Evaluation of a new biocompatible poly(N-(morpholino ethyl methacrylate)-based copolymer for the delivery of ruthenium oligonucleotides, targeting HPV16 E6 oncogene
[en] 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 RuII 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®.
Research center :
Centre Interfacultaire de Recherche du Médicament - CIRM
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Reschner, Anca ; Université de Liège - ULiège > Immunologie et vaccinologie
Shim, Yong Ho
Dubois, Philippe
Delvenne, Philippe ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Evrard, Brigitte ; Université de Liège - ULiège > Département de pharmacie > Pharmacie galénique
Marcelis, Lionel
Moucheron, Cécile
Kirsch-De Mesmaeker, Andrée
Defrancq, Eric
Raes, Martine
Piette, Jacques ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Virologie - Immunologie
Collard, Laurence ; Université de Liège - ULiège > Département de pharmacie > Pharmacie galénique
Piel, Géraldine ; Université de Liège - ULiège > Département de pharmacie > Pharmacie galénique
Evaluation of a new biocompatible poly(N-(morpholino ethyl methacrylate)-based copolymer for the delivery of ruthenium oligonucleotides, targeting HPV16 E6 oncogene
M. Mizuuchi, Y. Hirohashi, T. Torigoe, T. Kuroda, K. Yasuda, Y. Shimizu, T. Saito, and N. Sato, Novel oligomannose liposome-DNA complex DNA vaccination efficiently evokes anti-HPV E6 and E7 CTL responses. Experimental and Molecular Pathology 92, 185 (2012).
T. Dutta, M. Burgess, N. A. McMillan, and H. S. Parekh, Dendrosome-based delivery of siRNA against E6 and E7 oncogenes in cervical cancer. Nanotechnology Biology and Medicine 6, 463 (2010).
S. Lai, E. O'Hanlon, S. Harrold, S. T. Man, Y. Y. Wang, R. Cone, and J. Hanes, Rapid transport of large polymeric nanoparticles in fresh undiluted human mucus. PNAS 104, 1482 (2007).
R. Ghittoni, R. Accardi, U. Hasan, T. Gheit, B. Sylla, and M. Tommasino, The biological properties of E6 and E7 oncoproteins from human papillomaviruses. Virus Genes 40, 1 (2010).
A. L. Jonson, L. M. Rogers, S. Ramakrishnan, and L. S. Downs, Gene silencing with siRNA targeting E6/E7 as a therapeutic intervention in a mouse model of cervical cancer. Gynecologic Oncology 111, 356 (2008).
L. M. Alvarez-Salas and J. A. DiPaolo, Molecular Approaches to Cervical Cancer Therapy. Curr. Drug Discov. Technol. 4, 208 (2007).
K. A. Woodrow, Y. Cu, C. J. Booth, J. K. Saucier-Sawyer, M. J. Wood, and W. M. Saltzman, Intravaginal gene silencing using biodegradable polymer nanoparticles densely loaded with small-interfering RNA. Nat. Mater. 8, 526 (2009).
M. Morille, C. Passirani, A. Vonarbourg, A. Clavreul, and J. P. Benoit, Progress in developing cationic vectors for non-viral systemic gene therapy against cancer. Biomaterials 29, 3477 (2008).
M. A. Mintzer and E. E. Simanek, Nonviral vectors for gene delivery. Chem. Rev. 109, 259 (2009).
J. C. Tseng, T. Granot, V. Digiacomo, B. Levin, and D. Meruelo, Enhanced specific delivery and targeting of oncolytic Sindbis viral vectors by modulating vascular leakiness in tumor. Cancer Gene Ther. 17, 244 (2010).
E. E. Thacker, M. Nakayama, B. F. Smith, R. C. Bird, Z. Muminova, and T. V. Strong, A genetically engineered adenovirus vector targeted to CD40 mediates transduction of canine dendritic cells and promotes antigen-specific immune responses in vivo. Vaccine 23, 7116 (2009).
M. Huang, Z. Chen, S. Hu, F. Jia, Z. Li, G. Hoyt, R. C. Robbins, M. A. Kay, and J. C. Wu, Novel minicircle vector for gene therapy in murine myocardial infarction. Circulation 15, S230-7 (2009).
C. E. Thomas, A. Ehrhardt, and M. A. Kay, Progress and problems with the use of viral vectors for gene therapy. Nat. Rev. Genet. 4, 346 (2003).
A. Schallon, V. Jérôme, A. Walther, C. V. Synatschke, A. Müller, and R. Freitag, Performance of three PDMAEMA-based polycation architectures as gene delivery agents in comparison to linear and branched PEI. Reactive and Functional Polymers 70, 1 (2010).
P. van de Wetering, J. Cherng, H. Talsma, D. J. Crommelin, and W. E. Hennink, 2-(dimethylamino)ethyl methacrylate based (co)polymers as gene transfer agents. J. Controlled Release 53, 145 (1998).
S. Pirotton, C. Muller, N. Pantoustier, F. Botteman, S. Collinet, C. Grandfils, G. Dandrifosse, P. Degee, P. Dubois, and M. Raes, Enhancement of transfection efficiency through rapid and noncovalent post-PEGylation of poly(dimethylaminoethyl methacrylate)/DNA complexes. Pharm. Res. 21, 1471 (2004).
C. Zhu, S. Jung, S. Luo, F. Meng, X. Zhu, T. G. Park, and Z. Zhong, Co-delivery of siRNA and paclitaxel into cancer cells by biodegradable cationic micelles based on PDMAEMA-PCL-PDMAEMA triblock copolymers. Biomaterials 31, 2408 (2010).
R. A. Jones, M. H. Poniris, and M. R. Wilson, PDMAEMA is internalised by endocytosis but does not physically disrupt endosomes. J. Controlled Release 96, 379 (2004).
N. Van Overstraeten-Schlögel, Y. H. Shim, V. Tevel, G. Piel, J. Piette, P. Dubois, and M. Raes, Assessment of new biocompatible Poly(N-(morpholino)ethyl methacrylate)-based copolymers by transfection of immortalized keratinocytes. Drug Delivery 16, 102 (2012).
S. Le Gac, S. Rickling, P. Gerbaux, E. Defrancq, C. Moucheron, and A. Kirsch-De Mesmaeker, A photoreactive ruthenium (II) complex tethered to a guanine-containing oligonucleotide: A biomolecular tool that behaves as a "Seppuku Molecule," Angew. Chem. Int. Ed. 48, 1122 (2009).
A. Reschner, S. Bontems, S. Le Gac, J. Lambermont, L. Marcélis, E. Defrancq, P. Hubert, C. Moucheron, A. Kirsch-De Mesmaeker, M. Raes, J. Piette, and P. Delvenne, Ruthenium oligonucleotides, targeting HPV16 E6 oncogene, inhibit the growth of cervical cancer cells under illumination by a mechanism involving p53. Gene Therapy advance online publication, doi:10.1038/gt.2012.54 (2012).
Y. H. Shim, F. Bougard, O. Coulembier, R. Lazzaroni, and P. Dubois, Synthesis and characterization of original 2-(dimethylamino)ethyl methacrylate/poly(ethyleneglycol) star-copolymers. European Polymer J. 44, 3715 (2008).
S. Deroo, E. Defrancq, C. Moucheron, A. Kirsch-De Mesmaeker, and P. Dumy, Synthesis of an oxyamino-containing phenanthroline derivate for the efficient preparation of phenanthroline oligonucleotide oxime conjugates. Tetrahedron Lett. 44, 8379 (2003).
M. Villien, S. Deroo, E. Gicquel, E. Defrancq, C. Moucheron, A. Kirsch-De Mesmaeker, and P. Dumy, The oxime bound formation as an efficient tool for the conjugation of Ru complexes to oligonucleotides and peptides. Tetrahedron 63, 11299 (2007).
J. Y. Cherng, H. Talsma, R. Verrijk, D. J. Crommelin, and W. E. Hennink, The effect of formulation parameters on the size of poly((2-dimethylamino)ethyl methacrylate)-plasmid complexes. Journal of Pharmaceutics and Biopharmaceutics 47, 215 (1999).
M. Yoshinouchi, T. Yamada, M. Kizaki, J. Fen, T. Koseki, Y. Ikeda, T. Nishihara, and K. Yamamoto, In vitro and in vivo growth suppression of human papillomavirus 16-positive cervical cancer cells by E6 siRNA. Mol. Ther. 8, 762 (2003).
M. Jiang and J. Milner, Selective silencing of viral gene expression in HPV-positive human cervical carcinoma cells treated with siRNA, a primer of RNA interference. Oncogene 21, 6041 (2002).
