Optimizing AAV2/6 microglial targeting identified enhanced efficiency in the photoreceptor degenerative environment
| Autor: | Raquel Casado-Polanco, Gloria Colombo, Gabriele M. Wögenstein, Margaret E. Maes, Sandra Siegert |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Cell type
retina Transgene viruses in vivo transduction Outer plexiform layer microglia degeneration adeno-associated virus (AAV) QH426-470 03 medical and health sciences Transduction (genetics) 0302 clinical medicine medicine Genetics Molecular Biology 030304 developmental biology 0303 health sciences Retina Microglia QH573-671 Chemistry Point mutation Cell biology medicine.anatomical_structure Capsid Molecular Medicine Original Article Cytology 030217 neurology & neurosurgery |
| Zdroj: | Molecular Therapy. Methods & Clinical Development Molecular Therapy: Methods & Clinical Development, Vol 23, Iss, Pp 210-224 (2021) Molecular Therapy-Methods & Clinical Development |
| ISSN: | 2329-0501 |
| Popis: | Adeno-associated viruses (AAVs) are widely used to deliver genetic material in vivo to distinct cell types such as neurons or glial cells, allowing for targeted manipulation. Transduction of microglia is mostly excluded from this strategy, likely due to the cells’ heterogeneous state upon environmental changes, which makes AAV design challenging. Here, we established the retina as a model system for microglial AAV validation and optimization. First, we show that AAV2/6 transduced microglia in both synaptic layers, where layer preference corresponds to the intravitreal or subretinal delivery method. Surprisingly, we observed significantly enhanced microglial transduction during photoreceptor degeneration. Thus, we modified the AAV6 capsid to reduce heparin binding by introducing four point mutations (K531E, R576Q, K493S, and K459S), resulting in increased microglial transduction in the outer plexiform layer. Finally, to improve microglial-specific transduction, we validated a Cre-dependent transgene delivery cassette for use in combination with the Cx3cr1CreERT2 mouse line. Together, our results provide a foundation for future studies optimizing AAV-mediated microglia transduction and highlight that environmental conditions influence microglial transduction efficiency. Graphical abstract Targeting microglia in vivo using AAVs is neither robust nor reliable. Here, Maes et al. show that transduction of retinal microglia niches depends on the injection method and the degeneration environment. This informed development of a capsid-modified AAV2/6 that in combination with the Cre-loxP system improved microglia specificity. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|