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Liposome-mediated transfer of the bcl-2 gene results in neuroprotection after in vivo transient focal cerebral ischemia in an animal model

Gene Therapy volume 9, pages 415–419 (2002)Cite this article

Abstract

Acute cerebral ischemia causes hypoxic neuronal cell death by necrosis and apoptosis. Expression of anti-apoptotic transgenes in ischemic brain may provide a useful therapeutic strategy for alleviation of postischemic damage. The present study investigates liposome-mediated transfer of the human bcl-2 protein in a rat model of focal transient ischemia due to middle cerebral artery (MCA) occlusion. Two different types of plasmid vectors were used for bcl-2 expression: one driven by the constitutive cytomegalovirus promoter (pCMV) and another based on the hypoxia-inducible human vascular endothelial growth factor promoter (pHRE). Cationic liposome/plasmid DNA complexes (lipoplexes) were injected directly into the cerebrospinal fluid (CSF) of rats immediately after MCA occlusion. The brains of treated and control animals were analyzed 48 h later. Infarct volumes and numbers of apoptotic cells were quantified. Occlusion of the MCA resulted in ipsilateral cerebral infarcts in all study animals. Transfer of the bcl-2 gene resulted in high level widespread protein expression in the case of the pCMV-bcl2 plasmid, while animals treated with the pHRE-bcl2 vector showed lower expression levels of bcl2 which were in addition limited to the ischemic area. Treatment with pCMV-bcl2, but not with pHRE-bcl2, was able to significantly reduce the infarct volume, which was 109 ± 8 mm3 for pCMV-bcl2, 152 ± 29 mm3 for pHRE-bcl2, and 155 ± 18 mm3 for control animals. Animals transfected with either vector showed a significant reduction in numbers of apoptotic cells in the infarct and penumbra area compared with controls. There were no short-term neurological side-effects of the CSF injection of lipoplexes or of bcl-2 expression. In conclusion, the hypoxia-inducible bcl-2 expression mediated by intrathecal lipoplexes may represent a novel, biologically safe and lesion-selective therapeutic approach for neuroprotection after acute cerebral ischemia.

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Acknowledgements

This study was supported by a grant from the E-Kröner-Fresenius Foundation (NGR), and in parts by grants Ra 705/4-1 from the German Research Council (DFG) and 2794A/0087H and 3067A/0029G from the Federal State of Saxony-Anhalt, and a grant from the Bartling Foundation Hannover (all to NGR).

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Authors and Affiliations

  1. Department of Neurosurgery, Molecular Neuro-oncology Laboratory, Martin-Luther-University, Halle/S, Germany

    Y-J Cao

  2. Department of Radiation Oncology, Mayer Cancer Biology Research Laboratory, Stanford University School of Medicine, Stanford, CA, USA

    T Shibata

  3. Department of Experimental Oncology, Research Laboratories of Schering AG, Berlin, Germany

    Y-J Cao

  4. Department of Neurological Science, University of Liverpool, Liverpool, UK

    NG Rainov

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  1. Y-J Cao
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  2. T Shibata
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Cao, YJ., Shibata, T. & Rainov, N. Liposome-mediated transfer of the bcl-2 gene results in neuroprotection after in vivo transient focal cerebral ischemia in an animal model. Gene Ther 9, 415–419 (2002). https://doi.org/10.1038/sj.gt.3301676

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