Genetic Evidence of Serum Phosphate-Independent Functions of FGF-23 on Bone

Abstract

Maintenance of physiologic phosphate balance is of crucial biological importance, as it is fundamental to cellular function, energy metabolism, and skeletal mineralization. Fibroblast growth factor-23 (FGF-23) is a master regulator of phosphate homeostasis, but the molecular mechanism of such regulation is not yet completely understood. Targeted disruption of the Fgf-23 gene in mice ( Fgf-23 ^−/− ) elicits hyperphosphatemia, and an increase in renal sodium/phosphate co-transporter 2a (NaPi2a) protein abundance. To elucidate the pathophysiological role of augmented renal proximal tubular expression of NaPi2a in Fgf-23 ^−/− mice and to examine serum phosphate–independent functions of Fgf23 in bone, we generated a new mouse line deficient in both Fgf-23 and NaPi2a genes, and determined the effect of genomic ablation of NaPi2a from Fgf-23 ^−/− mice on phosphate homeostasis and skeletal mineralization. Fgf-23 ^−/−/NaPi2a ^−/− double mutant mice are viable and exhibit normal physical activities when compared to Fgf-23 ^−/− animals. Biochemical analyses show that ablation of NaPi2a from Fgf-23 ^−/− mice reversed hyperphosphatemia to hypophosphatemia by 6 weeks of age. Surprisingly, despite the complete reversal of serum phosphate levels in Fgf-23 ^−/−/NaPi2a ^−/− , their skeletal phenotype still resembles the one of Fgf23 ^−/− animals. The results of this study provide the first genetic evidence of an in vivo pathologic role of NaPi2a in regulating abnormal phosphate homeostasis in Fgf-23 ^−/− mice by deletion of both NaPi2a and Fgf-23 genes in the same animal. The persistence of the skeletal anomalies in double mutants suggests that Fgf-23 affects bone mineralization independently of systemic phosphate homeostasis. Finally, our data support (1) that regulation of phosphate homeostasis is a systemic effect of Fgf-23, while (2) skeletal mineralization and chondrocyte differentiation appear to be effects of Fgf-23 that are independent of phosphate homeostasis.

Author Summary

Regulation of phosphate homeostasis is a tightly controlled hormonal process involving the intestine, kidneys, and bone, and imbalance of this homeostasis may influence overall mineralization. Fibroblast growth factor-23 (FGF-23) is a circulating hormone produced in the bone that mainly targets the kidneys to control the activity of the sodium/phosphate co-transporters NaPi2a and NaPi2c. These transporters are responsible for actively reabsorbing phosphate ions into the body to maintain physiological serum phosphate levels. Changes in FGF-23 activity lead to human disorders associated with either phosphate wasting or retention. Genetically altered mice in which Fgf-23 activity is lost exhibit severe hyperphosphatemia accompanied by increased NaPi2a activity, and they develop abnormal bone mineralization. Here we describe a new mouse model in which we eliminated NaPi2a from Fgf-23 null mice and show reversal of hyperphosphatemia to hypophosphatemia, suggesting that NaPi2a is the major regulator of phosphate homeostasis. However, the skeletal mineralization defect observed in mice lacking Fgf-23 function remained unchanged in the absence of NaPi2a in these mice. Thus our data indicate that Fgf-23 has a role in controlling bone mineralization independent of systemic phosphate levels.