title: Combinatorial anti-angiogenic gene therapy in a human malignant mesothelioma model.
abstract: Anti-angiogenic gene therapy represents a promising strategy for cancer; however, it has rarely been tested in malignant mesothelioma, a highly aggressive tumor associated with asbestos with poor prognosis. In the present study, we investigated whether anti-angiogenic factors such as angiostatin, endostatin and the soluble form of vascular endothelial growth factor receptor 2 (sFlk1) were able to inhibit endothelial cell proliferation via lentivirus-mediated gene transfer into malignant mesothelioma cells in culture. We also assessed whether a dual-agent strategy had greater therapeutic benefit. Human malignant pleural mesothelioma MSTO-211H cells were transduced using lentiviral vectors that individually expressed angiostatin, endostatin and sFlk1 and linked to enhanced green fluorescent protein (EGFP) marker gene expression via an internal ribosome entry site. The lentivirus expressing EGFP alone was used as a control. The resultant cells designated as MSTO-A, MSTO-E, MSTO-F and MSTO-C were confirmed by western blot analysis and fluorescence microscopy to stably express the corresponding proteins. No differences were observed in the in vitro growth rates between any of these cells. However, co-culture of MSTO-A, MSTO-E and MSTO-F showed significant suppression of human umbilical endothelial cell growth in vitro compared with that of MSTO-C. Furthermore, a combination of any two among MSTO-A, MSTO-E and MSTO-F significantly enhanced efficacy. These results suggest that combinatorial anti-angiogenic gene therapy targeting different pathways of endothelial growth factor signaling has the potential for greater therapeutic efficacy than that of a single-agent regimen.
authors.affiliation: Department of Genetics, Hyogo College of Medicine, Nishinomiya, Hyogo, Japan.
authors.forename: Shuji
authors.initials: S
authors.lastname: Kubo
authors.affiliation: Department of Genetics, Hyogo College of Medicine, Nishinomiya, Hyogo, Japan.
authors.forename: Misato
authors.initials: M
authors.lastname: Takagi-Kimura
authors.affiliation: Departments of Cell Biology and Pathology, University of Miami, Miami, FL, USA.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Kasahara
mesh_terms: D020533:Angiogenesis Inhibitors / Q000235:genetics*; D043168:Angiostatins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D043169:Endostatins / Q000235:genetics / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008654:Mesothelioma / Q000235:genetics / Q000628:therapy*; D051379:Mice; D051497:Receptor, Fibroblast Growth Factor, Type 2 / Q000235:genetics / Q000378:metabolism
chemical_list: D020533:Angiogenesis Inhibitors; D043169:Endostatins; D043168:Angiostatins; C496353:FGFR2 protein, human; D051497:Receptor, Fibroblast Growth Factor, Type 2