Targeted Disruption of NF1 in Osteocytes Increases FGF23 and Osteoid With Osteomalacia-like Bone Phenotype

Abstract

Neurofibromatosis type 1 (NF1, OMIM 162200), caused by NF1 gene mutations, exhibits multi-system abnormalities, including skeletal deformities in humans. Osteocytes play critical roles in controlling bone modeling and remodeling. However, the role of neurofibromin, the protein product of the NF1 gene, in osteocytes is largely unknown. This study investigated the role of neurofibromin in osteocytes by disrupting Nf1 under the Dmp1-promoter. The conditional knockout (Nf1 cKO) mice displayed serum profile of a metabolic bone disorder with an osteomalacia-like bone phenotype. Serum FGF23 levels were 4 times increased in cKO mice compared with age-matched controls. In addition, calcium-phosphorus metabolism was significantly altered (calcium reduced; phosphorus reduced; parathyroid hormone [PTH] increased; 1,25(OH) D decreased). Bone histomorphometry showed dramatically increased osteoid parameters, including osteoid volume, surface, and thickness. Dynamic bone histomorphometry revealed reduced bone formation rate and mineral apposition rate in the cKO mice. TRAP staining showed a reduced osteoclast number. Micro-CT demonstrated thinner and porous cortical bones in the cKO mice, in which osteocyte dendrites were disorganized as assessed by electron microscopy. Interestingly, the cKO mice exhibited spontaneous fractures in long bones, as found in NF1 patients. Mechanical testing of femora revealed significantly reduced maximum force and stiffness. Immunohistochemistry showed significantly increased FGF23 protein in the cKO bones. Moreover, primary osteocytes from cKO femora showed about eightfold increase in FGF23 mRNA levels compared with control cells. The upregulation of FGF23 was specifically and significantly inhibited by PI3K inhibitor Ly294002, indicating upregulation of FGF23 through PI3K in Nf1-deficient osteocytes. Taken together, these results indicate that Nf1 deficiency in osteocytes dramatically increases FGF23 production and causes a mineralization defect (ie, hyperosteoidosis) via the alteration of calcium-phosphorus metabolism. This study demonstrates critical roles of neurofibromin in osteocytes for osteoid mineralization. © 2017 American Society for Bone and Mineral Research.

The reduced osteogenic potential of Nf1-deficient osteoprogenitors is EGFR-independent

“…This is important to notice because the progressive and long-term nature of tibia bowing and non-union in NF1, and data from genetic mouse models related to this condition, all support the idea that the cell of origin for this condition is a proliferating, undifferentiated mesenchymal progenitor, prior to the expression of Col2 and Osx [10,11]. Hence, the traits and behavior of Nf1 -deficient undifferentiated osteoprogenitors are likely to be more clinically relevant than the characteristics of Nf1 -deficient mature osteoblasts or osteocytes for instance [60], that are unlikely to be ever generated based on the defective differentiation of Nf1 -deficient osteoprogenitors.…”

MEKK2 mediates aberrant ERK activation in neurofibromatosis type I

“…MEKK2 deficiency rescues NF1-associated skeletal phenotypes. Previously, Nf1 fl/fl ;Dmp1-Cre mice have been reported as a model of skeletal NF1, finding that they display spontaneous fractures, accompanied by reduced mechanical strength, low bone mineral density, and high cortical porosity with an osteomalacia-like bone phenotype 25 . Use of the Dmp1-Cre here has the advantage that it avoids the severe growth and joint dysplasia phenotypes associated with the deletion of NF1 in early osteoprogenitors/skeletal stem cells as seen with Prx1 or Col2-Cre, which may complicate interpretation of genetic or pharmacologic rescue experiments 26,27 .…”

Neurofibromatosis type 1 associated with hypophosphatemic osteomalacia due to hypersecretion of fibroblast growth factor 23: a case report

“…A second limitation is that the possibility of increased production of FGF23 from osteocytes cannot be denied. Kamiya et al reported that serum FGF23 levels showed a four-fold increase in NF1 conditional knockout mice (cKO) compared with age-matched controls, and immunohistochemistry showed significantly increased FGF23 protein in the cKO bones [29]. Further evaluations about this should be conducted in future.…”