title: RNAi-mediated gene silencing in tumour tissue using replication-competent retroviral vectors.
abstract: RNAi represents a powerful technology to specifically downregulate the expression of target genes. For cancer research and therapy, an efficient in vivo delivery system is supposed to distribute RNAi to all tumour cells upon systemic administration. We present replication-competent murine leukaemia virus (MLV) vectors, which deliver RNAi to tumour tissue upon tail vein injection. In HT1080 cells stably expressing GFP or luciferase, GFP expression was suppressed by more than 80% and luciferase (luc) activity by more than 90%, even when only 0.1% of the cells were initially infected with reporter gene specific vectors. To demonstrate its potential, PLK1- and MMP14-specific small hairpin RNA expression cassettes were applied in the system. Upon infection, PLK1 and MMP14 levels were reduced on mRNA and protein level. MLV-shPLK1-infected cells were arrested in the G2-phase and underwent apoptosis. MLV-shMMP14-infected cells showed reduced MMP2 activity, as well as substantially reduced invasion and tumour growth. In vivo, MLV-shLuc silenced luc expression in HT1080-luc tumour tissue by more than 80% and MLV-shPLK1 reduced tumour growth substantially, demonstrating the therapeutic relevance of this system. This RNAi vector system allows long-term downregulation of target gene expression as well as efficient delivery to and distribution throughout tumour tissue in vivo.
authors.affiliation: Division of Medical Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Schaser
authors.forename: C
authors.initials: C
authors.lastname: Wrede
authors.forename: L
authors.initials: L
authors.lastname: Duerner
authors.forename: K
authors.initials: K
authors.lastname: Sliva
authors.forename: K
authors.initials: K
authors.lastname: Cichutek
authors.forename: B
authors.initials: B
authors.lastname: Schnierle
authors.forename: C J
authors.initials: CJ
authors.lastname: Buchholz
mesh_terms: D000818:Animals; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000235:genetics; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D059565:G2 Phase Cell Cycle Checkpoints / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics* / Q000627:therapeutic use*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics*; D008156:Luciferases / Q000378:metabolism; D053511:Matrix Metalloproteinase 14 / Q000235:genetics; D051379:Mice; D016513:Mice, SCID; D009369:Neoplasms / Q000378:metabolism* / Q000628:therapy*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D000097869:Polo-Like Kinase 1
chemical_list: D018797:Cell Cycle Proteins; D017931:DNA Primers; D049452:Green Fluorescent Proteins; D008156:Luciferases; D053511:Matrix Metalloproteinase 14; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D000097869:Polo-Like Kinase 1