Searchable abstracts of presentations at key conferences in endocrinology
Endocrine Abstracts (2009) 19 OC31

SFEBES2009 Oral Communications Bone and Calcium (8 abstracts)

A mouse with a Trp589Arg mutation in N-acetylgalactosaminyltransferase 3 (Galnt3) provides a model for familial tumoural calcinosis

C Esapa 1, , R Head 1, , C Chan 1, , E Crane 1, , M Cheeseman 2 , T Hough 2 , E McNally 3 , A Carr 4 , G Thomas 5 , M Brown 5 , P Croucher 6 , S Brown 2 , R Cox 2 & R Thakker 1

1Nuffield Department of Clinical Medicine, University of Oxford, Oxford, UK; 2Harwell Science and Innovation Campus, MRC Harwell, Harwell, UK; 3Department of Radiology, Nuffield Orthopaedic Centre and Nuffield Department of Orthopaedic Surgery, University of Oxford, Oxford, UK; 4NIHR Biomedical Research Unit, Nuffield Department of Orthopaedic Surgery, University of Oxford, Oxford, UK; 5University of Queensland, Queensland, Australia; 6University of Sheffield, Sheffield, UK.

Investigations of bone disorders which are often inherited have yielded important insights in the molecular mechanisms of bone development, osteoporosis and osteoarthritis. However, these studies have been hampered by the lack of available patients and affected families. To overcome this limitation, we have investigated mice treated with the chemical mutagen N-ethyl-N-nitrosourea (ENU) for hereditary bone disorders. Mice were kept in accordance with national welfare guidelines and project license restrictions. At 12 weeks of age, all mice underwent phenotypic assessments that included dysmorphology, radiography and serum biochemistry. These processes identified a mouse mutant that presented with hyperphosphataemia and progressive ectopic calcification. This phenotype was inherited in an autosomal recessive manner. Histological analysis revealed calcification in multiple tissues that included striated muscle, kidney and testis. These features are consistent with familial tumoural calcinosis (FTC) which in man is caused by mutations in fibroblast growth factor 23 (FGF23), KLOTHO, or N-acetylgalactosaminyltransferase 3 (GALNT3). DNA sequence analysis of the mouse homologous genes identified a T to A transversion at nucleotide 1765 of the Galnt3 gene. This altered a highly conserved tryptophan (Trp) residue at position 589 to an arginine (Arg) residue. The presence of this missense Trp589Arg mutation was confirmed using the amplification refractory mutation system (ARMS) PCR technique. ARMS-PCR was performed on genomic DNA using two primer pairs to respectively amplify the wild type and mutant alleles, followed by agarose gel electrophoresis. Heterozygous (+/−) mice were phenotypically normal whilst homozygous (−/−) mice had hyperphosphataemia in association with calcinosis. Thus, we have established a mouse model for autosomal recessive FTC. These results will help to increase our understanding of the molecular mechanisms of FTC and provide a useful model for investigating novel treatments.

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