Overexpression of human PHEX under the human beta-actin promoter does not fully rescue the Hyp mouse phenotype.
XLH in humans and the Hyp phenotype in mice are caused by inactivating Phex mutations. Overexpression of human PHEX under the human beta-actin promoter in Hyp mice rescued the bone phenotype almost completely, but did not affect phosphate homeostasis, suggesting that different, possibly independent, pathophysiological mechanisms contribute to hyperphosphaturia and bone abnormalities in XLH.


BACKGROUND
Mutations in PHEX, a phosphate-regulating gene with homologies to endopeptidases on the X chromosome, are responsible for X-linked hypophosphatemia (XLH) in humans, and its mouse homologs, Hyp, Phex(Hyp-2J), Phex(Hyp-Duk), Gy, and Ska1. PHEX is thought to inactivate a phosphaturic factor, which may be fibroblast growth factor 23 (FGF)-23. Consistent with this hypothesis, FGF-23 levels were shown to be elevated in most patients with XLH and in Hyp mice. The aim of this study was, therefore, to examine whether transgenic overexpression of PHEX under the human beta-actin promoter would rescue the Hyp phenotype.


METHODS
We tested this hypothesis by generating two mouse lines expressing human PHEX under the control of a human beta-actin promoter (PHEX-tg). With the exception of brain, RT-PCR analyses showed transgene expression in all tissues examined. PHEX protein, however, was only detected in bone, muscle, lung, skin, and heart. To assess the role of the mutant PHEX, we crossed female heterozygous Hyp mice with male heterozygous PHEX-tg mice to obtain wildtype (WT), PHEX-tg, Hyp, and Hyp/PHEX-tg offspring, which were examined at 3 months of age.


RESULTS
PHEX-tg mice exhibited normal bone and mineral ion homeostasis. Hyp mice showed the known phenotype with reduced body weight, hypophosphatemia, hyperphosphaturia, and rickets. Hyp/PHEX-tg mice had almost normal body weight relative to WT controls, showed a dramatic improvement in femoral BMD, almost normal growth plate width, and, despite remaining disturbances in bone mineralization, almost normal bone architecture and pronounced improvements of osteoidosis and of halo formation compared with Hyp mice. However, Hyp and Hyp/PHEX-tg mice had comparable reductions in tubular reabsorption of phosphate and were hypophosphatemic relative to WT controls.


CONCLUSIONS
Our data suggest that different, possibly independent, pathophysiological mechanisms contribute to renal phosphate wasting and bone abnormalities in Hyp and XLH.
Erben
Reinhold G
RG
Institute of Physiology, Physiological Chemistry and Animal Nutrition, Ludwig Maximilians University, Munich, Germany.
Mayer
Dagmar
D
Weber
Karin
K
Jonsson
Kenneth
K
Jüppner
Harald
H
Lanske
Beate
B
D000199:Actins / Q000235:genetics*; D000818:Animals; D015519:Bone Density / Q000235:genetics; D000089703:Fibroblast Growth Factor-23; D006801:Humans; D017674:Hypophosphatemia / Q000235:genetics* / Q000378:metabolism; D007668:Kidney / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D053402:PHEX Phosphate Regulating Neutral Endopeptidase; D010641:Phenotype; D010710:Phosphates / Q000097:blood / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011506:Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D014018:Tissue Distribution; D015533:Transcriptional Activation; D019076:Transgenes
D000199:Actins; C000717427:FGF23 protein, human; C000717428:Fgf23 protein, mouse; D010710:Phosphates; D011506:Proteins; D000089703:Fibroblast Growth Factor-23; D053402:PHEX Phosphate Regulating Neutral Endopeptidase; C506231:PHEX protein, human; C506232:Phex protein, mouse