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Z Gastroenterol 2004; 42(12): 1363-1370
DOI: 10.1055/s-2004-813310
Originalarbeit

© Karl Demeter Verlag im Georg Thieme Verlag KG Stuttgart · New York

Gentherapie durch einen RGD-modifizierten, hTERT-regulierten, TRAIL-exprimierenden adenoviralen Vektor in malignen Tumoren

Adenoviral Vector Expressing the TRAIL Gene Driven by the hTERT PromoterD. Jacob1 , J. Davis2 , G. Schumacher1 , M. Bahra1 , P. Neuhaus1 , B. Fang2
  • 1Klinik für Transplantations-, Viszeral- und Allgemeinchirurgie, Charité Campus-Virchow, Humboldt-Universität zu Berlin, Deutschland
  • 2Department of Thoracic and Cardiovascular Surgery, The University of Texas M. D. Anderson Cancer Center, and the Program in Gene Therapy and Virology, The University of Texas Graduate School of Biomedical Sciences, Houston, USA
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Publication History

Manuskript eingetroffen: 16.4.2004

Manuskript akzeptiert: 26.5.2004

Publication Date:
09 December 2004 (online)

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Zusammenfassung

Hintergrund: Die Modifikation adenoviraler Vektoren mit einer Arg-Gly-Asp-(RGD-)Sequenz kann Resistenzentwicklungen umgehen und zu einer erhöhten Transfektion der Zelle führen. Wir konstruierten daher einen adenoviralen Vektor mit einer RGD-Sequenz, der das TRAIL-Gen, gesteuert von einem Human-Telomerase-Reverse-Transcriptase-(hTERT-)Promoter, exprimiert, und untersuchten seine zelltötende Aktivität in vitro und in vivo, wozu ein orthotopes Pankreastumormodel in Nacktmäusen etabliert wurde. Material und Methoden: Die Apoptoseinduktion des Vektors Ad/TRAIL-F/RGD wurde in humanen Zelllinien von hepatozellulären Karzinomen (Hep G2, Hep 3b), Pankreaskarzinomen (Panc-1, Capan-1) und Kolonkarzinomen (LOVO, SW 620) untersucht. Die Hemmung der Zellproliferation wurde mit einem XTT-Assay bestimmt, die GFP-Expression und Apoptoseinduktion mittels Durchflusszytometrie sowie TRAIL- und Caspase-8-Expression durch Western Blot-Analysen. In-vivo-Untersuchungen wurden in einem orthotopen Pankreastumormodel an Nu/nu-Nacktmäusen durchgeführt. Ergebnisse: Die Behandlung mit Ad/TRAIL-F/RGD und Ad/gTRAIL zeigte eine signifikant reduzierte Zellproliferation und deutliche Apoptoseinduktion im Vergleich zu den Kontrollgruppen in allen getesteten Zelllinien. Zusätzlich zeigten die mit Ad/TRAIL-F/RGD behandelten Tiere ein signifikant geringeres Tumorwachstum (p < 0,05) als die mit PBS oder einem Kontrollvektor behandelten Tiere. Schlussfolgerung: Unsere Ergebnisse zeigen bei der Verwendung des adenoviralen Vektors Ad/TRAIL-F/RGD in vitro eine signifikante Proliferationshemmung und deutliche Apoptoseinduktion in humanen Tumorzelllinien sowie eine signifikante Tumorwachstumshemmung in orthotop implantierten Pankreastumoren im Pankreasschwanz. Der Einsatz des adenoviralen Vektors Ad/TRAIL-F/RGD bei der Behandlung maligner Tumoren könnte in Zukunft eine Therapieoption darstellen.

Abstract

Background: Resistance can be overcome by modified adenoviral vectors containing an Arg-Gly-Asp (RGD) sequence. We constructed an adenoviral vector with RGD-modified fibers, expressing the TRAIL gene from the human telomerase reverse transcriptase (hTERT) promoter (designated Ad/TRAIL-F/RGD), and evaluated its antitumor activity in vitro and in vivo. Methods: The induction of apoptosis by the new vector Ad/TRAIL-F/RGD was evaluated in human carcinoma cells derived from hepatocellular carcinoma (Hep G2, Hep 3b), pancreatic carcinoma (Panc-1, Capan-1), and colon carcinoma (LOVO, SW 620). Cell viability was measured by the XTT assay and GFP expression and apoptosis induction by fluorescence-activated cell sorting (FACS) and Western blot. In vivo experiments were performed in an orthotopic pancreas tumor model in nu/nu nude mice. Results: Treatment with Ad/TRAIL-F/RGD and Ad/gTRAIL resulted in significantly reduced cell viability in comparison to PBS and Ad/CMV-GFP treatment in all examined human carcinoma cell lines. In addition, mice treated with Ad/TRAIL-F/RGD showed a significantly decreased tumor growth than both control groups. Conclusions: Our results suggest that Ad/TRAIL-F/RGD may become a potent therapeutic agent for the treatment of different human solid carcinomas.

Schlüsselwörter

Apoptose - Gentherapie - TRAIL - RGD - Adenovirus - Gentransfer

Key words

Apoptosis - gene therapy - TRAIL - RGD - adenovirus

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Dr. med. Dietmar Jacob

Klinik für Transplantations-, Viszeral- und Allgemeinchirurgie, Charité Campus-Virchow

Humboldt-Universität zu Berlin

Augustenburger Platz 1

13353 Berlin

Phone: ++ 49/30/4 50-55 20 01

Fax: ++ 49/30/4 50-55 29 00

Email: dietmar.jacob@charite.de

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