novel cytotoxic vectors based on adeno-associated virus新颖的基于腺相关病毒的细胞毒性向量.pdfVIP

Toxins 2010, 2, 2754-2768; doi:10.3390/toxins2122754 OPEN ACCESS toxins ISSN 2072-6651 /journal/toxins Article Novel Cytotoxic Vectors Based on Adeno-Associated Virus Johannes Kohlschütter †, Stefan Michelfelder † and Martin Trepel * Hubertus Wald Cancer Center, Department of Oncology and Hematology, University Medical Center Hamburg-Eppendorf, Martinistr. 52, 20246 Hamburg, Germany; E-Mails: johannes.kohlschuetter@onlin e.de (J.K.); s.michelfelder@uke.de (S.M.) * Author to whom correspondence should be addressed; E-Mail: m.trepel@uke.de; Tel.: +49-40-741051980; Fax: +49-40-741057187. † These authors contributed equally to this work. Received: 11 November 2010; in revised form: 29 November 2010 / Accepted: 30 November 2010 / Published: 1 December 2010 Abstract: Vectors based on adeno-associated virus (AAV) are promising tools for gene therapy. The production of strongly toxic vectors, for example for cancer-directed gene transfer, is often unfeasible due to uncontrolled expression of toxic genes in vector-producing cells. Using an approach based on transcriptional repression, we have created novel AAV vectors carrying the genes coding for diphtheria toxin A (DTA) and the pro-apoptotic PUMA protein. The DTA vector had a significant toxic effect on a panel of tumor cell lines, and abrogation of protein synthesis could be shown. The PUMA vector had a toxic effect on HeLa and RPMI 8226 cells, and