title: Application of poly(2-(dimethylamino)ethyl methacrylate)-based polyplexes for gene transfer into human ovarian carcinoma cells.
abstract: Previously, attempts were made in our laboratory to transfect human ovarian cancer (OVCAR-3) cells, growing in the peritoneal cavity of nude mice, by intraperitoneal administration of poly(2-(dimethylamino)ethyl methacrylate) (pDMAEMA)-based polyplexes. However, hardly any transfection of the OVCAR-3 cells was observed. The aim of the present study was to examine whether pDMAEMA-polyplexes can transfect OVCAR-3 cells in vivo at DNA doses much higher than used previously [J. Gene Med. 1 (1999) 156-158]. We also explored a specific targeting strategy based on the use of folic acid (FA) as a targeting ligand directed against the folate receptor overexpressed on OVCAR-3 cells. Luciferase expression by OVCAR-3 cells mediated by pDMAEMA-based polyplexes was evaluated in the mouse i.p. OVCAR-3 xenograft model of ovarian cancer. By virtue of new formulation options, we were able to administer polyplex dispersions into OVCAR-3 bearing mice at much larger doses (75-120 microg DNA) than used previously (15 microg). The feasibility of folate-mediated targeting of the polyplexes was studied after coupling of FA to preformed polyplexes with poly(ethylene glycol) (PEG) as a spacer. Intraperitoneal administration of naked pLuc plasmid did not result in significant gene expression by the tumor cells. Administration of uncoated, positively charged pDMAEMA-based polyplexes at a DNA dose of 75-120 microg yielded significant transfection activity. However, also considerable gene expression was observed in non-target cells. To avoid transfection of non-target cells, an active targeting strategy based on the use of FA was studied. At a dose of 75 microg DNA (N/P 5), the folate-targeting approach yielded about 10-fold lower luciferase transfection levels in organs lined by the mesenthelial layer. This beneficial site-avoidance effect was achieved without compromising the degree of tumor cell transfection. Successful transfection of OVCAR-3 cells growing in the peritoneal cavity of nude mice can be achieved by i.p. administration of polyplexes at doses between 75 and 120 microg DNA. It was further demonstrated that active targeting of polyplexes to OVCAR-3 cells growing in the peritoneal cavity of mice is a realistic possibility to avoid transfection of non-target cells.
authors.affiliation: Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, P.O. Box 80082, 3508 TB, Utrecht, The Netherlands.
authors.forename: Ferry J
authors.initials: FJ
authors.lastname: Verbaan
authors.forename: Peter
authors.initials: P
authors.lastname: Klein Klouwenberg
authors.forename: Jan Hein
authors.initials: JH
authors.lastname: van Steenis
authors.forename: Cor J
authors.initials: CJ
authors.lastname: Snel
authors.forename: Otto
authors.initials: O
authors.lastname: Boerman
authors.forename: Wim E
authors.initials: WE
authors.lastname: Hennink
authors.forename: Gert
authors.initials: G
authors.lastname: Storm
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D005260:Female; D005492:Folic Acid / Q000008:administration & dosage; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007274:Injections, Intraperitoneal; D008156:Luciferases / Q000235:genetics; D008689:Methacrylates / Q000737:chemistry*; D051379:Mice; D009757:Nylons / Q000737:chemistry*; D010051:Ovarian Neoplasms / Q000473:pathology / Q000628:therapy*; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D014162:Transfection; D023041:Xenograft Model Antitumor Assays
chemical_list: D008689:Methacrylates; D009757:Nylons; C407037:poly(2-(dimethylamino)ethyl methacrylate); D004247:DNA; D005492:Folic Acid; D008156:Luciferases