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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

GFP expressed by AAV1, AAV8, AAV9, or AAV10 after i.v. vector administration to either neonates or adults. The GFP expression in the CNS appeared to be neuronal with any of the serotypes tested at either age. The different serotypes are shown in the different columns, and the spinal cord and cerebellum are shown at each age. The serotypes performed similarly for GFP expression in the neonates, but in adults, AAV9 appeared to result in the strongest GFP expression. Quantifications of the GFP expression are in Figure 6. Bar in a = 134 μm. Same magnification in b–h. Bar in i = 268 μm. Same magnification in j–l. Bar in m = 536 μm. Same magnification in n–p. CNS, central nervous system; GFP, green fluorescent protein; i.v., intravenous.
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fig5: GFP expressed by AAV1, AAV8, AAV9, or AAV10 after i.v. vector administration to either neonates or adults. The GFP expression in the CNS appeared to be neuronal with any of the serotypes tested at either age. The different serotypes are shown in the different columns, and the spinal cord and cerebellum are shown at each age. The serotypes performed similarly for GFP expression in the neonates, but in adults, AAV9 appeared to result in the strongest GFP expression. Quantifications of the GFP expression are in Figure 6. Bar in a = 134 μm. Same magnification in b–h. Bar in i = 268 μm. Same magnification in j–l. Bar in m = 536 μm. Same magnification in n–p. CNS, central nervous system; GFP, green fluorescent protein; i.v., intravenous.

Mentions: Next, we compared several AAV serotypes that have been used extensively in animals before, such as AAV1, AAV8, AAV9, and AAV10. The same GFP expression plasmid DNA was packaged into each type, and the same dose of viral particles were administered. GFP fluorescence was evaluated in the cerebellum and spinal cord 4 weeks post-administration. With neonatal administrations, we found no significant differences in transduction efficiency in the cerebellum or spinal cord (Figures 5 and 6). The AAV administration and GFP expression had no untoward effect on motor performance in these rats as studied by rotarod (Supplementary Figure S3). AAV8, AAV9, and AAV10 were compared in adult rats. AAVs were administered into the tail vein, and fluorescence was evaluated 4 weeks post-administration. AAV9 produced greater expression levels of GFP compared to AAV8 and AAV10 (Figures 5 and 6). AAV9 therefore appeared stronger in the adult but not in neonatal administrations. AAV1 was excluded from the analysis in adults due to an insufficiently low titer, although AAV1 GFP was applied to one adult subject. The AAV1 GFP adult subject did not appear to be strongly transduced relative to AAV9 (Figure 5).


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)

GFP expressed by AAV1, AAV8, AAV9, or AAV10 after i.v. vector administration to either neonates or adults. The GFP expression in the CNS appeared to be neuronal with any of the serotypes tested at either age. The different serotypes are shown in the different columns, and the spinal cord and cerebellum are shown at each age. The serotypes performed similarly for GFP expression in the neonates, but in adults, AAV9 appeared to result in the strongest GFP expression. Quantifications of the GFP expression are in Figure 6. Bar in a = 134 μm. Same magnification in b–h. Bar in i = 268 μm. Same magnification in j–l. Bar in m = 536 μm. Same magnification in n–p. CNS, central nervous system; 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

fig5: GFP expressed by AAV1, AAV8, AAV9, or AAV10 after i.v. vector administration to either neonates or adults. The GFP expression in the CNS appeared to be neuronal with any of the serotypes tested at either age. The different serotypes are shown in the different columns, and the spinal cord and cerebellum are shown at each age. The serotypes performed similarly for GFP expression in the neonates, but in adults, AAV9 appeared to result in the strongest GFP expression. Quantifications of the GFP expression are in Figure 6. Bar in a = 134 μm. Same magnification in b–h. Bar in i = 268 μm. Same magnification in j–l. Bar in m = 536 μm. Same magnification in n–p. CNS, central nervous system; GFP, green fluorescent protein; i.v., intravenous.
Mentions: Next, we compared several AAV serotypes that have been used extensively in animals before, such as AAV1, AAV8, AAV9, and AAV10. The same GFP expression plasmid DNA was packaged into each type, and the same dose of viral particles were administered. GFP fluorescence was evaluated in the cerebellum and spinal cord 4 weeks post-administration. With neonatal administrations, we found no significant differences in transduction efficiency in the cerebellum or spinal cord (Figures 5 and 6). The AAV administration and GFP expression had no untoward effect on motor performance in these rats as studied by rotarod (Supplementary Figure S3). AAV8, AAV9, and AAV10 were compared in adult rats. AAVs were administered into the tail vein, and fluorescence was evaluated 4 weeks post-administration. AAV9 produced greater expression levels of GFP compared to AAV8 and AAV10 (Figures 5 and 6). AAV9 therefore appeared stronger in the adult but not in neonatal administrations. AAV1 was excluded from the analysis in adults due to an insufficiently low titer, although AAV1 GFP was applied to one adult subject. The AAV1 GFP adult subject did not appear to be strongly transduced relative to AAV9 (Figure 5).

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