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Assessment of toxicity and biodistribution of recombinant AAV8 vector-mediated immunomodulatory gene therapy in mice with Pompe disease.

Wang G, Young SP, Bali D, Hutt J, Li S, Benson J, Koeberl DD - Mol Ther Methods Clin Dev (2014)

Bottom Line: The vector genome was sustained in all tissues through 16-week postdosing, except for in blood with a similar tissue tropism between males and females.Administration of the vector alone, or combined with the ERT, was effective in producing significantly increased GAA activity and consequently decreased glycogen accumulation in multiple tissues, and the urine biomarker, Glc4, was significantly reduced.The efficacy of the vector (or with ERT) was better in males than in females, as demonstrated both by the number of tissues showing significantly effective responses and the extent of response in a given tissue.

View Article: PubMed Central - PubMed

Affiliation: Applied Toxicology and Gene Therapy Pharm/Tox Program, Lovelace Respiratory Research Institute , Albuquerque, New Mexico, USA.

ABSTRACT
A preclinical safety study was conducted to evaluate the short- and long-term toxicity of a recombinant adeno-associated virus serotype 8 (AAV2/8) vector that has been developed as an immune-modulatory adjunctive therapy to recombinant human acid α-glucosidase (rhGAA, Myozyme) enzyme replacement treatment (ERT) for patients with Pompe disease (AAV2/8-LSPhGAApA). The AAV2/8-LSPhGAApA vector at 1.6 × 10(13) vector particles/kg, after intravenous injection, did not cause significant short- or long-term toxicity. Recruitment of CD4(+) (but not CD8(+)) lymphocytes to the liver was elevated in the vector-dosed male animals at study day (SD) 15, and in group 8 animals at SD 113, in comparison to their respective control animals. Administration of the vector, either prior to or after the one ERT injection, uniformly prevented the hypersensitivity induced by subsequent ERT in males, but not always in female animals. The vector genome was sustained in all tissues through 16-week postdosing, except for in blood with a similar tissue tropism between males and females. Administration of the vector alone, or combined with the ERT, was effective in producing significantly increased GAA activity and consequently decreased glycogen accumulation in multiple tissues, and the urine biomarker, Glc4, was significantly reduced. The efficacy of the vector (or with ERT) was better in males than in females, as demonstrated both by the number of tissues showing significantly effective responses and the extent of response in a given tissue. Given the lack of toxicity for AAV2/8LSPhGAApA, further consideration of clinical translation is warranted in Pompe disease.

No MeSH data available.


Related in: MedlinePlus

Biodistribution of AAV2/8-LSPhGAApA vector in mouse tissues. Tissues were collected from mice administered AAV2/8-LSPhGAApA vector at 1.6 × 1013 vp/kg via tail vein injection at 2, 8, and 16 weeks postdosing. The vector genome copies were analyzed by quantitative polymerase chain reaction (qPCR) as copies/µg genomic DNA. The lowest limit of quantitation of the qPCR assay was 38 copies/µg genomic DNA. The data were presented as geometric mean (n = 5). The positive error bar represents the upper limit of 95% confidence interval, and the negative error bar the lower limit of 95% confidence interval. M tissue was from male mice; F tissue was from female mice. Note that only data from vector-dosed groups (group 10 for 2 weeks, group 12 for 8 weeks, and group 14 for 16 weeks) were presented, and vector genome copy numbers in the vehicle control animal tissues from group 9, 11, and 13 were all below the lower limit of quantification of the qPCR assay (data not shown).
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fig7: Biodistribution of AAV2/8-LSPhGAApA vector in mouse tissues. Tissues were collected from mice administered AAV2/8-LSPhGAApA vector at 1.6 × 1013 vp/kg via tail vein injection at 2, 8, and 16 weeks postdosing. The vector genome copies were analyzed by quantitative polymerase chain reaction (qPCR) as copies/µg genomic DNA. The lowest limit of quantitation of the qPCR assay was 38 copies/µg genomic DNA. The data were presented as geometric mean (n = 5). The positive error bar represents the upper limit of 95% confidence interval, and the negative error bar the lower limit of 95% confidence interval. M tissue was from male mice; F tissue was from female mice. Note that only data from vector-dosed groups (group 10 for 2 weeks, group 12 for 8 weeks, and group 14 for 16 weeks) were presented, and vector genome copy numbers in the vehicle control animal tissues from group 9, 11, and 13 were all below the lower limit of quantification of the qPCR assay (data not shown).

Mentions: The AAV2/8-LSPhGAApA vector, after tail vein injection, was detected at 2 weeks in all the main tissues and blood examined (Figure 7) with a highest level in the liver (~107 vg/μg DNA) and lowest in the brain (~102 vg/μg DNA). The vector persisted in all the examined tissues up to 16 weeks without significant reduction except that in blood the vector level was decreased to around unquantifiable level (the low limit of quantification for the assay is 38 vg/μg DNA). The vector distribution pattern was similar between male and female animals (data not shown).


Assessment of toxicity and biodistribution of recombinant AAV8 vector-mediated immunomodulatory gene therapy in mice with Pompe disease.

Wang G, Young SP, Bali D, Hutt J, Li S, Benson J, Koeberl DD - Mol Ther Methods Clin Dev (2014)

Biodistribution of AAV2/8-LSPhGAApA vector in mouse tissues. Tissues were collected from mice administered AAV2/8-LSPhGAApA vector at 1.6 × 1013 vp/kg via tail vein injection at 2, 8, and 16 weeks postdosing. The vector genome copies were analyzed by quantitative polymerase chain reaction (qPCR) as copies/µg genomic DNA. The lowest limit of quantitation of the qPCR assay was 38 copies/µg genomic DNA. The data were presented as geometric mean (n = 5). The positive error bar represents the upper limit of 95% confidence interval, and the negative error bar the lower limit of 95% confidence interval. M tissue was from male mice; F tissue was from female mice. Note that only data from vector-dosed groups (group 10 for 2 weeks, group 12 for 8 weeks, and group 14 for 16 weeks) were presented, and vector genome copy numbers in the vehicle control animal tissues from group 9, 11, and 13 were all below the lower limit of quantification of the qPCR assay (data not shown).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4362383&req=5

fig7: Biodistribution of AAV2/8-LSPhGAApA vector in mouse tissues. Tissues were collected from mice administered AAV2/8-LSPhGAApA vector at 1.6 × 1013 vp/kg via tail vein injection at 2, 8, and 16 weeks postdosing. The vector genome copies were analyzed by quantitative polymerase chain reaction (qPCR) as copies/µg genomic DNA. The lowest limit of quantitation of the qPCR assay was 38 copies/µg genomic DNA. The data were presented as geometric mean (n = 5). The positive error bar represents the upper limit of 95% confidence interval, and the negative error bar the lower limit of 95% confidence interval. M tissue was from male mice; F tissue was from female mice. Note that only data from vector-dosed groups (group 10 for 2 weeks, group 12 for 8 weeks, and group 14 for 16 weeks) were presented, and vector genome copy numbers in the vehicle control animal tissues from group 9, 11, and 13 were all below the lower limit of quantification of the qPCR assay (data not shown).
Mentions: The AAV2/8-LSPhGAApA vector, after tail vein injection, was detected at 2 weeks in all the main tissues and blood examined (Figure 7) with a highest level in the liver (~107 vg/μg DNA) and lowest in the brain (~102 vg/μg DNA). The vector persisted in all the examined tissues up to 16 weeks without significant reduction except that in blood the vector level was decreased to around unquantifiable level (the low limit of quantification for the assay is 38 vg/μg DNA). The vector distribution pattern was similar between male and female animals (data not shown).

Bottom Line: The vector genome was sustained in all tissues through 16-week postdosing, except for in blood with a similar tissue tropism between males and females.Administration of the vector alone, or combined with the ERT, was effective in producing significantly increased GAA activity and consequently decreased glycogen accumulation in multiple tissues, and the urine biomarker, Glc4, was significantly reduced.The efficacy of the vector (or with ERT) was better in males than in females, as demonstrated both by the number of tissues showing significantly effective responses and the extent of response in a given tissue.

View Article: PubMed Central - PubMed

Affiliation: Applied Toxicology and Gene Therapy Pharm/Tox Program, Lovelace Respiratory Research Institute , Albuquerque, New Mexico, USA.

ABSTRACT
A preclinical safety study was conducted to evaluate the short- and long-term toxicity of a recombinant adeno-associated virus serotype 8 (AAV2/8) vector that has been developed as an immune-modulatory adjunctive therapy to recombinant human acid α-glucosidase (rhGAA, Myozyme) enzyme replacement treatment (ERT) for patients with Pompe disease (AAV2/8-LSPhGAApA). The AAV2/8-LSPhGAApA vector at 1.6 × 10(13) vector particles/kg, after intravenous injection, did not cause significant short- or long-term toxicity. Recruitment of CD4(+) (but not CD8(+)) lymphocytes to the liver was elevated in the vector-dosed male animals at study day (SD) 15, and in group 8 animals at SD 113, in comparison to their respective control animals. Administration of the vector, either prior to or after the one ERT injection, uniformly prevented the hypersensitivity induced by subsequent ERT in males, but not always in female animals. The vector genome was sustained in all tissues through 16-week postdosing, except for in blood with a similar tissue tropism between males and females. Administration of the vector alone, or combined with the ERT, was effective in producing significantly increased GAA activity and consequently decreased glycogen accumulation in multiple tissues, and the urine biomarker, Glc4, was significantly reduced. The efficacy of the vector (or with ERT) was better in males than in females, as demonstrated both by the number of tissues showing significantly effective responses and the extent of response in a given tissue. Given the lack of toxicity for AAV2/8LSPhGAApA, further consideration of clinical translation is warranted in Pompe disease.

No MeSH data available.


Related in: MedlinePlus