A Novel Non-Viral Vector for DNA Delivery Based on Low Molecular Weight, Branched Polyethylenimine: Effect of Molecular Weight on Transfection Efficiency and Cytotoxicity

Pharmaceutical Research - Tập 16 Số 8 - Trang 1273-1279 - 1999
Fischer, Dagmar1, Bieber, Thorsten2, Li, Youxin3, Elsässer, Hans-Peter2, Kissel, Thomas1
1Department of Pharmaceutics and Biopharmacy, University of Marburg, Philipps-University, Marburg, Germany
2Department of Cell Biology, University of Marburg, Marburg, Germany
3Schwarz Pharma AG, Monheim, Germany

Tóm tắt

Purpose. Low molecular weight branched polyethylenimine (LMW-PEI) was synthesized and studied as a DNA carrier for gene delivery with regard to physico-chemical properties, cytotoxicity, and transfection efficiency. Methods. The architecture of LMW-PEI, synthesized by acid catalyzed ring-opening polymerization of aziridine was characterized by size exclusion chromatography in combination with laser light scattering and 13C-NMR-spectroscopy. In vitro cytotoxic effects were quantified by LDH and MTT assay and visualized by transmission electron microscopy. The potential for transgene expression was monitored in ECV304 cells using luciferase driven by a SV40 promoter as reporter gene system. Results. LMW-PEI (Mw 11′900 D) with a low degree of branching was synthesized as a DNA carrier for gene delivery. In contrast to high molecular weight polyethylenimines (HMW-PEI; Mw l′616′OOO D), the polymer described here showed a different degree of branching and was less cytotoxic in a broad range of concentrations. As demonstrated by transmission electron microscopy the LMW-PEI formed only small aggregates which were efficiently taken up by different cells in the presence of serum, most likely by an endocytic pathway. LMW-PEI yielded transfection efficiencies measured via expression of the reporter gene luciferase which were up to two orders of magnitude higher than those obtained with HMW-PEI. The reporter gene expression was concentration dependent, but in contrast to lipofection independent of serum addition. Conclusions. The LMW-PEI described here is a new, highly efficient, and non-cytotoxic vector with a favorable efficiency/toxicity profile for gene therapeutic applications.

Từ khóa


Tài liệu tham khảo

citation_journal_title=Hum. Gene Ther.; citation_title=Nonviral gene therapy: the promise of genes as pharmaceutical products; citation_author=F. D. Ledley; citation_volume=6; citation_publication_date=1995; citation_pages=1129-1144; citation_id=CR1

citation_journal_title=Science; citation_title=The basic science of gene therapy; citation_author=R. C. Mulligan; citation_volume=260; citation_publication_date=1993; citation_pages=926-932; citation_id=CR2

citation_journal_title=Pharm. Acta Helv.; citation_title=The new frontier: gene and oligonucleotide therapy; citation_author=H. Schreier; citation_volume=68; citation_publication_date=1994; citation_pages=145-159; citation_id=CR3

citation_journal_title=J. Contr. Rel.; citation_title=Controllable gene therapy: pharmaceutics of non-viral gene delivery systems; citation_author=E. Tomlinson, A. Rolland; citation_volume=39; citation_publication_date=1996; citation_pages=357-371; citation_id=CR4

citation_journal_title=Chimia; citation_title=The proton sponge: a trick to enter cells the viruses did not exploit; citation_author=J. P. Behr; citation_volume=51; citation_publication_date=1997; citation_pages=34-36; citation_id=CR5

citation_journal_title=Adv. Drug Del. Rev.; citation_title=Gene transfer with lipospermines and polyethy-lenimines; citation_author=J. S. Remy, B. Abdallah, M. A. Zanta, O. Boussif, J. P. Behr, B. A. Demeneix; citation_volume=30; citation_publication_date=1998; citation_pages=85-95; citation_id=CR6

citation_journal_title=Proc. Natl. Acad. Sci. USA; citation_title=A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethy-lenimine; citation_author=O. Boussif, F. Lezoulac'h, M. A. Zanta, M. Mergny, D. Scherman, B. Demeneix, J. P. Behr; citation_volume=92; citation_publication_date=1995; citation_pages=7297-7303; citation_id=CR7

citation_journal_title=Nucl. Acid Res.; citation_title=Delivery of bacterial artificial chromosomes into mammalian cells with psoralen-inactivated adenovirus carrier; citation_author=A. Baker, M. Cotten; citation_volume=25; citation_publication_date=1997; citation_pages=1950-1956; citation_id=CR8

citation_journal_title=Gene Therapy; citation_title=Adenovirus enhancement of polyethylenimine-mediated transfer of regulated genes in differentiated cells; citation_author=C. Meunier-Durmont, H. Grimal, L. M. Sachs, B. A. Demeneix, C. Forest; citation_volume=4; citation_publication_date=1997; citation_pages=808-814; citation_id=CR9

citation_journal_title=Gene Ther.; citation_title=Polyethylenimine (PEI) is a simple, inexpensive and effective reagent for condensing and linking plasmid DNA to adenovirus for gene delivery; citation_author=A. Baker, M. Salik, H. Lehmann, I. Killisch, V. Mautner, G. Lamm, C. Christofori; citation_volume=4; citation_publication_date=1997; citation_pages=773-782; citation_id=CR10

citation_journal_title=Gene Ther.; citation_title=Coupling of cell-binding ligands to polyethylenimine for targeted gene delivery; citation_author=R. Kircheis, A. Kichler, G. Wallner, M. Kursa, M. Ogris, T. Felzmann, M. Buchberger, E. Wagner; citation_volume=4; citation_publication_date=1997; citation_pages=409-418; citation_id=CR11

citation_journal_title=Gene Ther.; citation_title=Optimized galenics improve in vitro gene transfer with cationic molecules up to 1000-fold; citation_author=O. Boussif, M. A. Zanta, J. P. Behr; citation_volume=3; citation_publication_date=1996; citation_pages=1074-1080; citation_id=CR12

citation_journal_title=Hum. Gene Ther.; citation_title=A powerful nonviral vector for in vivo gene transfer into the adult mammalian brain: polyethylenimine; citation_author=B. Abdallah, A. Hassan, C. Benoist, D. Goula, J. P. Behr, B. A. Demeneix; citation_volume=7; citation_publication_date=1997; citation_pages=1947-1954; citation_id=CR13

citation_journal_title=Hum. Gene Ther.; citation_title=Nonviral gene delivery to the rat kidney with polyethy-lenimine; citation_author=A. Boletta, A. Benigni, J. Lutz, G. Remuzzi, M. S. Soria, L. Monaco; citation_volume=8; citation_publication_date=1997; citation_pages=1243-1251; citation_id=CR14

citation_journal_title=Proceed. Int. Symp. Control. Rel. Bioact. Mater.; citation_title=Size matters: molecular weight affects the efficiency of poly(ethylenimine) as gene delivery vehicle; citation_author=W. T. Godbey, K. K. Wu, A. G. Mikos; citation_volume=25; citation_publication_date=1998; citation_pages=230-231; citation_id=CR15

citation_title=The normal-coordinate method for polymer chains in dilute solution; citation_inbook_title=Rheology; citation_publication_date=1960; citation_id=CR16; citation_author=B. H. Zimm; citation_publisher=Academic Press

citation_journal_title=J. Immunol. Methods; citation_title=Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assay; citation_author=T. Mosmann; citation_volume=65; citation_publication_date=1983; citation_pages=55-63; citation_id=CR17

citation_journal_title=J. Cell Biol.; citation_title=Formaldehyde-glutaraldehyde fixatives containing trinitro compounds; citation_author=S. Ito, M. J. Karnovsky; citation_volume=39; citation_publication_date=1968; citation_pages=168-169; citation_id=CR18

citation_journal_title=J. Biophys. Biochem. Cytol.; citation_title=Improvements in epoxin resin embedding methods; citation_author=J. H. Luft; citation_volume=9; citation_publication_date=1961; citation_pages=409-414; citation_id=CR19

citation_journal_title=J. Macromol. Sci.; citation_title=Characterization of polyethylenimine; citation_author=C. R. Dick, G. E. Ham; citation_volume=A4; citation_publication_date=1997; citation_pages=1301-1314; citation_id=CR20

citation_journal_title=Proc. Intern. Symp. Control. Rel. Bioact. Mater.; citation_title=Comparative in vitro cytotoxicity studies of polycations for gene therapy; citation_author=D. Fischer, R. Zange, T. Kissel; citation_volume=24; citation_publication_date=1997; citation_pages=527-528; citation_id=CR21

citation_journal_title=Eur. J. Cell. Biol.; citation_title=Polyethylenimine: synthesis and in vitro cytotoxicity of a low molecular weight polycation for gene transfer; citation_author=D. Fischer, T. Bieber, H. P. Elsässer, T. Kissel; citation_volume=75; citation_issue=suppl.48; citation_publication_date=1998; citation_pages=108; citation_id=CR22

citation_journal_title=J. Contr. Rel.; citation_title=In vitro cytotoxicity of macromolecules in different cell culture systems; citation_author=S. Choksakulnimitr, S. Masuda, H. Tokuda, Y. Takakura, M. Hashida; citation_volume=34; citation_publication_date=1995; citation_pages=233-241; citation_id=CR23

citation_journal_title=Nature; citation_title=A membrane effect of basic polymers dependent on molecular size; citation_author=H. J. P. Ryser; citation_volume=215; citation_publication_date=1967; citation_pages=934-936; citation_id=CR24

citation_journal_title=Gene Ther.; citation_title=ExGene 500 is an effective vector for gene delivery to lung epithelial cells in vitro and in vivo; citation_author=S. Ferrari, E. Moro, A. Pettenazzo, J. P. Behr, F. Zacchello, M. Scarpa; citation_volume=4; citation_publication_date=1997; citation_pages=1100-1106; citation_id=CR25

citation_journal_title=Gene Ther.; citation_title=Polyethylenimine-based intravenous delivery of transgenes to mouse lung; citation_author=D. Goulda, C. Benoist, S. Mantero, G. Merlo, G. Levi, B. A. Demeneix; citation_volume=5; citation_publication_date=1998; citation_pages=1291-1295; citation_id=CR26

citation_journal_title=Gene Ther.; citation_title=Size, diffusability and transfection performance of linear PEI/DNA complexes in the mouse central nervous system; citation_author=D. Goulda, J. S. Remy, J. S. Erbacher, M. Wasowicz, G. Levi, B. Abdallah, B. A. Demeneix; citation_volume=5; citation_publication_date=1998; citation_pages=712-717; citation_id=CR27

citation_journal_title=Biochim. Biophys. Acta; citation_title=Lipophilic polylysines mediate efficient DNA transfection in mammalian cells; citation_author=X. Zhou, A. L. Klibanov, L. Huang; citation_volume=1065; citation_publication_date=1991; citation_pages=8-14; citation_id=CR28

Thông tin
Thông tin xuất bản

Pharmaceutical Research

Tập 16 Số 8

1273-1279

Thông tin tác giả