Comparison of AAV Serotypes for Gene Delivery to Dopaminergic Neurons in the Substantia Nigra

Korecka, Joanna A.; Schouten, Marijn; Eggers, Ruben; Ulusoy, Ayşe; Bossers, Koen; Verhaagen, Joost

doi:10.5772/18939

Cited by 7 publications

(6 citation statements)

References 44 publications

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“…Efficient transduction of dopaminergic neurons in the SN with recombinant AAV serotypes 6, 8, and 9 has previously been reported (Klein et al 2006(Klein et al , 2008Dusonchet et al, 2009;Van der Perren et al, 2011). Comparison of AAV serotypes 5, 6, 7, and 8 for transduction of rat nigral dopaminergic neurons using synapsin promoter driven GFP reporter gene expression yielded no difference between these serotypes with regard to the percentage of GFP-expressing TH-positive neurons (Korecka et al, 2011). In a study by Klein et al (2008) investigating AAV transduction kinetics, the onset of transgene expression was found to be faster for AAV9 than for AAV8, as evidenced by the higher AAV9 transgene expression level, 1 and 2 weeks after viral injections to the SN.…”

Section: Discussionmentioning

confidence: 75%

Direct and Retrograde Transduction of Nigral Neurons with AAV6, 8, and 9 and Intraneuronal Persistence of Viral Particles

Löw

Aebischer²,

Schneider³

2013

Human Gene Therapy

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Recombinant adeno-associated viral (AAV) vectors of serotypes 6, 8, and 9 were characterized as tools for gene delivery to dopaminergic neurons in the substantia nigra for future gene therapeutic applications in Parkinson's disease. While vectors of all three serotypes transduced nigral dopaminergic neurons with equal efficiency when directly injected to the substantia nigra, AAV6 was clearly superior to AAV8 and AAV9 for retrograde transduction of nigral neurons after striatal delivery. For sequential transduction of nigral dopaminergic neurons, the combination of AAV9 with AAV6 proved to be more powerful than AAV8 with AAV6 or repeated AAV6 administration. Surprisingly, single-stranded viral genomes persisted in nigral dopaminergic neurons within cell bodies and axon terminals in the striatum, and intact assembled AAV capsid was enriched in nuclei of nigral neurons, 4 weeks after virus injections to the substantia nigra. 6-Hydroxydopamine (6-OHDA)-induced degeneration of dopaminergic neurons in the substantia nigra reduced the number of viral genomes in the striatum, in line with viral genome persistence in axon terminals. However, 6-OHDA-induced axonal degeneration did not induce any transsynaptic spread of AAV infection in the striatum. Therefore, the potential presence of viral particles in axons may not represent an important safety issue for AAV gene therapy applications in neurodegenerative diseases.

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Section: Discussionmentioning

confidence: 75%

Direct and Retrograde Transduction of Nigral Neurons with AAV6, 8, and 9 and Intraneuronal Persistence of Viral Particles

Löw

Aebischer²,

Schneider³

2013

Human Gene Therapy

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“…AAV-mPGK-eGFP was made by amplifying the eGFP sequence of AAV-hCMV-eGFP by using the primers 5′ GGAATTC ATGGTGAGCAAGGGCGAG 3′ and 5′ AGCGCTTTA CTTGTACAGCTCGTCCATG 3′ and was then cloned into an AAV-mPGK plasmid, which was a gift from Patrick Aebischer (Addgene plasmid # 24593), via HindIII-HF (NEB, R3104) and EcoRI-HF (NEB, R3101) digestion. AAV-hSYN-eGFP was made by removing the WPRE from plasmid pTRUF20B-SEW, a gift from Deniz Kirik (Lund University) described in [25], by HincII (NEB, R0103) digestion.…”

Section: Aav Vector Plasmidsmentioning

confidence: 99%

Optimization of adeno-associated viral vector-mediated transduction of the corticospinal tract: comparison of four promoters

et al. 2020

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Adeno-associated viral vectors are widely used as vehicles for gene transfer to the nervous system. The promoter and viral vector serotype are two key factors that determine the expression dynamics of the transgene. A previous comparative study has demonstrated that AAV1 displays efficient transduction of layer V corticospinal neurons, but the optimal promoter for transgene expression in corticospinal neurons has not been determined yet. In this paper, we report a side-by-side comparison between four commonly used promoters: the short CMV early enhancer/chicken β actin (sCAG), human cytomegalovirus (hCMV), mouse phosphoglycerate kinase (mPGK) and human synapsin (hSYN) promoter. Reporter constructs with each of these promoters were packaged in AAV1, and were injected in the sensorimotor cortex of rats and mice in order to transduce the corticospinal tract. Transgene expression levels and the cellular transduction profile were examined after 6 weeks. The AAV1 vectors harbouring the hCMV and sCAG promoters resulted in transgene expression in neurons, astrocytes and oligodendrocytes. The mPGK and hSYN promoters directed the strongest transgene expression. The mPGK promoter did drive expression in cortical neurons and oligodendrocytes, while transduction with AAV harbouring the hSYN promoter resulted in neuron-specific expression, including perineuronal net expressing interneurons and layer V corticospinal neurons. This promoter comparison study contributes to improve transgene delivery into the brain and spinal cord. The optimized transduction of the corticospinal tract will be beneficial for spinal cord injury research.

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“…The animals were placed in a stereotaxic frame (Kopf), where a 10 μl Hamilton syringe and 31 gauge needles were used as a delivery system. All injections were made into the substantia nigra pars compacta using the following anteroposterior (AP), mediolateral (ML), and dorsoventral (DV) coordinates as previously verified [23,54]: AP − 5.2 mm, ML − 2.0 mm, DV − 7.2 mm. The injection titers of the viruses were 2 × 10 12 vgc/mL for aSYN, and 8.7 × 10 11 vgc/mL for HEX A, B, and EV, and 3 μl were injected per site.…”

Section: Stereotactic Delivery Of Aav6 To the Rat Substantia Nigra Pamentioning

confidence: 99%

Upregulating β-hexosaminidase activity in rodents prevents α-synuclein lipid associations and protects dopaminergic neurons from α-synuclein-mediated neurotoxicity

Brekk

Korecka

Crapart

et al. 2020

acta neuropathol commun

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Sandhoff disease (SD) is a lysosomal storage disease, caused by loss of β-hexosaminidase (HEX) activity resulting in the accumulation of ganglioside GM2. There are shared features between SD and Parkinson's disease (PD). α-synuclein (aSYN) inclusions, the diagnostic hallmark sign of PD, are frequently found in the brain in SD patients and HEX knockout mice, and HEX activity is reduced in the substantia nigra in PD. In this study, we biochemically demonstrate that HEX deficiency in mice causes formation of high-molecular weight (HMW) aSYN and ubiquitin in the brain. As expected from HEX enzymatic function requirements, overexpression in vivo of HEXA and B combined, but not either of the subunits expressed alone, increased HEX activity as evidenced by histochemical assays. Biochemically, such HEX gene expression resulted in increased conversion of GM2 to its breakdown product GM3. In a neurodegenerative model of overexpression of aSYN in rats, increasing HEX activity by AAV6 gene transfer in the substantia nigra reduced aSYN embedding in lipid compartments and rescued dopaminergic neurons from degeneration. Overall, these data are consistent with a paradigm shift where lipid abnormalities are central to or preceding protein changes typically associated with PD.

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Comparison of AAV Serotypes for Gene Delivery to Dopaminergic Neurons in the Substantia Nigra

Cited by 7 publications

References 44 publications

Direct and Retrograde Transduction of Nigral Neurons with AAV6, 8, and 9 and Intraneuronal Persistence of Viral Particles

Direct and Retrograde Transduction of Nigral Neurons with AAV6, 8, and 9 and Intraneuronal Persistence of Viral Particles

Optimization of adeno-associated viral vector-mediated transduction of the corticospinal tract: comparison of four promoters

Upregulating β-hexosaminidase activity in rodents prevents α-synuclein lipid associations and protects dopaminergic neurons from α-synuclein-mediated neurotoxicity

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