cancer Res. 1999;40:87.
Low KB, Ittensohn M, Le T, et al. Lipid A mutant salmonella with suppressed virulence and TNFalpha induction retain tumor-targeting in vivo. Nat Biotechnol. 1999;17:37–41.
Pawelek JM, Low KB, Bermudes D . Tumor-targeted salmonella as a novel anticancer vector. Cancer Res. 1997;57:4537–4544.
Khan SA, Everest P, Servos S . A lethal role for lipid A in salmonella infection. Mol Microbiol. 1998;29:571–579.
Lee KC, Zheng LM, Margitich D, Almassian B, King I . Evaluation of the acute and subchronic toxic effects in mice, rats, and monkeys of the genetically engineered and Escherichia coli cytosine deaminase gene-incorporated Salmonella strain. TAPET-CD, being developed as an anti-tumor agent. Int J Toxicol. 2001;20:207–217.
Zheng LM, Luo X, Feng M, et al. Tumor amplified protein expression therapy: Salmonella as a tumor-selective protein delivery vector. Oncol Res. 2000;12:127–135.
Luo X, Li Z, Lin S, et al. Anti-tumor effect of VNP20009, an attenuated Salmonella, in murine tumor models. Oncol Res. 2002;12:501–508.
Toso JF, Gill VJ, Hwu P, et al. Phase I study of the intravenous administration of attenuated Salmonella typhimurium to patients with metastatic melanoma. J Clin Oncol. 2002;20:142–152.
Dinarello CA . Proinflammatory and anti-inflammatory cytokines as mediators in the pathogenesis of septic shock. Chest. 1997;112:321S–329S.
King I, Bermudes D, Lin S, et al. Tumor-targeted Salmonella expressing cytosine deaminase as an anticancer agent. Hum Gene Ther. 2002;13:1225–1233.
Deonarain MP, Spooner RA, Epenetos AA . Genetic delivery of enzymes for cancer therapy. Gene Therapy. 1995;2:235–244.
Haskell CM, Fluorouracil. In: In Cancer Treatment. WB Sounds Company. 1995: 116–119.
Austin EA, Huber BE . A first step in the development of gene therapy for colorectal carcinoma: cloning, sequencing and expression of Escherichia coli cytosine deaminase. Mol Pharmacol. 1993;43:380–387.
Liu SC, Minton NP, Giaccia AJ, Brown JM . Anticancer efficacy of systemically delivered anaerobic bacteria as gene therapy vectors targeting tumor hypoxia/necrosis. Gene Therapy. 2002;9:291–296.
Donnenburg MS, Kaper JB . Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector. Infect Immunity. 1991;59:4310–4317.
Platt J, Sodi S, Kelly M, et al. Antitumor effects of genetically engineered Salmonella in combination with radiation. Eur J Cancer. 2000;36:2397–2402.
Khil MS, Kim JH, Mullen CA, et al. Radiosensitization of 5-fluorocytosine of human colorectal carcinoma cells in culture transduced with cytosine deaminase gene. Clin Cancer Res. 1996;2:53–57.
Gabel M, Kim JH, Kolozsvary A, et al. Selective in vivo radiosensitization by 5-fluorouracil of human colorectal carcinoma cells transduced with the E. coli cytosine deaminase gene. Int J Radiat Oncol Biol Phys. 1998;41:883–887.
Lawrence TS, Rehemtulla A, Ng EY . Preferential cytotoxicity of cells transduced with cytosine deaminase compared to bystander cells after treatment with 5-fluorocytosine. Cancer Res. 1998;58:2588–2593.
Consalvo M, Mullen CA, Modesti A, et al. 5-Fluorocytosine-induced eradication of murine adenocarcinomas engineered to express the cytosine deaminase suicide gene requires host immune competence and leaves and efficient memory. J Immunol. 1995;154:5302–5312.
Pierrefite-Carle V, Baque P, Gavelli A, et al. Cytosine deaminase/5-fluorocytosine-based vaccination against liver tumors: evidence of distant bystander effect. J Natl Cancer Inst. 1999;91:2014–2019.
Hermiston T . Fighting fire with fire: attacking the complexity of human tumors with armed therapeutic viruses. Curr Opin Mol Ther. 2002;4:334–342.
Galanis E, Vile R, Russell SJ . Delivery systems intended for in vivo gene therapy of cancer: targeting and replication competent viral vectors. Crit Rev Oncol Hematol. 2001;38:177–192.
Greco O, Dachs GU . Gene directed enzyme/prodrug therapy of cancer: historical appraisal and future prospectives. J Cell Physiol. 2001;187:22–36.
Crystal RG, Hirschowitz E, Lieberman M, et al. Phase I study of direct administration of a replication deficient adenovirus vector containing the E. coli cytosine deaminase gene to metastatic colon carcinoma of the liver in association with the oral administration of the pro-drug 5-fluorocytosine. Hum Gene Ther. 1997;8:985–1001.
Acknowledgements
We acknowledge Brenda Marr and Elizabeth Asilo for their competent and knowledgeable assistance in the preparation of this manuscript.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Nemunaitis, J., Cunningham, C., Senzer, N. et al. Pilot trial of genetically modified, attenuated Salmonella expressing the E. coli cytosine deaminase gene in refractory cancer patients. Cancer Gene Ther 10, 737–744 (2003). https://doi.org/10.1038/sj.cgt.7700634
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.cgt.7700634
Keywords
This article is cited by
-
Intratumoural microbiota: a new frontier in cancer development and therapy
Signal Transduction and Targeted Therapy (2024)
-
Camouflaging attenuated Salmonella by cryo-shocked macrophages for tumor-targeted therapy
Signal Transduction and Targeted Therapy (2024)
-
Intratumoural microbiota: from theory to clinical application
Cell Communication and Signaling (2023)
-
Microbial applications for sustainable space exploration beyond low Earth orbit
npj Microgravity (2023)
-
A Systemic Review on Fitness and Survival of Salmonella in Dynamic Environment and Conceivable Ways of Its Mitigation
Indian Journal of Microbiology (2023)