Evaluation of a new biocompatible poly(N-(morpholino ethyl methacrylate)-based copolymer for the delivery of ruthenium oligonucleotides, targeting HPV16 E6 oncogene.

Abstract

This study investigates the use of a new biocompatible block copolymer poly(2-(dimethylamino)ethyl methacrylate-N-(morpholino)ethyl methacrylate (PDMAEMA-b-PMEMA) for the delivery of a particular antisense oligonucleotide targeting E6 gene from human papilloma virus. This antisense oligonucleotide was derivatized with a polyazaaromatic Ru(II) complex which, under visible illumination, is able to produce an irreversible crosslink with the complementary targeted sequence. The purpose of this study is to determine whether by the use of a suitable transfection agent, it is possible to increase the efficiency of the antisense oligonucleotide targeting E6 gene, named Ru-P-4. In a recent study, we showed that Oligofectamine transfected Ru-P-4 antisense oligonucleotide failed to inhibit efficiently the growth of cervical cancer cell line SiHa, contrarily to the Ru-P-6 antisense oligonucleotide, another sequence also targeting the E6 gene. The ability of PDMAEMA-b-PMEMA to form polyplexes with optimal physicochemical characteristics was investigated first. Then the ability of the PDMAEMA-b-PMEMA/Ru-P-4 antisense oligonucleotide polyplexes to transfect two keratinocyte cell lines (SiHa and HaCat) and the capacity of polyplexes to inhibit HPV16+ cervical cancer cell growth was evaluated. PDMAEMA-b-PMEMA base polyplexes at the optimal molar ratio of polymer nitrogen atoms to DNA phosphates (N/P), were able to deliver Ru-P-4 antisense oligonucleotide and to induce a higher growth inhibition in human cervical cancer SiHa cells, compared to other formulations based on Oligofectamine.

• Authors: Anca Reschner, Yong Ho Shim, Philippe Dubois, Philippe Delvenne, Brigitte Evrard, Lionel Marcélis, Cécile Moucheron, Andrée Kirsch-De Mesmaeker, Eric Defrancq, Martine Raes, Jacques Piette, Laurence Collard, Géraldine Piel
• Journal: Journal of biomedical nanotechnology (J Biomed Nanotechnol) Vol. 9 Issue 8 Pg. 1432-40 (Aug 2013) ISSN: 1550-7033 [Print] United States
• PMID: 23926811 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• E6 protein, Human papillomavirus type 16
• Methylmethacrylates
• Morpholinos
• Oligonucleotides
• Oligonucleotides, Antisense
• Oncogene Proteins, Viral
• Repressor Proteins
• Ruthenium
• poly(ethylmethacrylate)

Topics

• Cells, Cultured
• Drug Delivery Systems
• Female
• Genetic Therapy (instrumentation, methods)
• Human papillomavirus 16 (drug effects, genetics)
• Humans
• Keratinocytes (drug effects, virology)
• Materials Testing
• Methylmethacrylates (chemistry)
• Morpholinos (chemistry)
• Oligonucleotides (administration & dosage)
• Oligonucleotides, Antisense (administration & dosage)
• Oncogene Proteins, Viral (antagonists & inhibitors, genetics)
• RNA Interference
• Repressor Proteins (antagonists & inhibitors, genetics)
• Ruthenium (chemistry)
• Transfection (instrumentation, methods)