Targeted Inactivation of the 25-Hydroxyvitamin D3-1α-Hydroxylase Gene (CYP27B1) Creates an Animal Model of Pseudovitamin D-Deficiency Rickets.

Pseudovitamin D-deficiency rickets is caused by mutations in the cytochrome P450 enzyme, 25-hydroxyvitamin D(3)-1alpha-hydroxylase (1alpha-OHase). Patients with the disease exhibit growth retardation, rickets, and osteomalacia. Serum biochemistry is characterized by hypocalcemia, secondary hyperparathyroidism, and undetectable levels of 1alpha,25-dihydroxyvitamin D(3). We have inactivated the 1alpha-OHase gene in mice after homologous recombination in embryonic stem cells. Serum analysis of homozygous mutant animals confirmed that they were hypocalcemic, hypophosphatemic, hyperparathyroidic, and that they had undetectable 1alpha,25-dihydroxyvitamin D(3). Histological analysis of the bones from 3-week-old mutant animals confirmed the evidence of rickets. At the age of 8 weeks, femurs from 1alpha-OHase-ablated mice present a severe disorganization in the architecture of the growth plate and marked osteomalacia. These results show that we have successfully inactivated the 1alpha-OHase gene in mice and established a valid animal model of pseudovitamin D-deficiency rickets.

[1]  Jonathan T. Wang,et al.  Genetics of Vitamin D 1α-Hydroxylase Deficiency in 17 Families , 1998 .

[2]  H. Koeffler,et al.  Evidence for 1,25-dihydroxyvitamin D3 production by cultured porcine alveolar macrophages , 1991, Molecular and Cellular Endocrinology.

[3]  G. Dickson.  Methods of Calcified Tissue Preparation , 1984 .

[4]  G. Stein,et al.  Osteocalcin gene promoter: Unlocking the secrets for regulation of osteoblast growth and differentiation , 1998, Journal of cellular biochemistry.

[5]  C. Dani,et al.  Post-transcriptional regulation of glyceraldehyde-3-phosphate-dehydrogenase gene expression in rat tissues. , 1984, Nucleic acids research.

[6]  J. Puzas,et al.  Synthesis of 1,25-dihydroxycholecalciferol and 24,25-dihydroxycholecalciferol by calvarial cells. Characterization of the enzyme systems. , 1987, The Biochemical journal.

[7]  B. Milleron,et al.  1,25(OH)2D2 production by T lymphocytes and alveolar macrophages recovered by lavage from normocalcemic patients with tuberculosis. , 1990, The Journal of clinical investigation.

[8]  M. Tassabehji,et al.  Novel Mutations in the 1α‐Hydroxylase (P450c1) Gene in Three Families with Pseudovitamin D–Deficiency Rickets Resulting in Loss of Functional Enzyme Activity in Blood‐Derived Macrophages , 1999, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[9]  J. Pike,et al.  The vitamin D receptor and the syndrome of hereditary 1,25-dihydroxyvitamin D-resistant rickets. , 1999, Endocrine reviews.

[10]  M. Haussler,et al.  The Nuclear Vitamin D Receptor: Biological and Molecular Regulatory Properties Revealed , 1998, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[11]  M. Noda,et al.  Osteopontin expression and function: Role in bone remodeling , 1998, Journal of cellular biochemistry. Supplement.

[12]  J. Puzas,et al.  In vitro synthesis of 1 alpha,25-dihydroxycholecalciferol and 24,25-dihydroxycholecalciferol by isolated calvarial cells. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[13]  M. Seldin,et al.  25‐Hydroxyvitamin D 1α‐Hydroxylase: Structure of the Mouse Gene, Chromosomal Assignment, and Developmental Expression , 2001, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[14]  Tatsuya Yoshizawa,et al.  Mice lacking the vitamin D receptor exhibit impaired bone formation, uterine hypoplasia and growth retardation after weaning , 1997, Nature Genetics.

[15]  J. Roder,et al.  Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[16]  F. Glorieux,et al.  Vitamin D dependency: replacement therapy with calcitriol? , 1981, The Journal of pediatrics.

[17]  A. Craig,et al.  Osteopontin, a transformation-associated cell adhesion phosphoprotein, is induced by 12-O-tetradecanoylphorbol 13-acetate in mouse epidermis. , 1989, The Journal of biological chemistry.

[18]  L. Finberg,et al.  Vitamin D-Dependent Rickets: Actions of Parathyroid Hormone and 25-Hydroxycholecalciferol , 1972, Pediatric Research.

[19]  B. Hogan,et al.  Manipulating the mouse embryo: A laboratory manual , 1986 .

[20]  H. Tenenhouse,et al.  Normal 24-hydroxylation of vitamin D metabolites in patients with vitamin D-dependency rickets type I. Structural implications for the vitamin D hydroxylases. , 1992, The Journal of clinical endocrinology and metabolism.

[21]  M. Noshiro,et al.  Parathyroid hormone inhibits 25-hydroxyvitamin D3-24-hydroxylase mRNA expression stimulated by 1 alpha,25-dihydroxyvitamin D3 in rat kidney but not in intestine. , 1992, The Journal of biological chemistry.

[22]  H. DeLuca,et al.  Serum 1,25-dihydroxyvitamin D levels in normal subjects and in patients with hereditary rickets or bone disease. , 1978, The New England journal of medicine.

[23]  J. Weissenbach,et al.  Linkage disequilibrium analysis in young populations: pseudo-vitamin D-deficiency rickets and the founder effect in French Canadians. , 1996, American journal of human genetics.

[24]  F. Glorieux,et al.  The 25‐Hydroxyvitamin D 1‐Alpha‐Hydroxylase Gene Maps to the Pseudovitamin D‐Deficiency Rickets (PDDR) Disease Locus , 1997, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[25]  H. DeLuca,et al.  Response to Crystalline 1α-Hydroxyvitamin D3 in Vitamin D Dependency , 1975, Pediatric Research.

[26]  S. Kato,et al.  25-Hydroxyvitamin D3 1alpha-hydroxylase and vitamin D synthesis. , 1997, Science.

[27]  H. DeLuca,et al.  Cloning and expression of rat 25-hydroxyvitamin D3-1α-hydroxylase cDNA , 1997 .

[28]  V. Rosen,et al.  Isolation of the human gene for bone gla protein utilizing mouse and rat cDNA clones. , 1986, The EMBO journal.

[29]  W. Miller,et al.  Cloning of human 25-hydroxyvitamin D-1 alpha-hydroxylase and mutations causing vitamin D-dependent rickets type 1. , 1997, Molecular endocrinology.

[30]  S. Kato,et al.  Inactivating mutations in the 25-hydroxyvitamin D3 1alpha-hydroxylase gene in patients with pseudovitamin D-deficiency rickets. , 1998, The New England journal of medicine.

[31]  S. Kato,et al.  No enzyme activity of 25-hydroxyvitamin D3 1alpha-hydroxylase gene product in pseudovitamin D deficiency rickets, including that with mild clinical manifestation. , 1999, The Journal of clinical endocrinology and metabolism.

[32]  W. Pearson,et al.  MOLECULAR CLONING AND SEQUENCING OF CALBINDIN-D9K CDNA FROM MOUSE PLACENTA. , 1988 .

[33]  R. Baron,et al.  Targeted ablation of the vitamin D receptor: an animal model of vitamin D-dependent rickets type II with alopecia. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[34]  F. Glorieux,et al.  Mapping autosomal recessive vitamin D dependency type I to chromosome 12q14 by linkage analysis. , 1990, American journal of human genetics.

[35]  M. Noshiro,et al.  Cloning and expression of cDNA encoding 25‐hydroxyvitamin D3 24‐hydroxylase , 1991, FEBS letters.

[36]  W. Miller,et al.  Vitamin D 1α-Hydroxylase , 2000, Trends in Endocrinology & Metabolism.

[37]  B. Boyan,et al.  Production of 1,25-dihydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 by growth zone and resting zone chondrocytes is dependent on cell maturation and is regulated by hormones and growth factors. , 1992, Endocrinology.

[38]  H. DeLuca,et al.  Current understanding of the molecular actions of vitamin D. , 1998, Physiological reviews.

[39]  W. Miller,et al.  Complete structure of the human gene for the vitamin D 1alpha-hydroxylase, P450c1alpha. , 1997, DNA and cell biology.

[40]  T. Saruta,et al.  Two novel 1α-hydroxylase mutations in French-Canadians with vitamin D dependency rickets type I , 1998 .

[41]  H. DeLuca,et al.  Vitamin D. Basic research and its clinical application. , 1979 .