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Neuraminidase-mediated desialylation augments AAV9-mediated gene expression in skeletal muscle.
J Gene Med 2018; 20(9):e3049JG

Abstract

BACKGROUND

Following systemic delivery, AAV9-mediated gene expression is significantly increased in ischemic versus non-ischemic muscle, suggesting that AAV9 is an attractive vector for treating peripheral arterial disease. Potential mechanisms underlying ischemia-augmented expression include: (i) increased vascular permeability and (ii) "unmasking" of endogenous AAV9 receptors. In the present study, we aimed to reconstitute the ischemic induction of AAV9 in vivo, using local injection of histamine (to increase vascular permeability) and neuraminidase (to desialylate cell surface glycans).

METHODS

Bioassays were performed to optimize the effects of histamine and neuraminidase after intramuscular injection. Histamine and/or neuraminidase were then injected intramuscularly shortly before intravenous injection of an AAV9 vector expressing luciferase. Luciferase expression was serially assessed with bioluminescence imaging. At the end of the study, tissues were harvested for assays of luciferase activity and AAV9 genome copy number aiming to assess AAV-mediated gene expression and transduction, respectively.

RESULTS

Intramuscular injection of either neuraminidase or neuraminidase plus histamine significantly increased both transduction and gene expression, whereas histamine alone had little effect. Pre-injection with neuraminidase increased AAV9-mediated gene delivery by four- to nine-fold and luciferase activity by 60-100-fold. Luciferase activity in neuraminidase-injected muscle was > 100-fold higher than in any off-target tissue (including heart, liver and brain).

CONCLUSIONS

The ischemic induction of AAV9-mediated gene expression in muscle can largely be reconstituted by pre-injecting neuraminidase intranmuscularly. This strategy may prove useful in future human gene therapy protocols as a quick and efficient means to selectively target systemically injected AAV9 to localized regions of muscle, thus decreasing the potential for adverse effects in off-target tissues.

Authors+Show Affiliations

Department of Medicine, Division of Cardiovascular Medicine, University of Virginia, Charlottesville, VA, USA.Department of Medicine, Division of Cardiovascular Medicine, University of Virginia, Charlottesville, VA, USA.Department of Medicine, Division of Cardiovascular Medicine, University of Virginia, Charlottesville, VA, USA.Department of Medicine, Division of Cardiovascular Medicine, University of Virginia, Charlottesville, VA, USA. Department of Biomedical Engineering, University of Virginia, Charlottesville, VA, USA. Robert M. Berne Cardiovascular Research Center, University of Virginia, Charlottesville, VA, USA.Department of Medicine, Division of Cardiovascular Medicine, University of Virginia, Charlottesville, VA, USA. Department of Biomedical Engineering, University of Virginia, Charlottesville, VA, USA. Robert M. Berne Cardiovascular Research Center, University of Virginia, Charlottesville, VA, USA.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

30101537

Citation

Zhu, Hongling, et al. "Neuraminidase-mediated Desialylation Augments AAV9-mediated Gene Expression in Skeletal Muscle." The Journal of Gene Medicine, vol. 20, no. 9, 2018, pp. e3049.
Zhu H, Wang T, John Lye R, et al. Neuraminidase-mediated desialylation augments AAV9-mediated gene expression in skeletal muscle. J Gene Med. 2018;20(9):e3049.
Zhu, H., Wang, T., John Lye, R., French, B. A., & Annex, B. H. (2018). Neuraminidase-mediated desialylation augments AAV9-mediated gene expression in skeletal muscle. The Journal of Gene Medicine, 20(9), pp. e3049. doi:10.1002/jgm.3049.
Zhu H, et al. Neuraminidase-mediated Desialylation Augments AAV9-mediated Gene Expression in Skeletal Muscle. J Gene Med. 2018;20(9):e3049. PubMed PMID: 30101537.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Neuraminidase-mediated desialylation augments AAV9-mediated gene expression in skeletal muscle. AU - Zhu,Hongling, AU - Wang,Tao, AU - John Lye,Robert, AU - French,Brent A, AU - Annex,Brian H, Y1 - 2018/09/04/ PY - 2018/01/23/received PY - 2018/08/01/revised PY - 2018/08/01/accepted PY - 2018/8/14/pubmed PY - 2018/8/14/medline PY - 2018/8/14/entrez KW - Adeno-associated virus KW - gene therapy KW - muscle-specific promoter KW - neuraminidase KW - receptor SP - e3049 EP - e3049 JF - The journal of gene medicine JO - J Gene Med VL - 20 IS - 9 N2 - BACKGROUND: Following systemic delivery, AAV9-mediated gene expression is significantly increased in ischemic versus non-ischemic muscle, suggesting that AAV9 is an attractive vector for treating peripheral arterial disease. Potential mechanisms underlying ischemia-augmented expression include: (i) increased vascular permeability and (ii) "unmasking" of endogenous AAV9 receptors. In the present study, we aimed to reconstitute the ischemic induction of AAV9 in vivo, using local injection of histamine (to increase vascular permeability) and neuraminidase (to desialylate cell surface glycans). METHODS: Bioassays were performed to optimize the effects of histamine and neuraminidase after intramuscular injection. Histamine and/or neuraminidase were then injected intramuscularly shortly before intravenous injection of an AAV9 vector expressing luciferase. Luciferase expression was serially assessed with bioluminescence imaging. At the end of the study, tissues were harvested for assays of luciferase activity and AAV9 genome copy number aiming to assess AAV-mediated gene expression and transduction, respectively. RESULTS: Intramuscular injection of either neuraminidase or neuraminidase plus histamine significantly increased both transduction and gene expression, whereas histamine alone had little effect. Pre-injection with neuraminidase increased AAV9-mediated gene delivery by four- to nine-fold and luciferase activity by 60-100-fold. Luciferase activity in neuraminidase-injected muscle was > 100-fold higher than in any off-target tissue (including heart, liver and brain). CONCLUSIONS: The ischemic induction of AAV9-mediated gene expression in muscle can largely be reconstituted by pre-injecting neuraminidase intranmuscularly. This strategy may prove useful in future human gene therapy protocols as a quick and efficient means to selectively target systemically injected AAV9 to localized regions of muscle, thus decreasing the potential for adverse effects in off-target tissues. SN - 1521-2254 UR - https://www.unboundmedicine.com/medline/citation/30101537/Neuraminidase_mediated_desialylation_augments_AAV9_mediated_gene_expression_in_skeletal_muscle_ L2 - https://doi.org/10.1002/jgm.3049 DB - PRIME DP - Unbound Medicine ER -