Hypophosphatasia (HPP) is the inborn-error-of-metabolism that features rickets or osteomalacia due to loss-of-function mutation within the tissue-nonspecific alkaline phosphatase (TNAP) gene. To-date, there is no established medical treatment for this condition. Our work has clearly shown that TNAP knockout mice (Akp2-/- mice) faithfully mimic the severe infantile form of HPP, and that their rickets/osteomalacia is caused by accumulation of extracellular levels of inorganic pyrophosphate (ePPi), one of TNAP's natural substrates and a potent calcification inhibitor. In turn, high ePPi leads to a secondary elevation of skeletal osteopontin (OPN), which functions as another potent calcification inhibitor. In turn, overexpression of TNAP in skeletal tissues leads to increases in bone mineral density and bone volume fraction, via a mechanism that involves reduction in ePPi levels and increased dephosphorylation of skeletal OPN. Epileptic seizures lead to the early demise of Akp2-/- mice and are caused by inadequate utilization of pyridoxal-5'-phosphate (PLP - a hydrophilic form of Vitamin B6), another natural substrate of TNAP. In a major breakthrough during this last funding period, we have shown that enzyme replacement therapy (EzRT) using a bone-targeted form of TNAP is able to completely prevent all the symptoms of HPP in the Akp2-/- mice, including the epileptic seizures and the severe skeletal and dental abnormalities characteristic of this excellent model of infantile HPP. Our findings represent the first successful use of EzRT for a heritable primary disease of the skeleton, and are a foundation for future therapeutic trials for human HPP. With that ultimate goal in mind, the present competitive renewal application focuses sharply on optimizing this EzRT strategy in TNAP-deficient mouse models.
Our Specific Aims are: I) To test the hypothesis that EzRT not only prevents HPP in Akp2-/- mice but can also rescue Akp2-/- mice with advanced HPP disease. Additionally, we will compare the relative efficacy of administering bone-targeted versus soluble TNAP in Akp2-/- mice. II) To test the hypothesis that EzRT will also be efficacious to prevent and treat """"""""adult HPP"""""""". We will compare the phenotypic abnormalities of three different mouse strains, i.e., a bone-specific TNAP knockout model (Akp2flox/flox;Col1a1-Cre), a transgenic model of dominant HPP (Col1a1- TnapD361V) and a ENU-mutagenesis model of semi-dominant HPP (Akp2Hpp/Hpp) and evaluate the efficacy of EzRT on the most representative model. III) To test the hypothesis that pharmacological """"""""activators"""""""" of TNAP's pyrophosphatase activity can be used to stimulate bone mineralization in vitro and in vivo. We will also assess the ability of these TNAP activators to stimulate in vitro the residual TNAP activity present in serum samples from genotyped HPP patients to identify what TNAP mutations might respond to such a pharmacological intervention. Our work will have an immediate impact on the clinical management of HPP patients, while providing further insights into the pathogenesis of this variable disease. The novel TNAP activators that we have discovered may prove of interest for the prevention and treatment of osteoporosis.

Public Health Relevance

Our studies have shown that sustained delivery of bone-targeted alkaline phosphatase can prevent the sequelae of the heritable form of rickets and osteomalacia known as hypophosphatasia, recapitulated in Akp2-/- mice. These observations represent the first demonstration of successful enzyme replacement therapy to prevent a primary skeletal disease of genetic origin in a mouse model, and are a foundation for clinical trials in patients suffering from this inborn-error-of-metabolism. The work in this application will optimize the use of enzyme replacement therapy to treat the most severe as well as the milder forms of the disease in diverse mouse models. These studies will be followed immediately by clinical trials in human hypophosphatasia patients. In addition, we have uncovered novel activators of alkaline phosphatase function that can be developed into pharmaceuticals to increase bone mineral density in patients with hypophosphatasia and osteoporosis.

National Institute of Health (NIH)
National Institute of Dental & Craniofacial Research (NIDCR)
Research Project (R01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-MOSS-L (04))
Program Officer
Wan, Jason
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Sanford-Burnham Medical Research Institute
La Jolla
United States
Zip Code
Pettengill, Matthew; Matute, Juan D; Tresenriter, Megan et al. (2018) Correction: Human alkaline phosphatase dephosphorylates microbial products and is elevated in preterm neonates with a history of late-onset sepsis. PLoS One 13:e0197532
Li, Qiaoli; Huang, Jianhe; Pinkerton, Anthony B et al. (2018) Inhibition of Tissue-Nonspecific Alkaline Phosphatase Attenuates Ectopic Mineralization in the Abcc6-/- Mouse Model of PXE but Not in the Enpp1 Mutant Mouse Models of GACI. J Invest Dermatol :
Yang, Won Ho; Heithoff, Douglas M; Aziz, Peter V et al. (2018) Accelerated Aging and Clearance of Host Anti-inflammatory Enzymes by Discrete Pathogens Fuels Sepsis. Cell Host Microbe 24:500-513.e5
Patel, Jessal J; Zhu, Dongxing; Opdebeeck, Britt et al. (2018) Inhibition of arterial medial calcification and bone mineralization by extracellular nucleotides: The same functional effect mediated by different cellular mechanisms. J Cell Physiol 233:3230-3243
Simão, Ana Maria Sper; Bolean, Maytê; Favarin, Bruno Zoccaratto et al. (2018) Lipid microenvironment affects the ability of proteoliposomes harboring TNAP to induce mineralization without nucleators. J Bone Miner Metab :
Foster, B L; Ao, M; Salmon, C R et al. (2018) Osteopontin regulates dentin and alveolar bone development and mineralization. Bone 107:196-207
Brun, Lucas R; Lombarte, M; Roma, S et al. (2018) Increased calcium uptake and improved trabecular bone properties in intestinal alkaline phosphatase knockout mice. J Bone Miner Metab 36:661-667
Bottini, Massimo; Mebarek, Saida; Anderson, Karen L et al. (2018) Matrix vesicles from chondrocytes and osteoblasts: Their biogenesis, properties, functions and biomimetic models. Biochim Biophys Acta Gen Subj 1862:532-546
Morcos, M W; Al-Jallad, H; Li, J et al. (2018) PHOSPHO1 is essential for normal bone fracture healing: An Animal Study. Bone Joint Res 7:397-405
Huang, Nai-Jia; Lin, Ying-Cing; Lin, Chung-Yueh et al. (2018) Enhanced phosphocholine metabolism is essential for terminal erythropoiesis. Blood 131:2955-2966

Showing the most recent 10 out of 109 publications