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AAV9 supports wide-scale transduction of the CNS and TDP-43 disease modeling in adult rats.

Jackson KL, Dayton RD, Klein RL - Mol Ther Methods Clin Dev (2015)

Bottom Line: AAV9 has emerged as an efficient adeno-associated virus (AAV) serotype for gene transfer to the central nervous system.In order to optimize the gene transfer, we made comparisons of efficiency by age, gender, and across several AAV serotypes (AAV1, AAV8, AAV9, and AAV10).The data indicate more efficient neuronal transduction in neonates, with little evidence of glial transduction at either age, no gender-related differences in transduction, and that AAV9 was efficient in adults relative to the other serotypes tested.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Toxicology, and Neuroscience, Louisiana State University Health Sciences Center , Shreveport, Louisiana, USA.

ABSTRACT
AAV9 has emerged as an efficient adeno-associated virus (AAV) serotype for gene transfer to the central nervous system. We have used this technique to study aspects of amyotrophic lateral sclerosis (ALS) by administering AAV encoding the ALS-related gene transactive response DNA binding protein of 43 kDa (TDP-43) to neonatal rats. However, inducing the expression in adult subjects would be preferable to mimic the adult onset of symptoms in ALS. We expressed either green fluorescent protein (GFP) or TDP-43 in adult rats after an intravenous (i.v.) route of administration to attempt wide-scale transduction of the spinal cord for disease modeling. In order to optimize the gene transfer, we made comparisons of efficiency by age, gender, and across several AAV serotypes (AAV1, AAV8, AAV9, and AAV10). The data indicate more efficient neuronal transduction in neonates, with little evidence of glial transduction at either age, no gender-related differences in transduction, and that AAV9 was efficient in adults relative to the other serotypes tested. Based on these data, AAV9 TDP-43 was expressed at three vector doses in adult female rats yielding highly consistent, dose-dependent motor deficits. AAV9 can be delivered i.v. to adult rats to achieve consistent pathophysiological changes and a relevant adult-onset system for disease modeling.

No MeSH data available.


Related in: MedlinePlus

AAV9 GFP gene transfer in females and males after i.v. vector administration to either (a, b) neonates or (c, d) adults. In the spinal cord, the percentage of motor neurons expressing GFP was estimated. In the cerebellum, an imaging program was used to estimate the intensity of GFP immunoreactivity (Supplementary Figure S1). There was no significant effect of gender on transduction efficiency using AAV9 at either age (N = 4–9/group). GFP, green fluorescent protein; i.v., intravenous.
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fig4: AAV9 GFP gene transfer in females and males after i.v. vector administration to either (a, b) neonates or (c, d) adults. In the spinal cord, the percentage of motor neurons expressing GFP was estimated. In the cerebellum, an imaging program was used to estimate the intensity of GFP immunoreactivity (Supplementary Figure S1). There was no significant effect of gender on transduction efficiency using AAV9 at either age (N = 4–9/group). GFP, green fluorescent protein; i.v., intravenous.

Mentions: Potential effects of gender on AAV transduction efficiency were evaluated in both neonatal and adult rats using AAV9. There have been reports of sex hormone involvement in AAV transduction efficiency.20–22 However, in this study, there was no significant difference in transduction efficiency between genders at either the neonatal (Figure 4a,b) or adult age (Figure 4c,d). However, it is worth mentioning a trend of increased expression in adult female rats relative to adult male rats (P = 0.09; t-test), which would be consistent with the increased expression seen in adult female mice by Maguire et al.20 Each data point is shown to depict the variability of the approach, which was considerable in the neonatal males and the adult females as shown in Figure 4.


AAV9 supports wide-scale transduction of the CNS and TDP-43 disease modeling in adult rats.

Jackson KL, Dayton RD, Klein RL - Mol Ther Methods Clin Dev (2015)

AAV9 GFP gene transfer in females and males after i.v. vector administration to either (a, b) neonates or (c, d) adults. In the spinal cord, the percentage of motor neurons expressing GFP was estimated. In the cerebellum, an imaging program was used to estimate the intensity of GFP immunoreactivity (Supplementary Figure S1). There was no significant effect of gender on transduction efficiency using AAV9 at either age (N = 4–9/group). GFP, green fluorescent protein; i.v., intravenous.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4588447&req=5

fig4: AAV9 GFP gene transfer in females and males after i.v. vector administration to either (a, b) neonates or (c, d) adults. In the spinal cord, the percentage of motor neurons expressing GFP was estimated. In the cerebellum, an imaging program was used to estimate the intensity of GFP immunoreactivity (Supplementary Figure S1). There was no significant effect of gender on transduction efficiency using AAV9 at either age (N = 4–9/group). GFP, green fluorescent protein; i.v., intravenous.
Mentions: Potential effects of gender on AAV transduction efficiency were evaluated in both neonatal and adult rats using AAV9. There have been reports of sex hormone involvement in AAV transduction efficiency.20–22 However, in this study, there was no significant difference in transduction efficiency between genders at either the neonatal (Figure 4a,b) or adult age (Figure 4c,d). However, it is worth mentioning a trend of increased expression in adult female rats relative to adult male rats (P = 0.09; t-test), which would be consistent with the increased expression seen in adult female mice by Maguire et al.20 Each data point is shown to depict the variability of the approach, which was considerable in the neonatal males and the adult females as shown in Figure 4.

Bottom Line: AAV9 has emerged as an efficient adeno-associated virus (AAV) serotype for gene transfer to the central nervous system.In order to optimize the gene transfer, we made comparisons of efficiency by age, gender, and across several AAV serotypes (AAV1, AAV8, AAV9, and AAV10).The data indicate more efficient neuronal transduction in neonates, with little evidence of glial transduction at either age, no gender-related differences in transduction, and that AAV9 was efficient in adults relative to the other serotypes tested.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Toxicology, and Neuroscience, Louisiana State University Health Sciences Center , Shreveport, Louisiana, USA.

ABSTRACT
AAV9 has emerged as an efficient adeno-associated virus (AAV) serotype for gene transfer to the central nervous system. We have used this technique to study aspects of amyotrophic lateral sclerosis (ALS) by administering AAV encoding the ALS-related gene transactive response DNA binding protein of 43 kDa (TDP-43) to neonatal rats. However, inducing the expression in adult subjects would be preferable to mimic the adult onset of symptoms in ALS. We expressed either green fluorescent protein (GFP) or TDP-43 in adult rats after an intravenous (i.v.) route of administration to attempt wide-scale transduction of the spinal cord for disease modeling. In order to optimize the gene transfer, we made comparisons of efficiency by age, gender, and across several AAV serotypes (AAV1, AAV8, AAV9, and AAV10). The data indicate more efficient neuronal transduction in neonates, with little evidence of glial transduction at either age, no gender-related differences in transduction, and that AAV9 was efficient in adults relative to the other serotypes tested. Based on these data, AAV9 TDP-43 was expressed at three vector doses in adult female rats yielding highly consistent, dose-dependent motor deficits. AAV9 can be delivered i.v. to adult rats to achieve consistent pathophysiological changes and a relevant adult-onset system for disease modeling.

No MeSH data available.


Related in: MedlinePlus