title: Novel mouse model of autosomal semidominant adult hypophosphatasia has a splice site mutation in the tissue nonspecific alkaline phosphatase gene Akp2.
abstract: Deactivating mutations in the TNSALP gene cause HPP. Akp2(-/-) mice model severe infantile HPP, but there is no model for the relatively mild adult form. Here we report on mice with an induced mutation in Akp2 that affects splicing. The phenotype of homozygotes mirror aspects of the adult form of HPP.
  
  
  BACKGROUND
  Hypophosphatasia (HPP) is a clinically varied skeletal disorder resulting from deficiency of tissue nonspecific alkaline phosphatase (TNSALP). Mice lacking Akp2 model infantile HPP characterized by skeletal hypomineralization, impaired growth, seizures, and perinatal mortality. No animal model exists to study the less severe forms of the disease that typically present in later life.
  
  
  METHODS
  N-ethyl-N-nitrosourea (ENU) mutagenesis was used to generate mouse models of human disease. A mouse with low plasma alkaline phosphatase (ALP) activity was identified by our clinical chemistry screen. Its offspring were used for inheritance studies and subjected to biochemical, histological, and radiological phenotyping. DNA was extracted for mapping and osteoblasts harvested for functional studies.
  
  
  RESULTS
  We showed semidominant inheritance of the low ALP phenotype and mapped the underlying point mutation to Akp2. Affected offspring bear the splice site mutation 862 + 5G>A-a hypomorphic allele named Akp2(Hpp). The same mutation has been reported in a patient. Akp2(Hpp/+) mice have approximately 50% of normal plasma ALP but display no other biochemical or skeletal abnormalities. Unlike Akp2(-/-) mice, Akp2(Hpp/Hpp) mice have normal initial skeletal development and growth, a normal lifespan and do not have seizures. TNSALP is low but detectable in Akp2(Hpp/Hpp) plasma. Osteoblasts display approximately 10% of normal ALP activity and reduced intracellular inorganic phosphate levels, yet are capable of normal mineralization in vitro. TNSALP substrates are significantly elevated in urine (inorganic pyrophosphate and phosphoethanolamine) and plasma (pyridoxal 5'-phosphate), whereas plasma inorganic pyrophosphate levels are normal. Akp2(Hpp/Hpp) mice develop late-onset skeletal disease, notably defective endochondral ossification and bone mineralization that leads to arthropathies of knees and shoulders.
  
  
  CONCLUSIONS
  Akp2(Hpp/Hpp) mice mirror a number of clinical features of the human adult form of HPP. These mice provide for the first time an animal model of late onset HPP that will be valuable in future mechanistic studies and for the evaluation of therapies such as those aimed at HPP.
authors.affiliation: Mammalian Genetics Unit, Mary Lyon Centre, Medical Research Council, Harwell, Oxfordshire, United Kingdom. t.hough@har.mrc.ac.uk
authors.forename: Tertius A
authors.initials: TA
authors.lastname: Hough
authors.forename: Monika
authors.initials: M
authors.lastname: Polewski
authors.forename: Kristen
authors.initials: K
authors.lastname: Johnson
authors.forename: Michael
authors.initials: M
authors.lastname: Cheeseman
authors.forename: Patrick M
authors.initials: PM
authors.lastname: Nolan
authors.forename: Lucie
authors.initials: L
authors.lastname: Vizor
authors.forename: Sohaila
authors.initials: S
authors.lastname: Rastan
authors.forename: Alan
authors.initials: A
authors.lastname: Boyde
authors.forename: Kenneth
authors.initials: K
authors.lastname: Pritzker
authors.forename: A Jackie
authors.initials: AJ
authors.lastname: Hunter
authors.forename: Elizabeth M C
authors.initials: EM
authors.lastname: Fisher
authors.forename: Robert
authors.initials: R
authors.lastname: Terkeltaub
authors.forename: Steve D M
authors.initials: SD
authors.lastname: Brown
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary; D004195:Disease Models, Animal; D005799:Genes, Dominant; D007014:Hypophosphatasia / Q000235:genetics*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D012326:RNA Splicing
chemical_list: D018076:DNA, Complementary; C510878:ALPL protein, mouse; D000469:Alkaline Phosphatase