Intrathecal Injection of Naked Plasmid DNA Provides Long-term Expression of Secreted Proteins
| Autor: | Kirk W. Johnson, Erin D. Milligan, Stephen J. Langer, Leslie A. Leinwand, Linda R. Watkins, Travis S. Hughes, Raymond A. Chavez |
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| Rok vydání: | 2009 |
| Předmět: |
Male
Transgene Biology Rats Sprague-Dawley 03 medical and health sciences 0302 clinical medicine Plasmid Gene expression Drug Discovery Genetics Animals Humans Scaffold/matrix attachment region Molecular Biology Injections Spinal 030304 developmental biology Pharmacology 0303 health sciences Original Articles Alkaline Phosphatase Molecular biology Interleukin-10 Rats Secretory protein CpG site Naked DNA Alkaline phosphatase Molecular Medicine 030217 neurology & neurosurgery Plasmids |
| Zdroj: | Molecular Therapy. 17(1):88-94 |
| ISSN: | 1525-0016 |
| DOI: | 10.1038/mt.2008.230 |
| Popis: | Therapeutic benefit has been reported to result from intrathecal (i.t.) injection of transgene vectors, including naked DNA. However, most studies using naked DNA have measured only the transgene expression of intracellular proteins. Here we demonstrate that i.t. injection of naked DNA can result in long-term expression of secreted proteins. Plasmids expressing either secreted alkaline phosphatase (SEAP) or human interleukin-10 (hIL-10) were injected into the i.t. space in rats, and transgene products were repeatedly measured in the cerebrospinal fluid (CSF). Both SEAP and hIL-10 were maximal at 1 and 2 days after the injection and still detectable at 4 months. The utilization of a plasmid having two features that are hypothesized to increase gene expression (matrix attachment regions (MARs) and lack of CpG dinucleotides) resulted in a significant increase in gene expression. Reinjection of SEAP or hIL-10 plasmids after 4 months significantly increased protein levels at 1 and 14 days after the reinjection. SEAP was uniformly distributed between the DNA delivery site (approximately vertebral level T13) and the lumbar puncture site (L5/L6 inter-vertebral space), was reduced at the cisterna magna, and was detectable, though at much lower levels, in serum. These data suggest that naked DNA has the potential to be used as a therapeutic tool for applications that require long-term release of transgenes into the CSF. |
| Databáze: | OpenAIRE |
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