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Decline in Exogenous Gene Expression in Primate Brain Following Intravenous Administration Is Due to Plasmid Degradation

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Purpose

Nonviral gene transfer to the brain of adult Rhesus monkeys is possible with a single intravenous administration of plasmid DNA that is encapsulated in the interior of pegylated immunoliposomes, which are targeted across membrane barriers in vivo with a monoclonal antibody to the human insulin receptor.

Methods

The present studies measure the rate of decay of luciferase gene expression in the Rhesus monkey with luciferase enzyme assays, Southern blotting, and real-time polymerase chain reaction.

Results

Luciferase enzyme activity in frontal cortex, cerebellum, and liver decays with a t1/2 of 2.1 ± 0.1, 2.6 ± 0.2, and 1.7 ± 0.01 days, respectively. Luciferase plasmid in brain and liver was detectable by Southern blotting at 2 days, but not at 7 or 14 days. The concentration of luciferase plasmid DNA in brain and liver was measured by real-time polymerase chain reaction, and decayed with t1/2 of 1.3 ± 0.3 and 2.7 ± 0.5 days, respectively.

Conclusions

The maximal concentration of luciferase plasmid DNA in Rhesus monkey brain was 3–4 molecules/cell following an i.v. administration of 12 μg/kg pegylated immunoliposome encapsulated plasmid DNA. These results demonstrate that the rate of loss of exogenous gene expression in the primate in vivo correlates with the rate of DNA degradation of the exogenous plasmid DNA.

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Acknowledgments

This work was supported by a grant from the Neurotoxin Exposure Treatment Research Program of the U.S. Department of Defense, and by NIH grant NS-53540.

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Authors and Affiliations

  1. Department of Medicine, UCLA, UCLA Warren Hall 13-164, 900 Veteran Avenue, Los Angeles, CA, 90024, USA

    Chun Chu, Yun Zhang, Ruben J. Boado & William M. Pardridge

Authors
  1. Chun Chu
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  2. Yun Zhang
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  3. Ruben J. Boado
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  4. William M. Pardridge
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Corresponding author

Correspondence to William M. Pardridge.

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Chu, C., Zhang, Y., Boado, R.J. et al. Decline in Exogenous Gene Expression in Primate Brain Following Intravenous Administration Is Due to Plasmid Degradation. Pharm Res 23, 1586–1590 (2006). https://doi.org/10.1007/s11095-006-0274-x

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  • DOI: https://doi.org/10.1007/s11095-006-0274-x

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