Successful gene therapy in utero for lethal murine hypophosphatasia.

Hypophosphatasia (HPP), caused by mutations in the gene ALPL encoding tissue-nonspecific alkaline phosphatase (TNALP), is an inherited systemic skeletal disease characterized by mineralization defects of bones and teeth. The clinical severity of HPP varies widely, from a lethal perinatal form to mild odontohypophosphatasia showing only dental manifestations. HPP model mice (Akp2(-/-)) phenotypically mimic the severe infantile form of human HPP; they appear normal at birth but die by 2 weeks of age because of growth failure, hypomineralization, and epileptic seizures. In the present study, we investigated the feasibility of fetal gene therapy using the lethal HPP model mice. On day 15 of gestation, the fetuses of HPP model mice underwent transuterine intraperitoneal injection of adeno-associated virus serotype 9 (AAV9) expressing bone-targeted TNALP. Treated and delivered mice showed normal weight gain and seizure-free survival for at least 8 weeks. Vector sequence was detected in systemic organs including bone at 14 days of age. ALP activities in plasma and bone were consistently high. Enhanced mineralization was demonstrated on X-ray images of the chest and forepaw. Our data clearly demonstrate that systemic injection of AAV9 in utero is an effective strategy for the treatment of lethal HPP mice. Fetal gene therapy may be an important choice after prenatal diagnosis of life-threatening HPP.

[1]  Didier Y. R. Stainier,et al.  Molecular control of endothelial cell behaviour during blood vessel morphogenesis , 2011, Nature Reviews Molecular Cell Biology.

[2]  J. Millán,et al.  Dose response of bone-targeted enzyme replacement for murine hypophosphatasia. , 2011, Bone.

[3]  N. Miyake,et al.  Global gene transfer into the CNS across the BBB after neonatal systemic delivery of single-stranded AAV vectors , 2011, Brain Research.

[4]  J. Millán,et al.  Rescue of severe infantile hypophosphatasia mice by AAV-mediated sustained expression of soluble alkaline phosphatase. , 2011, Human gene therapy.

[5]  P. Zoltick,et al.  Early gestational gene transfer of IL-10 by systemic administration of lentiviral vector can prevent arthritis in a murine model , 2011, Gene Therapy.

[6]  S. Ikegawa,et al.  Prevalence of c.1559delT in ALPL, a common mutation resulting in the perinatal (lethal) form of hypophosphatasia in Japanese and effects of the mutation on heterozygous carriers , 2011, Journal of Human Genetics.

[7]  László Négyessy,et al.  Layer-specific activity of tissue non-specific alkaline phosphatase in the human neocortex , 2011, Neuroscience.

[8]  Jeffrey J. Gray,et al.  Enzyme Replacement Therapy Prevents Dental Defects in a Model of Hypophosphatasia , 2011, Journal of dental research.

[9]  J. Millán,et al.  Prolonged Survival and Phenotypic Correction of Akp2−/− Hypophosphatasia Mice by Lentiviral Gene Therapy , 2010, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[10]  M. Whyte.  Physiological role of alkaline phosphatase explored in hypophosphatasia , 2010, Annals of the New York Academy of Sciences.

[11]  K. Ozono,et al.  Clinical Characteristics of Perinatal Lethal Hypophosphatasia: A Report of 6 Cases , 2010, Clinical pediatric endocrinology : case reports and clinical investigations : official journal of the Japanese Society for Pediatric Endocrinology.

[12]  N. Miyake,et al.  Global diffuse distribution in the brain and efficient gene delivery to the dorsal root ganglia by intrathecal injection of adeno‐associated viral vector serotype 1 , 2009, The journal of gene medicine.

[13]  K. Foust,et al.  Intravascular AAV9 preferentially targets neonatal neurons and adult astrocytes , 2009, Nature Biotechnology.

[14]  R. Terkeltaub,et al.  Enzyme Replacement Therapy for Murine Hypophosphatasia , 2007, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[15]  Daniel G. Miller,et al.  AAV Vector Integration Sites in Mouse Hepatocellular Carcinoma , 2007, Science.

[16]  F. Ozturk,et al.  Male germ-line cells are at risk following direct-injection retroviral-mediated gene transfer in utero. , 2005, Molecular therapy : the journal of the American Society of Gene Therapy.

[17]  Christof von Kalle,et al.  Oncogenesis following delivery of a nonprimate lentiviral gene therapy vector to fetal and neonatal mice. , 2005, Molecular therapy : the journal of the American Society of Gene Therapy.

[18]  D. Kohn,et al.  Fetal gene transfer using lentiviral vectors and the potential for germ cell transduction in rhesus monkeys (Macaca mulatta). , 2005, Human gene therapy.

[19]  B. Långström,et al.  Pyridoxine effect on synthesis rate of serotonin in the monkey brain measured with positron emission tomography , 2005, Journal of Neural Transmission / General Section JNT.

[20]  E. Lehtonen,et al.  Evaluation of risks related to the use of adeno-associated virus-based vectors. , 2003, Current gene therapy.

[21]  T. Shimada,et al.  Bone formation following transplantation of genetically modified primary bone marrow stromal cells , 2003, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[22]  M. Dallman,et al.  In utero gene transfer of human factor IX to fetal mice can induce postnatal tolerance of the exogenous clotting factor. , 2003, Blood.

[23]  G. Lipshutz,et al.  Reexpression following readministration of an adenoviral vector in adult mice after initial in utero adenoviral administration. , 2000, Molecular therapy : the journal of the American Society of Gene Therapy.

[24]  J. Millán,et al.  Alkaline Phosphatase Knock‐Out Mice Recapitulate the Metabolic and Skeletal Defects of Infantile Hypophosphatasia , 1999, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[25]  P. Dijkhuizen,et al.  Purification of recombinant adeno-associated virus by iodixanol gradient ultracentrifugation allows rapid and reproducible preparation of vector stocks for gene transfer in the nervous system. , 1999, Human gene therapy.

[26]  J. Bergelson,et al.  Temporally regulated expression patterns following in utero adenovirus-mediated gene transfer , 1999, Gene Therapy.

[27]  A. Salvetti,et al.  Factors influencing recombinant adeno-associated virus production. , 1998, Human gene therapy.

[28]  J. Millán,et al.  Inactivation of two mouse alkaline phosphatase genes and establishment of a model of infantile hypophosphatasia , 1997, Developmental dynamics : an official publication of the American Association of Anatomists.

[29]  J. F. Sharp,et al.  Reversal of cystic fibrosis phenotype in mice by gene therapy in utero , 1997, The Lancet.

[30]  S. Sasaki,et al.  Defection of minor immunological differences among human “universal‐type” alkaline phosphatases , 1988, Journal of cellular biochemistry.