Tackling obstacles for gene therapy targeting neurons: disrupting perineural nets with hyaluronidase improves transduction
| Autor: | Stjepana Kovac, Klaus Wanisch, Stephanie Schorge |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Nervous system
Male Pathology lcsh:Medicine Biochemistry Extracellular matrix Rats Sprague-Dawley Transduction (genetics) chemistry.chemical_compound 0302 clinical medicine Transduction Genetic Molecular Cell Biology Image Processing Computer-Assisted lcsh:Science Neurons 0303 health sciences Multidisciplinary Neurogenesis Brain Transfection Genomics Gene Therapy Cell biology Extracellular Matrix medicine.anatomical_structure Cytochemistry Medicine Viral Vectors Cellular Types Research Article medicine.medical_specialty Genetic Vectors Primary Cell Culture Hyaluronoglucosaminidase Biology Microbiology Viral vector 03 medical and health sciences Genomic Medicine In vivo Virology medicine Animals Humans Propidium iodide 030304 developmental biology Clinical Genetics Lentivirus lcsh:R Genetic Therapy Rats HEK293 Cells chemistry Cellular Neuroscience lcsh:Q Molecular Neuroscience Nerve Net 030217 neurology & neurosurgery Viral Transmission and Infection Neuroscience |
| Zdroj: | PLoS ONE, Vol 8, Iss 1, p e53269 (2013) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Gene therapy has been proposed for many diseases in the nervous system. In most cases for successful treatment, therapeutic vectors must be able to transduce mature neurons. However, both in vivo, and in vitro, where preliminary characterisation of viral particles takes place, transduction of neurons is typically inefficient. One possible explanation is that the extracellular matrix (ECM), forming dense perineural nets (PNNs) around neurons, physically blocks access to the cell surface. We asked whether co-administration of lentiviral vectors with an enzyme that disrupts the ECM could improve transduction efficiency. Using hyaluronidase, an enzyme which degrades hyaluronic acid, a high molecular weight molecule of the ECM with mainly a scaffolding function, we show that in vitro in mixed primary cortical cultures, and also in vivo in rat cortex, hyaluronidase co-administration increased the percentage of transduced mature, NeuN-positive neurons. Moreover, hyaluronidase was effective at doses that showed no toxicity in vitro based on propidium iodide staining of treated cultures. Our data suggest that limited efficacy of neuronal transduction is partly due to PNNs surrounding neurons, and further that co-applying hyaluronidase may benefit applications where efficient transduction of neurons in vitro or in vivo is required. |
| Databáze: | OpenAIRE |
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