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Adeno-associated viral vectors for the treatment of glycogen storage diseases

A carrier and virus technology, applied in the direction of virus/bacteriophage, virus, vector, etc., can solve the problem of not being able to completely correct liver G6Pase-α deficiency

Active Publication Date: 2020-08-04
UNITED STATES OF AMERICA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although these studies are promising, none of them have been able to completely correct the hepatic G6Pase-α deficiency.

Method used

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  • Adeno-associated viral vectors for the treatment of glycogen storage diseases
  • Adeno-associated viral vectors for the treatment of glycogen storage diseases
  • Adeno-associated viral vectors for the treatment of glycogen storage diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Example 1: Complete normalization of hepatic G6PC deficiency in glycogen storage disease type Ia using gene therapy

[0132] This example describes a comparison of the hepatic gene delivery efficiency and G6Pase-α expression of two G6Pase-α expressing AAV vectors driven by two different promoters in G6PC deficient mice. Results demonstrate that an AAV vector with a G6PC promoter / enhancer (AAV-GPE) is more efficient at directing durable in vivo hepatic transgene expression than an AAV vector with a chicken β-actin promoter / CMV enhancer (AAV-CBA) . In addition, G6PC-deficient mice treated with AAV-GPE exhibited normal levels of blood glucose, blood metabolites, liver glycogen, and liver fat.

[0133] Materials and methods

[0134] Construction of pUF11-GPE-G6PC and preparation of AAV vector

[0135] pUF11-mG6Pase-α-CBA (Ghosh et al., Gene Ther13:321-329, 2006) (wherein, mouse G6Pase-α is driven by CBA promoter / CMV enhancer (Xu et al., Hum Gene Ther 12:563-573, 2001)) ...

Embodiment 2

[0186] Example 2: Prevention of hepatocellular adenoma and correction of metabolic abnormalities in type IA glycogen storage disease using gene therapy

[0187] This example describes studies evaluating the efficacy of AAV-GPE vectors in long-term studies. G6pc - / - AAV-GPE vector-mediated gene therapy in mice was effective for at least 70-90 weeks in mice expressing greater than 3% of wild-type hepatic G6Pase-α. The results demonstrated that AAV-GPE-treated mice exhibited normal hepatic fat storage, normal blood metabolites and glucose tolerance profiles, reduced fasting blood insulin levels, and no signs of liver abnormalities.

[0188] Materials and methods

[0189] To G6pc - / - Mice infused with AAV-GPE

[0190] AAV-GPE vector (as described in Example 1, Yiu et al., Mol Ther 18:1076-1084, 2010) was infused into G6pc via the retro-orbital sinus - / - in mice (Lei et al., Nat Genet 13:203-209, 1996). Use an age-matched G6pc + / + / G6pc + / - and 6- to 10-week-old G6pc - / - M...

Embodiment 3

[0225] Example 3: The upstream enhancer element of the G6PC promoter is optimal for type Ia glycogen storage disease

[0226] G6PC expression is critical

[0227]The studies described in this example further evaluated the efficacy of AAV-GPE vectors. AAV-GPE is a single-stranded vector containing a G6PC promoter / enhancer of 2684bp compared to the double-stranded vector AAV-miGPE containing a minimal G6PC promoter / enhancer of 382bp. The results described in this example demonstrate that AAV-GPE vectors direct significantly higher levels of hepatic G6Pase-α expression in GSD-Ia mice compared to AAV-miGPE vectors, resulting in greater hepatic glycogen accumulation. and better fasting tolerance.

[0228] Materials and methods

[0229] Infusion of rAAV vectors into G6pc- / - mice

[0230] All G6pc- / - mice were kept alive by glucose therapy (Lei et al., Nat. Genet. 13:203-209, 1996). The rAAV vector was infused into 2-week-old G6pc- / - mice via the retro-orbital sinus. Use an age...

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Abstract

The present disclosure describes improved adeno-associated virus (AAV) vectors for gene therapy applications in the treatment of glycogen storage diseases, particularly type Ia glycogen storage diseases. Recombinant nucleic acid molecules, vectors, and recombinant AAV comprising a G6PC promoter / enhancer, a synthetic intron, a G6PC coding sequence (such as a wild-type or codon-optimized G6PC coding sequence), and and the stuffer nucleic acid sequences between the intron and the G6PC coding sequence. The recombinant AAVs disclosed herein exhibit efficient liver transduction and are able to correct metabolic abnormalities in animal models of GSD-Ia.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Application No. 61 / 908,861, filed November 26, 2013, which is incorporated herein by reference in its entirety. technical field [0003] The present disclosure relates to gene therapy vectors for the treatment of glycogen storage diseases, particularly type Ia glycogen storage diseases. Background technique [0004] Glycogen storage disease type Ia (GSD-Ia or von Gierke disease, MIM232200) is caused by a defect in glucose-6-phosphatase alpha (G6Pase-α), a , an enzyme expressed in the kidney and intestine (Chou et al., Nat Rev Endocrinol 6:676-688, 2010). G6Pase-α, encoded by the G6PC gene, is a hydrophobic protein anchored in the endoplasmic reticulum (ER) by nine transmembrane helices (Chou et al., Nat Rev Endocrinol 6:676-688, 2010). This enzyme catalyzes the hydrolysis of glucose-6-phosphate (G6P) to glucose and phosphate in the final step of glycogenolysis and gl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/16A61K48/00C12N15/86
CPCA61K48/005C12N9/16C12Y301/03009C12N2750/14143C12N2830/008C12N2830/42C12N15/86A61K48/0066A61K48/0075A61P1/16A61P13/12A61P19/06A61P3/06A61P3/08A61P3/10C12N7/00A61K48/0008
Inventor J·J·周B·J·伯恩
Owner UNITED STATES OF AMERICA