Liver-Directed Recombinant Adeno-Associated Viral Gene Delivery Rescues a Lethal Mouse Model of Methylmalonic Acidemia and Provides Long-Term Phenotypic Correction

Organic Acid Research Section, Genetics and Molecular Biology Branch, National Human Genome Research Institute, National Institutes of Health Bethesda, MD 20892, USA.
Human gene therapy (Impact Factor: 3.76). 09/2010; 21(9):1147-54. DOI: 10.1089/hum.2010.008
Source: PubMed


Methylmalonic acidemia is a severe metabolic disorder caused by a deficiency of the ubiquitously expressed mitochondrial enzyme, methylmalonyl-CoA mutase (MUT). Liver transplantation has been used to treat a small number of patients with variable success, and whether liver-directed gene therapy might be employed in such a pleiotropic metabolic disorder is uncertain. In this study, we examined the therapeutic effects of hepatocyte-directed delivery of the Mut gene to mice with a severe form of methylmalonic acidemia. We show that a single intrahepatic injection of recombinant adeno-associated virus serotype 8 expressing the Mut gene under the control of the liver-specific thyroxine-binding globulin (TBG) promoter is sufficient to rescue Mut(-/-) mice from neonatal lethality and provide long-term phenotypic correction. Treated Mut(-/-) mice lived beyond 1 year of age, had improved growth, lower plasma methylmalonic acid levels, and an increased capacity to oxidize [1-(13)C]propionate in vivo. The older treated mice showed increased Mut transcription, presumably mediated by upregulation of the TBG promoter during senescence. The results indicate that the stable transduction of a small number of hepatocytes with the Mut gene can be efficacious in the phenotypic correction of an inborn error of organic acid metabolism and support the rapid translation of liver-directed gene therapy vectors already optimized for human subjects to patients with methylmalonic acidemia.

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Available from: Randy J. Chandler, Oct 07, 2015
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    • "The use of chemical-based carriers, such as cell-penetrating peptides for oligonucleotide delivery41 or encapsulated liposomes42 may also obviate the need for hydrodynamic delivery of the AmiR. Furthermore, the technique is not limited to plasmids, since it should also be possible to use viral vectors with hepatic tropism for the delivery.43,44 "
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    ABSTRACT: Inducible systems for gene expression emerge as a new class of artificial vectors offering temporal and spatial exogenous control of gene expression. However, most inducible systems are less efficient in vivo and lack the target-organ specificity. In the present study, we have developed and optimized an oligonucleotide-based inducible system for the in vivo control of transgenes in the liver. We generated a set of simple, inducible plasmid-vectors based on the addition of four units of liver-specific miR-122 target sites to the 3'untranslated region of the gene of interest. Once the vector was delivered into hepatocytes this modification induced a dramatic reduction of gene expression that could be restored by the infusion of an antagomir for miR-122. The efficiency of the system was tested in vivo, and displayed low background and strong increase in gene expression upon induction. Moreover, gene expression was repeatedly induced even several months after the first induction showing no toxic effect in vivo. By combining tissue-specific control elements with antagomir treatment we generated, optimized and validated a robust inducible system that could be used successfully for in vivo experimental models requiring tight and cyclic control of gene expression.
    Molecular Therapy - Nucleic Acids 07/2014; 3(7):e172. DOI:10.1038/mtna.2014.25 · 4.51 Impact Factor
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    • "Neither the elimination of transgene expression nor the impaired mitochondrial importation and processing of the human MUT protein is suggested by our studies. The fact that circulating metabolite concentrations and in vivo propionate oxidative capacity in the AAV treated Mut −/− mice is almost exactly what we previously observed in studies using AAVs that expressed the murine cDNA provide support for this claim [5] [6]. While the presence of antibodies against MUT was not directly investigated, the prolonged survival after treatment suggests the absence of an immune response. "
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    ABSTRACT: We demonstrate that human methylmalonyl-CoA mutase (MUT), delivered using an AAV serotype 8 vector, rescues the lethal phenotype displayed by mice with MMA and provides long-term phenotypic correction. In addition to defining a lower limit of effective dosing, our studies establish that neither a species barrier to mitochondrial processing nor an apparent immune response to MUT limits the murine model as an experimental platform to test the efficacy of human gene therapy vectors for MMA. Published by Elsevier Inc.
    Molecular Genetics and Metabolism 09/2012; 107(3). DOI:10.1016/j.ymgme.2012.09.019 · 2.63 Impact Factor
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    • "In this study they characterised the model using many more uninjected mice (58 versus 17 previously). They demonstrated much more effective correction with 5 out of 6 mice which had received 4x1011 AAV genomes surviving for more than one year [103]. The improvement in survival compared with the adenoviral vector administered mice may be attributable to both the increased efficiency of transduction of the AAV vector and also the resistance to silencing of the thyroxine-binding globulin promoter compared with the CMV promoter. "
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    ABSTRACT: The liver acts as a host to many functions hence raising the possibility that any one may be compromised by a single gene defect. Inherited or de novo mutations in these genes may result in relatively mild diseases or be so devastating that death within the first weeks or months of life is inevitable. Some diseases can be managed using conventional medicines whereas others are, as yet, untreatable. In this review we consider the application of early intervention gene therapy in neonatal and fetal preclinical studies. We appraise the tools of this technology, including lentivirus, adenovirus and adeno-associated virus (AAV)-based vectors. We highlight the application of these for a range of diseases including hemophilia, urea cycle disorders such as ornithine transcarbamylase deficiency, organic acidemias, lysosomal storage diseases including mucopolysaccharidoses, glycogen storage diseases and bile metabolism. We conclude by assessing the advantages and disadvantages associated with fetal and neonatal liver gene transfer.
    Current pharmaceutical design 07/2011; 17(24):2528-41. DOI:10.2174/138161211797247541 · 3.45 Impact Factor
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