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Gene therapy in a humanized mouse model of familial hypercholesterolemia leads to marked regression of atherosclerosis.

Kassim SH, Li H, Vandenberghe LH, Hinderer C, Bell P, Marchadier D, Wilson A, Cromley D, Redon V, Yu H, Wilson JM, Rader DJ - PLoS ONE (2010)

Bottom Line: A single intravenous injection of AAV8.mLDLR was found to significantly reduce plasma cholesterol and non-HDL cholesterol levels in chow-fed animals at doses as low as 3×10(9) genome copies/mouse.Collectively, the results presented herein suggest that AAV8-based gene therapy for FH may be feasible and support further development of this approach.The pre-clinical data from these studies will enable for the effective translation of gene therapy into the clinic for treatment of FH.

View Article: PubMed Central - PubMed

Affiliation: Gene Therapy Program, Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.

ABSTRACT

Background: Familial hypercholesterolemia (FH) is an autosomal codominant disorder caused by mutations in the low-density lipoprotein receptor (LDLR) gene. Homozygous FH patients (hoFH) have severe hypercholesterolemia leading to life threatening atherosclerosis in childhood and adolescence. Mice with germ line interruptions in the Ldlr and Apobec1 genes (Ldlr(-/-)Apobec1(-/-)) simulate metabolic and clinical aspects of hoFH, including atherogenesis on a chow diet.

Methods/principal findings: In this study, vectors based on adeno-associated virus 8 (AAV8) were used to deliver the gene for mouse Ldlr (mLDLR) to the livers of Ldlr(-/-)Apobec1(-/-) mice. A single intravenous injection of AAV8.mLDLR was found to significantly reduce plasma cholesterol and non-HDL cholesterol levels in chow-fed animals at doses as low as 3×10(9) genome copies/mouse. Whereas Ldlr(-/-)Apobec1(-/-) mice fed a western-type diet and injected with a control AAV8. vector experienced a further 65% progression in atherosclerosis over 2 months compared with baseline mice, Ldlr(-/-)Apobec1(-/-) mice treated with AAV8.mLDLR realized an 87% regression of atherosclerotic lesions after 3 months compared to baseline mice. Immunohistochemical analyses revealed a substantial remodeling of atherosclerotic lesions.

Conclusions/significance: Collectively, the results presented herein suggest that AAV8-based gene therapy for FH may be feasible and support further development of this approach. The pre-clinical data from these studies will enable for the effective translation of gene therapy into the clinic for treatment of FH.

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Evaluation of AAV8.TBG.mLDLR vector in high fat fed Ldlr-/-Apobec1-/- mice.Amounts of (A) Plasma cholesterol (B) non-HDL cholesterol, and (C) Alanine transaminase were evaluated in Ldlr-/-Apobec1-/- mice up to day 60 after treatment with 1x10∧11 GC of AAV8.TBG.mLDLR (n = 10) or 1×1011 GC of AAV8.TBG.nLacZ (n = 9). Each point represents mean ± s.d. *P<0.05, ‡ P<0.001.
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pone-0013424-g004: Evaluation of AAV8.TBG.mLDLR vector in high fat fed Ldlr-/-Apobec1-/- mice.Amounts of (A) Plasma cholesterol (B) non-HDL cholesterol, and (C) Alanine transaminase were evaluated in Ldlr-/-Apobec1-/- mice up to day 60 after treatment with 1x10∧11 GC of AAV8.TBG.mLDLR (n = 10) or 1×1011 GC of AAV8.TBG.nLacZ (n = 9). Each point represents mean ± s.d. *P<0.05, ‡ P<0.001.

Mentions: Given that AAV8.TBG.mLDLR markedly lowered total cholesterol and non-HDL cholesterol, we next examined whether AAV8.TBG.mLDLR had any effect on the progression or even the regression of atherosclerotic lesions. Three groups of male Ldlr−/−Apobec1−/− mice were fed a high-fat western-type diet to hasten the progression of atherosclerosis. After two months of the atherogenic diet, one group of mice received a single intravenous injection of 1×1011 GC/mouse of control AAV8.TBG.nLacZ vector, one group received 1×1011 GC/mouse of AAV8.TBG.mLDLR vector, while a third baseline group of animals was necropsied for atherosclerosis lesion quantification. The mice who received vectors were maintained on the high-fat diet for an additional 60 days at which time they were necropsied. Animals that received the AAV8.TBG.mLDLR vector realized a rapid drop in total cholesterol from 1555±343 mg/dl at baseline to 266±78 at day 7 and to 67±13 by day 60 after treatment (Fig. 4A). By contrast, the plasma cholesterol levels of AAV8.TBG.nLacZ treated mice remained virtually unchanged from 1566±276 at baseline to 1527±67 when measured 60 days after vector (Fig. 4A). The same trend was observed with respect to non-HDL cholesterol (Fig. 4B). All animals developed slight increases in serum transaminanses following the 2 months on the high fat diet, which remained elevated following treatment with the AAV8.TBG.nLacZ vector but diminished three-fold to normal levels after treatment with the AAV8.TBG.mLDLR vector (Fig. 4C).


Gene therapy in a humanized mouse model of familial hypercholesterolemia leads to marked regression of atherosclerosis.

Kassim SH, Li H, Vandenberghe LH, Hinderer C, Bell P, Marchadier D, Wilson A, Cromley D, Redon V, Yu H, Wilson JM, Rader DJ - PLoS ONE (2010)

Evaluation of AAV8.TBG.mLDLR vector in high fat fed Ldlr-/-Apobec1-/- mice.Amounts of (A) Plasma cholesterol (B) non-HDL cholesterol, and (C) Alanine transaminase were evaluated in Ldlr-/-Apobec1-/- mice up to day 60 after treatment with 1x10∧11 GC of AAV8.TBG.mLDLR (n = 10) or 1×1011 GC of AAV8.TBG.nLacZ (n = 9). Each point represents mean ± s.d. *P<0.05, ‡ P<0.001.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2957433&req=5

pone-0013424-g004: Evaluation of AAV8.TBG.mLDLR vector in high fat fed Ldlr-/-Apobec1-/- mice.Amounts of (A) Plasma cholesterol (B) non-HDL cholesterol, and (C) Alanine transaminase were evaluated in Ldlr-/-Apobec1-/- mice up to day 60 after treatment with 1x10∧11 GC of AAV8.TBG.mLDLR (n = 10) or 1×1011 GC of AAV8.TBG.nLacZ (n = 9). Each point represents mean ± s.d. *P<0.05, ‡ P<0.001.
Mentions: Given that AAV8.TBG.mLDLR markedly lowered total cholesterol and non-HDL cholesterol, we next examined whether AAV8.TBG.mLDLR had any effect on the progression or even the regression of atherosclerotic lesions. Three groups of male Ldlr−/−Apobec1−/− mice were fed a high-fat western-type diet to hasten the progression of atherosclerosis. After two months of the atherogenic diet, one group of mice received a single intravenous injection of 1×1011 GC/mouse of control AAV8.TBG.nLacZ vector, one group received 1×1011 GC/mouse of AAV8.TBG.mLDLR vector, while a third baseline group of animals was necropsied for atherosclerosis lesion quantification. The mice who received vectors were maintained on the high-fat diet for an additional 60 days at which time they were necropsied. Animals that received the AAV8.TBG.mLDLR vector realized a rapid drop in total cholesterol from 1555±343 mg/dl at baseline to 266±78 at day 7 and to 67±13 by day 60 after treatment (Fig. 4A). By contrast, the plasma cholesterol levels of AAV8.TBG.nLacZ treated mice remained virtually unchanged from 1566±276 at baseline to 1527±67 when measured 60 days after vector (Fig. 4A). The same trend was observed with respect to non-HDL cholesterol (Fig. 4B). All animals developed slight increases in serum transaminanses following the 2 months on the high fat diet, which remained elevated following treatment with the AAV8.TBG.nLacZ vector but diminished three-fold to normal levels after treatment with the AAV8.TBG.mLDLR vector (Fig. 4C).

Bottom Line: A single intravenous injection of AAV8.mLDLR was found to significantly reduce plasma cholesterol and non-HDL cholesterol levels in chow-fed animals at doses as low as 3×10(9) genome copies/mouse.Collectively, the results presented herein suggest that AAV8-based gene therapy for FH may be feasible and support further development of this approach.The pre-clinical data from these studies will enable for the effective translation of gene therapy into the clinic for treatment of FH.

View Article: PubMed Central - PubMed

Affiliation: Gene Therapy Program, Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.

ABSTRACT

Background: Familial hypercholesterolemia (FH) is an autosomal codominant disorder caused by mutations in the low-density lipoprotein receptor (LDLR) gene. Homozygous FH patients (hoFH) have severe hypercholesterolemia leading to life threatening atherosclerosis in childhood and adolescence. Mice with germ line interruptions in the Ldlr and Apobec1 genes (Ldlr(-/-)Apobec1(-/-)) simulate metabolic and clinical aspects of hoFH, including atherogenesis on a chow diet.

Methods/principal findings: In this study, vectors based on adeno-associated virus 8 (AAV8) were used to deliver the gene for mouse Ldlr (mLDLR) to the livers of Ldlr(-/-)Apobec1(-/-) mice. A single intravenous injection of AAV8.mLDLR was found to significantly reduce plasma cholesterol and non-HDL cholesterol levels in chow-fed animals at doses as low as 3×10(9) genome copies/mouse. Whereas Ldlr(-/-)Apobec1(-/-) mice fed a western-type diet and injected with a control AAV8. vector experienced a further 65% progression in atherosclerosis over 2 months compared with baseline mice, Ldlr(-/-)Apobec1(-/-) mice treated with AAV8.mLDLR realized an 87% regression of atherosclerotic lesions after 3 months compared to baseline mice. Immunohistochemical analyses revealed a substantial remodeling of atherosclerotic lesions.

Conclusions/significance: Collectively, the results presented herein suggest that AAV8-based gene therapy for FH may be feasible and support further development of this approach. The pre-clinical data from these studies will enable for the effective translation of gene therapy into the clinic for treatment of FH.

Show MeSH
Related in: MedlinePlus