Immunogenicity of intrathecal plasmid gene delivery: cytokine release and effects on transgene expression
Open accessAbstract
BACKGROUND: One method for the delivery of therapeutic proteins to the spinal cord is to inject nonviral gene vectors including plasmid DNA into the cerebrospinal fluid (CSF) that surrounds the spinal cord (intrathecal space). This approach has produced therapeutic benefits in animal models of disease and several months of protein expression; however, there is little information available on the immune response to these treatments in the intrathecal space, the relevance of plasmid CpG sequences to any plasmid-induced immune response, or the effect of this immune response on transgene expression. METHODS: In the present study, coding or noncoding plasmids were delivered to the intrathecal space of the lumbar spinal region in rats. Lumbosacral CSF was then collected at various time points afterwards for monitoring of cytokines and transgene expression. RESULTS: This work demonstrates, for the first time, increased tumor necrosis factor-alpha and interleukin-1 in response to intrathecal plasmid vector injection and provides evidence indicating that this response is largely absent in a CpG-depleted vector. Transgene expression in the CSF is not significantly affected by this immune response. Expression after intrathecal plasmid injection is variable across rats but correlates with the amount of tissue associated plasmid and is increased by disrupting normal CSF flow. CONCLUSIONS: The data obtained in the present study indicate that plasmid immunogenicity may affect intrathecal plasmid gene therapy safety but not transgene expression in the CSF. Furthermore, the development of methods to prevent loss of plasmid via CSF flow out of the central nervous system through the injection hole and/or natural outflow routes may increase intrathecal plasmid gene delivery efficacy.
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Plasmid-based expression often leads to a longer duration of Cas9 protein, which may result in higher off-target effects and strong immune responses.
“In addition, the plasmid-based expression often leads to a longer duration of Cas9 protein, which may result in higher off-target effects and strong immune responses [95].”
From Non-viral delivery systems for CRISPR/Cas9-based genome editing: challenges and opportunities (2018) · cites this paper as [95]
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- Authors
- Hughes, T. S., Langer, S. J., Virtanen, S. I., Chavez, R. A., Watkins, L. R., Milligan, E. D., & Leinwand, L. A.
- Affiliations
- University of Colorado Boulder (US)
- University of New Mexico (US)
- Corresponding author
- Leinwand, Leslie A.
- ORCID
- Venue
- The Journal of Gene Medicine
- Publisher
- Wiley
- Published
- 2009-06-16
- Volume
- 11
- Issue
- 9
- Pages
- 782-790
- ISSN
- 1099-498X (ISSN-L)
- 1521-2254
- PMC
- PMC3649870
- OpenAlex
- W1990187837
- Cited by
- 28
- References
- 38
- Citations by year
- 2026: 2 · 2025: 4 · 2024: 3 · 2023: 3 · 2022: 2 · 2021: 2 · 2020: 2 · 2019: 2 · 2018: 3 · 2017: 1 · 2016: 1 · 2014: 1 · 2012: 1
- FWCI
- 0.27
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- The Journal of Gene Medicine (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health
- National Institute on Drug Abuse (K02DA015642, DA024044, DA 015642, R01DA018156, K05DA024044, DA 018156)
- National Institute of General Medical Sciences (GM 29090, R01GM029090)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
- Keywords
- intrathecal gene delivery, plasmid DNA, immunogenicity, cytokine production, TNF-α, interleukin-1, CpG sequences, transgene expression, cerebrospinal fluid, lumbar spinal cord, cerebrospinal fluid flow, non-viral vectors, gene therapy safety
- MeSH terms
- Animals, Cell Line, Humans, Injections, Spinal, Male, Plasmids, Transfection, Genetic Therapy, Gene Expression, Cytokines, Rats, Sprague-Dawley, CpG Islands, Transgenes, Toll-Like Receptor 9, Rats
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