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Molybdenum cofactor deficiency type B knock-in mouse models carrying patient-identical mutations and their rescue by singular AAV injections

Abstract

Molybdenum cofactor deficiency is an autosomal, recessively inherited metabolic disorder, which, in the absence of an effective therapy, leads to early childhood death due to neurological deterioration. In type A of the disease, cyclic pyranopterin monophosphate (cPMP) is missing, the first intermediate in the biosynthesis of the cofactor, and a biochemical substitution therapy using cPMP has been developed. A comparable approach for type B of the disease with a defect in the second step of the synthesis, formation of molybdopterin, so far has been hampered by the extreme instability of the corresponding metabolites. To explore avenues for a successful and safe gene therapy, knock-in mouse models were created carrying the mutations c.88C>T (p.Q30X) and c.726_727delAA, which are also found in human patients. Recombinant adeno-associated viruses (rAAVs) were constructed and used for postnatal intrahepatic injections of MoCo-deficient mice in a proof-of-concept approach. Singular administration of an appropriate virus dose in 60 animals prevented the otherwise devastating phenotype to a variable extent. While untreated mice did not survive for more than 2 weeks, some of the treated mice grew up to adulthood in both sexes.

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Acknowledgements

This study was supported by the Deutsche Forschungsgemeinschaft (DFG RE768/18-1). Construction of knock-in mice and gene therapy vectors was performed by commercial entities as indicated in the text. Intellectual property rights are owned by the author, the Universitätsmedizin Göttingen and the Deutsche Forschungsgemeinschaft. I thank Christina Ahlbrecht for invaluable help with the animals and laboratory work.

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Correspondence to Jochen Reiss.

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The author declares no conflict of interest.

Statement of human/animal rights

This article does not contain any studies with human participants.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted (Nds. Landesamt für Verbraucherschutz und Lebensmittelsicherheit (LAVES), Dezernat 33, Röverskamp 5, 26203 Wardenburg).

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Supplementary material 1 Untreated homozygous Mocs2A-deficient mouse (right) on day 8 after birth in comparison to heterozygous littermate (left) (AVI 28054 KB)

Supplementary material 1 Untreated homozygous Mocs2A-deficient mouse (right) on day 8 after birth in comparison to heterozygous littermate (left) (AVI 28054 KB)

Supplementary material 2 Untreated homozygous Mocs2B-deficient mouse (right) on day 6 after birth in comparison to heterozygous littermate (left) (AVI 6835 KB)

Supplementary material 2 Untreated homozygous Mocs2B-deficient mouse (right) on day 6 after birth in comparison to heterozygous littermate (left) (AVI 6835 KB)

Supplementary material 3 Homozygous Mocs2A-deficient mouse injected at day 4 after birth (left) at age of 4 months in comparison to age-matched control (right). The speed of the rotating rod is 20 rpm (AVI 65080 KB)

Supplementary material 3 Homozygous Mocs2A-deficient mouse injected at day 4 after birth (left) at age of 4 months in comparison to age-matched control (right). The speed of the rotating rod is 20 rpm (AVI 65080 KB)

Supplementary material 4 Homozygous Mocs2B-deficient mouse injected at day 2 after birth on day 14 after birth (AVI 27926 KB)

Supplementary material 4 Homozygous Mocs2B-deficient mouse injected at day 2 after birth on day 14 after birth (AVI 27926 KB)

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Reiss, J. Molybdenum cofactor deficiency type B knock-in mouse models carrying patient-identical mutations and their rescue by singular AAV injections. Hum Genet 138, 355–361 (2019) doi:10.1007/s00439-019-01992-z

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