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Dynamic changes in the characteristics of cationic lipidic vectors after exposure to mouse serum: implications for intravenous lipofection

Gene Therapy volume 6, pages 585–594 (1999)Cite this article

Abstract

Intravenous gene delivery via cationic lipidic vectors gives systemic gene expression particularly in the lung. In order to understand the mechanism of intravenous lipofection, a systematic study was performed to investigate the interactions of lipidic vectors with mouse serum emphasizing how serum affects the biophysical and biological properties of vectors of different lipid compositions. Results from this study showed that lipidic vectors underwent dynamic changes in their characteristics after exposure to serum. Addition of lipidic vectors into serum resulted in an immediate aggregation of vectors. Prolonged incubation of lipidic vectors with serum led to vector disintegration as shown in turbidity study, sucrose-gradient centrifugation analysis and fluorescence resonance energy transfer (FRET) study. Vector disintegration was associated with DNA release and degradation as shown in EtBr intercalation assay and DNA digestion study. Serum-induced disintegration of vectors is a general phenomenon for all cationic lipidic vectors tested in this study. Yet, vectors of different lipid compositions vary greatly in the rate of disintegration. There is an inverse correlation between the disintegration rate of lipidic vectors and their in vivo transfection efficiency. Vectors with a rapid rate of disintegration such as those containing dioleoyl- phosphatidylethanolamine (DOPE) poorly stayed in the lung and were barely active in transfecting cells. In contrast, cholesterol-containing vectors that had a rapid aggregation and a slow disintegration were highly efficient in transfecting cells in vivo. The results of this study explain why cationic lipidic vectors of different lipid compositions have a dramatic difference in their in vivo transfection efficiency. These results also suggest that the study of the interactions of lipidic vectors with serum may serve as a predictive model for the in vivo efficiency of a lipidic vector. Further study of the numerous interactions of lipidic vectors with serum might lead to the development of a vector which can deliver a gene to target cells in a tissue-specific manner.

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References

  1. Ross G et al. Gene therapy in the United States: a five-year status report Hum Gene Ther 1996 7: 1781–1790

    Article  CAS  Google Scholar 

  2. Caplen NJ et al. Liposome-mediated CFTR gene transfer to the nasal epithelium of patients with cystic fibrosis Nature Med 1995 1: 39–46

    Article  CAS  Google Scholar 

  3. McLachlan G et al. Laboratory and clinical studies in support of cystic fibrosis gene therapy using pCMV-CFTR-DOTAP Gene Therapy 1996 3: 1113–1123

    CAS  PubMed  Google Scholar 

  4. Lee ER et al. Detailed analysis of structures and formulations of cationic lipids for efficient gene transfer to the lung Hum Gene Ther 1996 7: 1701–1717

    Article  CAS  Google Scholar 

  5. Stephan DJ et al. A new cationic liposome DNA complex enhances the efficiency of arterial gene transfer in vivo Hum Gene Ther 1996 7: 1803–1812

    Article  CAS  Google Scholar 

  6. Wheeler CJ et al. A novel cationic lipid greatly enhances plasmid DNA delivery and expression in mouse lung Proc Natl Acad Sci USA 1996 93: 11454–11459

    Article  CAS  Google Scholar 

  7. Zabner J et al. Cellular and molecular barriers to gene transfer by a cationic lipid J Biol Chem 1995 270: 18997–19007

    Article  CAS  Google Scholar 

  8. Dean DA . Import of plasmid DNA into the nucleus is sequence specific Exp Cell Res 1997 230: 293–302

    Article  CAS  Google Scholar 

  9. Sebestyen MG et al. DNA vector chemistry: the covalent attachment of signal peptides to plasmid DNA Nature Biotechnol 1998 16: 80–85

    Article  CAS  Google Scholar 

  10. Felgner PL et al. Nomenclature for synthetic gene delivery systems Hum Gene Ther 1997 8: 511–512

    Article  CAS  Google Scholar 

  11. Zhu N, Liggitt D, Liu Y, Debs R . Systemic gene expression after intravenous DNA delivery into adult mice Science 1993 261: 209–211

    Article  CAS  Google Scholar 

  12. Liu Y et al. Factors influencing the efficiency of cationic liposome-mediated intravenous gene delivery Nature Biotechnol 1997 15: 167–173

    Article  CAS  Google Scholar 

  13. Hong KL, Zheng WW, Baker A, Papahadjopoulos D . Stabilization of cationic liposome-plasmid DNA complexes by polyamines and poly(ethylene glycol)-phospholipid conjugates for efficient in vivo gene delivery FEBS Lett 1997 400: 233–237

    Article  CAS  Google Scholar 

  14. Templeton NS et al. Improved DNA:liposome complexes for increased systemic delivery and gene expression Nature Biotechnol 1997 15: 647–652

    Article  CAS  Google Scholar 

  15. Liu F, Qi H, Huang L, Liu D . Factors controlling the efficiency of cationic lipid-mediated transfection in vivo via intravenous administration Gene Therapy 1997 4: 517–523

    Article  CAS  Google Scholar 

  16. Li S, Huang L . In vivo gene transfer via intravenous administration of cationic lipid-protamine-DNA (LPD) complexes Gene Therapy 1997 4: 891–900

    Article  CAS  Google Scholar 

  17. Hofland HEJ et al. In vivo gene transfer by intravenous administration of stable cationic lipid/DNA complex Pharm Res 1997 14: 742–749

    Article  CAS  Google Scholar 

  18. Huang L, Li S . Liposomal gene delivery: a complex package Nature Biotechnol 1997 15: 620–621

    Article  CAS  Google Scholar 

  19. Barron LG, Meyer KB, Szoka FC . Effects of complement depletion on the pharmacokinetics and gene delivery mediated by cationic lipid-DNA complexes Hum Gene Ther 1998 9: 315–323

    Article  CAS  Google Scholar 

  20. Li S, Rizzo MA, Bhattacharya S, Huang L . Characterization of cationic lipid-protamine-DNA (LPD) complexes for intravenous gene delivery Gene Therapy 1998 5: 930–937

    Article  CAS  Google Scholar 

  21. Gershon H, Ghirlando R, Guttman SB, Minsky A . Mode of formation and structural features of DNA-cationic liposome complexes used for transfection Biochemistry 1993 32: 7143–7151

    Article  CAS  Google Scholar 

  22. Xu Y, Szoka FC . Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection Biochemistry 1996 35: 5616–5623

    Article  CAS  Google Scholar 

  23. Selvin PR . Fluorescence resonance energy transfer Meth Enzymol 1995 246: 301–335

    Google Scholar 

  24. McLean JW et al. Organ-specific endothelial cell uptake of cationic liposome–DNA complexes in mice Am J Physiol Heart Circ Physiol 1997 42: H387–H404

    Article  Google Scholar 

  25. Litzinger DC, Huang L . Phosphatidylethanolamine liposomes: drug delivery, gene transfer and immunodiagnostic applications Biochim Biophys Acta 1992 1113: 201–227

    Article  CAS  Google Scholar 

  26. Bonte F, Juliano RL . Interactions of liposomes with serum proteins Chem Phys Lipids 1986 40: 359–372

    Article  CAS  Google Scholar 

  27. Sambrook J, Fritsch EF, Maniatis T . Molecular Cloning, A Laboratory Manual Cold Spring Laboratory Harbor Press: Cold Spring Harbor, NY 1994

    Google Scholar 

  28. Prensky W . The radioiodination of RNA and DNA to high specific activities Meth Cell Biology 1976 13: 121–152

    Article  CAS  Google Scholar 

  29. Sixou S et al. Intracellular oligonucleotide hybridization detected by fluorescence resonance energy transfer (FRET) Nucleic Acids Res 1994 22: 662–668

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Pharmacology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    S Li, W-C Tseng, S-P Wu & L Huang

  2. Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    D Beer Stolz & S C Watkins

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  1. S Li
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  2. W-C Tseng
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Li, S., Tseng, WC., Stolz, D. et al. Dynamic changes in the characteristics of cationic lipidic vectors after exposure to mouse serum: implications for intravenous lipofection. Gene Ther 6, 585–594 (1999). https://doi.org/10.1038/sj.gt.3300865

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