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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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Wide-scale neuronal transduction in the CNS and other tissues after AAV9 GFP tail vein injections to adult rats. Transduced neurons were seen in all areas of the CNS that were examined. Some of the transduced regions included (a, b) hippocampus and thalamus, (c, d) cortex, (e, f) striatum (note morphology of medium spiny neurons), (g) substantia nigra, and (h) dorsal root ganglia (DRG). Outside of the CNS, there was substantial transduction of (i) the liver, and (j) the heart. Sparse GFP-labeling was seen in the (k) kidney and the (l) lung. Bars in a and e = 536 μm. Bars in b, c, g, and i = 268 μm. Bars in d, f, h, and j–l = 134 μm. CNS, central nervous system; GFP, green fluorescent protein.
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fig2: Wide-scale neuronal transduction in the CNS and other tissues after AAV9 GFP tail vein injections to adult rats. Transduced neurons were seen in all areas of the CNS that were examined. Some of the transduced regions included (a, b) hippocampus and thalamus, (c, d) cortex, (e, f) striatum (note morphology of medium spiny neurons), (g) substantia nigra, and (h) dorsal root ganglia (DRG). Outside of the CNS, there was substantial transduction of (i) the liver, and (j) the heart. Sparse GFP-labeling was seen in the (k) kidney and the (l) lung. Bars in a and e = 536 μm. Bars in b, c, g, and i = 268 μm. Bars in d, f, h, and j–l = 134 μm. CNS, central nervous system; GFP, green fluorescent protein.

Mentions: AAV9 GFP at a dose of 1 × 1014 vector genomes (vg)/kg produced wide-scale neuronal transduction throughout the spinal cord and the cerebellum (Figure 1 and Supplementary Figure S1), two areas that are also highly transduced with the neonatal approach. Transduction of spinal motor neurons and cerebellar Purkinje neurons are shown in Figure 1. We counterstained adjacent sections with an astroglial marker and observed that very few if any of the transduced cells were astrocytes in the adult rats. Thus, the same transduction pattern of AAV9 within the CNS resulted after tail vein injections to adults as occurs after either i.v. administration to neonatal rats or with stereotaxic injections (Figures 1 and 2). Viewing the Purkinje cell layer in the cerebellum of adult rats with robust expression in this area, we did not notice signs of cellular infiltration relative to controls by staining with hematoxylin and eosin (Supplementary Figure S2). Outside of the CNS, with this promoter system and i.v. administration in either neonates or adults, there is significant expression in several peripheral organs, such as the liver and the heart (Figure 2).


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)

Wide-scale neuronal transduction in the CNS and other tissues after AAV9 GFP tail vein injections to adult rats. Transduced neurons were seen in all areas of the CNS that were examined. Some of the transduced regions included (a, b) hippocampus and thalamus, (c, d) cortex, (e, f) striatum (note morphology of medium spiny neurons), (g) substantia nigra, and (h) dorsal root ganglia (DRG). Outside of the CNS, there was substantial transduction of (i) the liver, and (j) the heart. Sparse GFP-labeling was seen in the (k) kidney and the (l) lung. Bars in a and e = 536 μm. Bars in b, c, g, and i = 268 μm. Bars in d, f, h, and j–l = 134 μm. CNS, central nervous system; GFP, green fluorescent protein.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig2: Wide-scale neuronal transduction in the CNS and other tissues after AAV9 GFP tail vein injections to adult rats. Transduced neurons were seen in all areas of the CNS that were examined. Some of the transduced regions included (a, b) hippocampus and thalamus, (c, d) cortex, (e, f) striatum (note morphology of medium spiny neurons), (g) substantia nigra, and (h) dorsal root ganglia (DRG). Outside of the CNS, there was substantial transduction of (i) the liver, and (j) the heart. Sparse GFP-labeling was seen in the (k) kidney and the (l) lung. Bars in a and e = 536 μm. Bars in b, c, g, and i = 268 μm. Bars in d, f, h, and j–l = 134 μm. CNS, central nervous system; GFP, green fluorescent protein.
Mentions: AAV9 GFP at a dose of 1 × 1014 vector genomes (vg)/kg produced wide-scale neuronal transduction throughout the spinal cord and the cerebellum (Figure 1 and Supplementary Figure S1), two areas that are also highly transduced with the neonatal approach. Transduction of spinal motor neurons and cerebellar Purkinje neurons are shown in Figure 1. We counterstained adjacent sections with an astroglial marker and observed that very few if any of the transduced cells were astrocytes in the adult rats. Thus, the same transduction pattern of AAV9 within the CNS resulted after tail vein injections to adults as occurs after either i.v. administration to neonatal rats or with stereotaxic injections (Figures 1 and 2). Viewing the Purkinje cell layer in the cerebellum of adult rats with robust expression in this area, we did not notice signs of cellular infiltration relative to controls by staining with hematoxylin and eosin (Supplementary Figure S2). Outside of the CNS, with this promoter system and i.v. administration in either neonates or adults, there is significant expression in several peripheral organs, such as the liver and the heart (Figure 2).

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