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Plasmid Delivery in the Rat Brain

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Bioscience Reports

Abstract

Neurodegenerative diseases as a class do not have effective pharmacotherapies. This is due in part to a poor understanding of the pathologies of the disease processes, and the lack of effective medications. Gene delivery is an attractive possibility for treating these diseases. For the paradigm to be effective, efficient, safe and versatile vectors are required. In this study we evaluated three plasmid delivery systems for transgene expression in the rat hippocampus. Two of these systems were designed to have enhanced intracellular biodegradability. It was hypothesized that this system would be less toxic and could increase the free (non-vector) associated plasmids within the cell, leading to increased transgene activity. Polyethylenimine (PEI) and r-AAV-2 (recombinant adeno associated virus-2) were used as positive, non-viral and viral controls respectively, in the in vivo experiments. The results from the studies indicate there is a distinct difference between the various vectors in terms of total cells transfected, type of cell transfected, and toxicity. Non-viral systems were effective at transfecting both neurons and glia cells within the hippocampus, while the r-AAV-2 transfected mainly neurons. In summary, plasmid-mediated systems are effective for transgene expression within the brain and deserve further study.

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REFERENCES

  1. Kofler, P., Weisenhofer, B., Rehrl, C., Baier, G., Stockhammer, G., and Humpel, C. (1998) Liposome-mediated gene transfer into established CNS cell lines, primary glial cells, and in vivo. Cell Transplant 7(2):175-185.

    Google Scholar 

  2. Griffit, W., Glick, R. P., Lichtor, T., and Cohen, E. P. (1998) Survival and toxicity of an allogeneic cytokine-secreting fibroblast vaccine in the central nervous system. Neurosurgery 42(2):335-340.

    Google Scholar 

  3. Rainov, N. G., Koch, S., Sena-Esteves, M., and Berens, M. E. (2000) Characterization of a canine glioma cell line as related to established experimental brain tumor models. J. Neuropathol. Exp. Neurol. 59(7):607-613.

    Google Scholar 

  4. Princen, F., Lechanteur, C., Lopez, M., Gielen, J., Bours, V., and Merville, M. P. (2000) Similar efficiency of DNA-liposome complexes and retrovirus-producing cells for HSV-tk suicide gene therapy of peritoneal carcinomatosis. J. Drug Target 8(2):79-89.

    Google Scholar 

  5. Shi, N. and Pardridge, W. M. Noninvasive gene targeting to the brain. Proc. Natl. Acad. Sci. USA 97(13):7567-7572.

  6. Li, S. et al. (2001) Intramuscular electroporation delivery of IFN-alpha gene therapy for inhibition of tumor growth located at a distant site. Gene Ther. 8(5):400-407.

    Google Scholar 

  7. Auricchio, A., Zhou, R., Wilson, J. M., and Glickson, J. D. (2001) In vivo detection of gene expression in liver by 31P nuclear magnetic resonance spectroscopy employing creatine kinase as a marker gene. Proc. Natl. Acad. Sci. USA 98(9):5205-5210.

    Google Scholar 

  8. Chen, Z. Y., Yant, S. R., He, C. Y., Meuse, L., Shen, S., and Kay, M. A. (2001) Linear DNAs concatermerize in vivo and result in sustained transgene expression in mouse liver. Mol. Ther. 3(3):403-410.

    Google Scholar 

  9. Tsujie, M., Isaka, Y., Nakamura, H., Kaneda, Y., Imai, E., and Hori, M. (2001) Prolonged transgene expression in glomeruli using an EBV replicon vector system combined with HVJ liposomes. Kidney Int. 59(4):1390-1396.

    Google Scholar 

  10. Agarwal, M., Austin, T. W., Morel, F., Chen, J., Bohnlein, E., and Plavec, I. Scaffold attachment region-mediated enhancement of retroviral vector expression in primary T cells. J. Virol. 72(5):3720-3728.

  11. Philip, R. et al. (1994) Efficient and sustained gene expression in primary T lymphocytes and primary and cultured tumor cells mediated by adeno-associated virus plasmid DNA complexed to cationic liposomes. Mol. Cell Biol. 14(4):2411-2418.

    Google Scholar 

  12. Zhou, H. et al. (1995) Enhanced adeno-associated virus vector expression by adenovirus proteincationic liposome complex. A novel and high efficient way to introduce foreign DNA into endothelial cells. Clin. Med. J. (Engl.) 108(5):332-337.

    Google Scholar 

  13. Houk, B., Hochhaus, G., and Hughes, J. A. (1999) In vitro kinetics of plasmid DNA. Pharm. Sci. in press (Volume 3).

  14. Levy, M. Y., Barron, L. G., Meyer, K. B., and Szoka, F. C. Jr. (1996) Characterization of plasmid DNA transfer into mouse skeletal muscle: evaluation of uptake mechanism, expression and secretion of gene products into blood. Gene Ther. 3(3):201-211.

    Google Scholar 

  15. Ruan, H., Su, H., Hu, L., Lamborn, K. R., Kan, Y. W., and Deen, D. F. (2001) A hypoxia-regulated adeno-associated virus vector for cancer-specific gene therapy. Neoplasia 3(3):255-263.

    Google Scholar 

  16. Nguyen, J. B., Sanchez-Pernaute, R., Cunningham, J., and Bankiewicz, K. S. (2001) Convectionenhanced delivery of r-AAV-2 combined with heparin increases TK gene transfer in the rat brain. Neuroreport 12(9):1961-1964.

    Google Scholar 

  17. Felgner, J. H. et al. (1994) Enhanced gene delivery and mechanisms studies with a novel series of cationic lipid formulations. Journ. Biol. Chem. 269(4):2550-2561.

    Google Scholar 

  18. Dan, N. (1998) The structure of DNA complexes with cationic liposomes-cylindrical or flat bilayers. Biochimica et Biophysica Acta 1369:34-38.

    Google Scholar 

  19. Papadopoulos, M. C., Giffard, R. G., and Bell, B. A. (2000) Principles of gene therapy: potential applications in the treatment of cerebral ischaemia. Br. J. Neursurg. 14(5):407-414.

    Google Scholar 

  20. Wiesenhofer, B., Weis, C., and Humpel, C., Glial cell line-derived neurotrophic factor (GDNF) is a proliferation factor for rat C6 glioma cells: evidence from antisense experiments. Antisense Nucleic Acid Drug Dev. 10(5):311-321.

  21. Zou, L. L. et al. (1999) Liposome-mediated NGF gene transfection following neuronal injury: potential therapeutic applications. Gene Ther. 6(6):994-1005.

    Google Scholar 

  22. Behr, J.-P., Demeneix, B, Loeffler, J.-P., and Perez-Mutul, J. (1989) Efficient gene transfer into mammalian primary endocrine cells with lipopolyamine-coated DNA. Proc. Natl. Acad. Sci. USA 86:6982-6986.

    Google Scholar 

  23. Tang, F. and Hughes, J. A. (1998) Introduction of a disulfide bond into a cationic lipid enhances transgene expression of plasmid DNA. Biochemical and Biophysical Research Communications 242:141-145.

    Google Scholar 

  24. Tang, F. and Hughes, J. A. (1999) Use of dithiodiglycolic acid as a tether for cationic lipids decreases the cytotoxicity and increases transgene expression of plasmid DNA in vitro. Bioconjug. Chem. 10(5):791-796.

    Google Scholar 

  25. Ajmani, P. S., Tang, F., Krishnaswami, S., Meyer, E. M., Sumners, C., and Hughes, J. A. (1999) Enhanced transgene expression in rat brain cell cultures with a disulfide containing cationic lipid. Neuroscience Letters 277:141-144.

    Google Scholar 

  26. Tang, F. and Hughes, J. A. (1999) Synthesis of a single-tailed cationic lipid and investigation of its transfection. J. Controlled Release 62(3):345-358.

    Google Scholar 

  27. Klein, R. L., Hirko, A. C., Meyers, C. A., Grimes, J. R., Muzyczka, N., and Meyer, E. M. (2000) NGF gene transfer to intrinsic basal forebrain neurons increases cholinergic cell size and protects from age-related, spatial memory deficits in middle-aged rats. Brain Res. 875: (1-2):144-151.

    Google Scholar 

  28. Tang, F. and Hughes, J. A. (2001) Cationic lipidic vectors for plasmid DNA delivery. Journal of Pharmaceutical Sciences. In press.

  29. Korbo, L., Ladefoged, O., Lam, H. R., Ostergaard, G., West, M. J. and Arlien-Soborg, P. (1996) Neuronal loss in hippocampus in rats exposed to toluene. Neurotoxicology 17(2):359-366.

    Google Scholar 

  30. West, M. J. and Slomianka, L. (1998) Total number of neurons in the layers of the human entorhinal cortex. Hippocampus 8(1):69-82.

    Google Scholar 

  31. West, M. J., Slomianka, L., and Gundersen, H. J. (1991) Unbiased stereological estimation of the total number of neurons in the subdivisions of the rat hippocampus using the optical fractionator. Anat Rec. 231(4):482-497.

    Google Scholar 

  32. Siegel, G. J. and Chauhan, N. B. (2000) Neurotrophic factors in Alzheimer's and Parkinson's diseased brain. Brain Res. Rev. 33(2-3):199-227.

    Google Scholar 

  33. Apfel, S. C. (2001) Neurotrophic factor therapy-prospects and problems. Clin. Chem. Lab. Med. 39(4):351-355.

    Google Scholar 

  34. Bartlett, J. S., Samulski, R. J., and McCown, T. J. (1998) Selective and rapid uptake of adenoassociated virus type 2 in brain. Hum. Gene Ther. 9(8):1181-1186.

    Google Scholar 

  35. Klein, R. L., Meyer, E. M., Peel, A. L., Zolotukhin, S., Muzyczka, N., Meyers, C., and King, M. A. (1998) Neuron-specific transduction in rat septohippocampal or nigrostriatal pathway by recombinant adeno-associated virus vectors. Exp. Neurol. 150:183-194.

    Google Scholar 

  36. Abdallah, B., Sachs, L., and Demeneix, B. A. (1995) Non-viral gene transfer: applications in developmental biology and gene therapy. Biol. Cell 85(1):1-7.

    Google Scholar 

  37. Abdallah, B., Hassan, A., Benoist, C., Goula, D., Behr, J. P., and Demeneix, B. A. (1996) A powerful nonviral vector for in vivo gene transfer into the adult mammalian brain: polyethylenimine. Hum. Gene Ther. 7(16):1947-1954.

    Google Scholar 

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Authors and Affiliations

  1. Department of Pharmaceutics, University of Florida, College of Pharmacy, JHMHC Box 100494, Gainesville, FL, 32610

    A.C. Hirko, D.D. Buethe & J.A. Hughes

  2. Department of Pharmacology, University of Florida, College of Pharmacy, JHMHC Box 100494, Gainesville, FL, 32610

    E.M. Meyer

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  1. A.C. Hirko
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  2. D.D. Buethe
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  3. E.M. Meyer
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  4. J.A. Hughes
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Correspondence to J.A. Hughes.

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Hirko, A., Buethe, D., Meyer, E. et al. Plasmid Delivery in the Rat Brain. Biosci Rep 22, 297–308 (2002). https://doi.org/10.1023/A:1020194807665

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