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Development of efficient plasmid DNA transfer into adult rat central nervous system using microbubble-enhanced ultrasound

Gene Therapy volume 11, pages 1532–1539 (2004)Cite this article

Abstract

Although gene therapy might become a promising approach for central nervous system diseases, the safety issue is a serious consideration in human gene therapy. To overcome this problem, we developed an efficient gene transfer method into the adult rat brain based on plasmid DNA using a microbubble-enhanced ultrasound method, since microbubble-enhanced ultrasound has shown promise for transfecting genes into other tissues such as blood vessels. Using the microbubble-enhanced ultrasound method, luciferase expression was increased approximately 10-fold as compared to injection of naked plasmid DNA alone. Interestingly, the site of gene expression was limited to the site of insonation with intracisternal injection, in contrast to previous studies using viruses. Expression of the reporter gene, Venus, was readily detected in the central nervous system. The transfected cells were mainly detected in meningeal cells with intracisternal injection, and in glial cells with intrastriatal injection. There was no obvious evidence of tissue damage by microbubble-enhanced ultrasound. Overall, the present study demonstrated the feasibility of efficient plasmid DNA transfer into the central nervous system, providing a new option for treating various diseases such as tumors.

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References

  1. Eck SL et al. Treatment of recurrent or progressive malignant glioma with a recombinant adenovirus expressing human interferon-beta (H5.010CMVhIFN- beta): a phase I trial. Hum Gene Ther 2001; 12: 97–113.

    Article  CAS  PubMed  Google Scholar 

  2. Trask TW et al. Phase I study of adenoviral delivery of the HSV-tk gene and ganciclovir administration in patients with current malignant brain tumors. Mol Ther 2000; 1: 195–203.

    Article  CAS  PubMed  Google Scholar 

  3. During MJ, Kaplitt MG, Stern MB, Eidelberg D . Subthalamic GAD gene transfer in Parkinson disease patients who are candidates for deep brain stimulation. Hum Gene Ther 2001; 12: 1589–1591.

    CAS  PubMed  Google Scholar 

  4. Janson C et al. Clinical protocol. Gene therapy of Canavan disease: AAV-2 vector for neurosurgical delivery of aspartoacylase gene (ASPA) to the human brain. Hum Gene Ther 2002; 13: 1391–1412.

    Article  CAS  PubMed  Google Scholar 

  5. Schnell MA et al. Activation of innate immunity in nonhuman primates following intraportal administration of adenoviral vectors. Mol Ther 2001; 3: 708–722.

    Article  CAS  PubMed  Google Scholar 

  6. Dewey RA et al. Chronic brain inflammation and persistent herpes simplex virus 1 thymidine kinase expression in survivors of syngeneic glioma treated by adenovirus-mediated gene therapy: implications for clinical trials. Nat Med 1999; 5: 1256–1263.

    Article  CAS  PubMed  Google Scholar 

  7. Hsich G, Sena-Esteves M, Breakefield XO . Critical issues in gene therapy for neurologic disease. Hum Gene Ther 2002; 13: 579–604.

    Article  CAS  PubMed  Google Scholar 

  8. Baumgartner I et al. Lower-extremity edema associated with gene transfer of naked DNA encoding vascular endothelial growth factor. Ann Intern Med 2000; 132: 880–884.

    Article  CAS  PubMed  Google Scholar 

  9. Rosengart TK et al. Angiogenesis gene therapy: phase I assessment of direct intramyocardial administration of an adenovirus vector expressing VEGF121 cDNA to individuals with clinically significant severe coronary artery disease. Circulation 1999; 100: 468–474.

    Article  CAS  PubMed  Google Scholar 

  10. Schwartz B et al. Gene transfer by naked DNA into adult mouse brain. Gene Therapy 1996; 3: 405–411.

    CAS  PubMed  Google Scholar 

  11. Sekiguchi K, Yasuzumi F, Morishita R . Exogenous expression of hepatocyte growth factor (HGF) in rat striatum by naked plasmid DNA. Neurosci Res 2003; 45: 173–180.

    Article  CAS  PubMed  Google Scholar 

  12. Taniyama Y et al. Local delivery of plasmid DNA into rat carotid artery using ultrasound. Circulation 2002; 105: 1233–1239.

    Article  CAS  PubMed  Google Scholar 

  13. Endoh M et al. Fetal gene transfer by intrauterine injection with microbubble-enhanced ultrasound. Mol Ther 2002; 5: 501–508.

    Article  CAS  PubMed  Google Scholar 

  14. Mychaskiw II G et al. Optison (FS069) disrupts the blood–brain barrier in rats. Anesth Analg 2000; 91: 798–803.

    Article  CAS  PubMed  Google Scholar 

  15. Hynynen K et al. The threshold for brain damage in rabbits induced by bursts of ultrasound in the presence of an ultrasound contrast agent (Optison). Ultrasound Med Biol 2003; 29: 473–481.

    Article  PubMed  Google Scholar 

  16. Cavaglia M et al. Regional variation in brain capillary density and vascular response to ischemia. Brain Res 2001; 910: 81–93.

    Article  CAS  PubMed  Google Scholar 

  17. Somia N, Verma IM . Gene therapy: trials and tribulations. Nat Rev Genet 2000; 1: 91–99.

    Article  CAS  PubMed  Google Scholar 

  18. Unger EC, Hersh E, Vannan M, McCreery T . Gene delivery using ultrasound contrast agents. Echocardiography 2001; 18: 355–361.

    Article  CAS  PubMed  Google Scholar 

  19. Taniyama Y et al. Development of safe and efficient novel nonviral gene transfer using ultrasound: enhancement of transfection efficiency of naked plasmid DNA in skeletal muscle. Gene Therapy 2002; 9: 372–380.

    Article  CAS  PubMed  Google Scholar 

  20. Tachibana K et al. Induction of cell-membrane porosity by ultrasound. Lancet 1999; 353: 1409.

    Article  CAS  PubMed  Google Scholar 

  21. Daffertshofer M, Fatar M . Therapeutic ultrasound in ischemic stroke treatment: experimental evidence. Eur J Ultrasound 2002; 16: 121–130.

    Article  PubMed  Google Scholar 

  22. Yukawa H et al. Adenoviral gene transfer of basic fibroblast growth factor promotes angiogenesis in rat brain. Gene Therapy 2000; 7: 942–949.

    Article  CAS  PubMed  Google Scholar 

  23. Shimamura M et al. HVJ-envelope vector for gene transfer into central nervous system. Biochem Biophys Res Commun 2003; 2: 464–471.

    Article  Google Scholar 

  24. Albrecht PJ et al. Ciliary neurotrophic factor activates spinal cord astrocytes, stimulating their production and release of fibroblast growth factor-2, to increase motor neuron survival. Exp Neurol 2002; 173: 46–62.

    Article  CAS  PubMed  Google Scholar 

  25. Gill SS et al. Direct brain infusion of glial cell line-derived neurotrophic factor in Parkinson disease. Nat Med 2003; 9: 589–595.

    Article  CAS  PubMed  Google Scholar 

  26. Kordower JH . In vivo gene delivery of glial cell line-derived neurotrophic factor for Parkinson's disease. Ann Neurol 2003; 53: S120–S132; discussion S132–S134.

    Article  CAS  PubMed  Google Scholar 

  27. Wang L et al. Delayed delivery of AAV-GDNF prevents nigral neurodegeneration and promotes functional recovery in a rat model of Parkinson's disease. Gene Therapy 2002; 9: 381–389.

    Article  CAS  PubMed  Google Scholar 

  28. Georgievska B et al. Neuroprotection in the rat Parkinson model by intrastriatal GDNF gene transfer using a lentiviral vector. Neuroreport 2002; 13: 75–82.

    Article  CAS  PubMed  Google Scholar 

  29. Behrens S et al. Transcranial ultrasound-improved thrombolysis: diagnostic versus therapeutic ultrasound. Ultrasound Med Biol 2001; 27: 1683–1689.

    Article  CAS  PubMed  Google Scholar 

  30. Nagai T et al. A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications. Nat Biotechnol 2002; 20: 87–90.

    Article  CAS  PubMed  Google Scholar 

  31. Tsujie M et al. Electroporation-mediated gene transfer that targets glomeruli. J Am Soc Nephrol 2001; 12: 949–954.

    CAS  PubMed  Google Scholar 

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Acknowledgements

This work was partially supported by a Grant-in-Aid from the Organization for Pharmaceutical Safety and Research, a Grant-in-Aid from The Ministry of Public Health and Welfare, a Grant-in-Aid from Japan Promotion of Science, and through Special Coordination Funds of the Ministry of Education, Culture, Sports, Science and Technology, the Japanese Government.

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Authors and Affiliations

  1. Division of Clinical Gene Therapy, Osaka University, Yamada-oka, Suita, Japan

    M Shimamura, N Sato, Y Taniyama & R Morishita

  2. Division of Gene Therapy Science, Osaka University, Yamada-oka, Suita, Japan

    M Shimamura, S Yamamoto, M Endoh & Y Kaneda

  3. Department of Geriatric Medicine, Graduate School of Medicine, Osaka University, Yamada-oka, Suita, Japan

    H Kurinami, M Aoki & T Ogihara

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  1. M Shimamura
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  2. N Sato
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Shimamura, M., Sato, N., Taniyama, Y. et al. Development of efficient plasmid DNA transfer into adult rat central nervous system using microbubble-enhanced ultrasound. Gene Ther 11, 1532–1539 (2004). https://doi.org/10.1038/sj.gt.3302323

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