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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.


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Progressive and dose-dependent TDP-43–induced disease state after i.v. administration of AAV9 TDP-43 to adult female rats. (a) The medium-dose of AAV9 TDP-43 (closed squares), but not the low-dose (open squares), reduced weight gain compared to a group of uninjected rats (open circles) on week 3 (*P < 0.05; N = 3–4/group, ANOVA/Bonferroni). (b) The medium-dose AAV9 TDP-43 rats were significantly impaired on the rotarod at week 3 compared to baseline (*P < 0.05, ANOVA/Bonferroni) and (c) also impaired in terms of rearing at week 3 compared to baseline (*P < 0.05, ANOVA/Bonferroni). ANOVA, analysis of variance.
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fig7: Progressive and dose-dependent TDP-43–induced disease state after i.v. administration of AAV9 TDP-43 to adult female rats. (a) The medium-dose of AAV9 TDP-43 (closed squares), but not the low-dose (open squares), reduced weight gain compared to a group of uninjected rats (open circles) on week 3 (*P < 0.05; N = 3–4/group, ANOVA/Bonferroni). (b) The medium-dose AAV9 TDP-43 rats were significantly impaired on the rotarod at week 3 compared to baseline (*P < 0.05, ANOVA/Bonferroni) and (c) also impaired in terms of rearing at week 3 compared to baseline (*P < 0.05, ANOVA/Bonferroni). ANOVA, analysis of variance.

Mentions: There was a clear vector dose dependence of the TDP-43–induced paralysis in adult rats over an 8-week time interval after gene transfer. The low-dose AAV9 TDP-43 group showed no significant differences in weight gain compared to uninjected animals and displayed no signs of motor deficits over 8 weeks (Figure 7). In contrast, the medium dose AAV9 TDP-43 produced a progressive disease state with reduced weight gain, rotarod impairments, and reduced rearing over 3 weeks (Figure 7). The one high dose rat became severely immobilized and died by 3 weeks. Upon evaluating TDP-43 levels in the tissues by western blots, we found that the level of impairment in the low- and medium-dose groups corresponded with TDP-43 expression. Both total and human TDP-43 levels were significantly higher in the medium-dose group than those in the low-dose group (Figure 8; P < 0.05), demonstrating fine dose control of consistent and physiologically relevant TDP-43 levels.


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)

Progressive and dose-dependent TDP-43–induced disease state after i.v. administration of AAV9 TDP-43 to adult female rats. (a) The medium-dose of AAV9 TDP-43 (closed squares), but not the low-dose (open squares), reduced weight gain compared to a group of uninjected rats (open circles) on week 3 (*P < 0.05; N = 3–4/group, ANOVA/Bonferroni). (b) The medium-dose AAV9 TDP-43 rats were significantly impaired on the rotarod at week 3 compared to baseline (*P < 0.05, ANOVA/Bonferroni) and (c) also impaired in terms of rearing at week 3 compared to baseline (*P < 0.05, ANOVA/Bonferroni). ANOVA, analysis of variance.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig7: Progressive and dose-dependent TDP-43–induced disease state after i.v. administration of AAV9 TDP-43 to adult female rats. (a) The medium-dose of AAV9 TDP-43 (closed squares), but not the low-dose (open squares), reduced weight gain compared to a group of uninjected rats (open circles) on week 3 (*P < 0.05; N = 3–4/group, ANOVA/Bonferroni). (b) The medium-dose AAV9 TDP-43 rats were significantly impaired on the rotarod at week 3 compared to baseline (*P < 0.05, ANOVA/Bonferroni) and (c) also impaired in terms of rearing at week 3 compared to baseline (*P < 0.05, ANOVA/Bonferroni). ANOVA, analysis of variance.
Mentions: There was a clear vector dose dependence of the TDP-43–induced paralysis in adult rats over an 8-week time interval after gene transfer. The low-dose AAV9 TDP-43 group showed no significant differences in weight gain compared to uninjected animals and displayed no signs of motor deficits over 8 weeks (Figure 7). In contrast, the medium dose AAV9 TDP-43 produced a progressive disease state with reduced weight gain, rotarod impairments, and reduced rearing over 3 weeks (Figure 7). The one high dose rat became severely immobilized and died by 3 weeks. Upon evaluating TDP-43 levels in the tissues by western blots, we found that the level of impairment in the low- and medium-dose groups corresponded with TDP-43 expression. Both total and human TDP-43 levels were significantly higher in the medium-dose group than those in the low-dose group (Figure 8; P < 0.05), demonstrating fine dose control of consistent and physiologically relevant TDP-43 levels.

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