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Craniosynostosis-associated Fgfr2(C342Y) mutant bone marrow stromal cells exhibit cell autonomous abnormalities in
1Department of Orthodontics and Pediatric Dentistry, School of Dentistry, University of Michigan, Ann Arbor, MI 48109-1078, USA.
Abstract:
We recently reported that cranial bones of Fgfr2(C342Y/+) craniosynostotic mice are diminished in density when compared to those of wild type mice, and that cranial bone cells isolated from the mutant mice exhibit inhibited late stage osteoblast differentiation. To provide further support for the idea that craniosynostosis-associated Fgfr mutations lead to cell autonomous defects in osteoblast differentiation and mineralized tissue formation, here we tested bone marrow stromal cells isolated from Fgfr2(C342Y/+) mice for their ability to differentiate into osteoblasts. Additionally, to determine if the low bone mass phenotype of Crouzon syndrome includes the appendicular skeleton, long bones were assessed by micro CT. Fgfr2(C342Y/+) cells showed increased osteoblastic gene expression during early osteoblastic differentiation but decreased expression of alkaline phosphatase mRNA and enzyme activity, and decreased mineralization during later stages of differentiation, when cultured under 2D in vitro conditions. Cells isolated from Fgfr2(C342Y/+) mice also formed less bone when allowed to differentiate in a 3D matrix in vivo. Cortical bone parameters were diminished in long bones of Fgfr2(C342Y/+) mice. These results demonstrate that marrow stromal cells of Fgfr2(C342Y/+) mice have an autonomous defect in osteoblast differentiation and bone mineralization, and that the Fgfr2(C342Y) mutation influences both the axial and appendicular skeletons.
Insights
Fibroblast growth factor receptor 2 (Fgfr2) mutations in craniosynostosis mice cause intrinsic defects in bone cell differentiation and mineralization. This impacts both skull and long bone development, affecting the axial and appendicular skeletons.
Area of Science:
- Skeletal Biology
- Developmental Biology
- Genetics
Background:
- Craniosynostosis is associated with Fibroblast growth factor receptor 2 (Fgfr2) mutations.
- Previous studies showed diminished cranial bone density and inhibited osteoblast differentiation in Fgfr2(C342Y/+) mice.
- Cell-autonomous defects in osteoblast differentiation and mineralization are hypothesized.
Purpose of the Study:
- To investigate cell-autonomous defects in osteoblast differentiation and bone mineralization in Fgfr2(C342Y/+) mice.
- To determine if the appendicular skeleton is affected in Crouzon syndrome models.
Main Methods:
- Isolation and in vitro culture of bone marrow stromal cells from Fgfr2(C342Y/+) and wild-type mice.
- Assessment of osteoblast differentiation markers (gene expression, alkaline phosphatase activity, mineralization) under 2D culture.
- In vivo assessment of bone formation using a 3D matrix.
- Micro computed tomography (micro CT) analysis of long bones.
Main Results:
- Fgfr2(C342Y/+) cells exhibited increased early osteoblastic gene expression but decreased alkaline phosphatase activity and mineralization in vitro.
- Cells from mutant mice formed less bone in a 3D matrix in vivo.
- Diminished cortical bone parameters were observed in the long bones of Fgfr2(C342Y/+) mice.
Conclusions:
- Marrow stromal cells from Fgfr2(C342Y/+) mice possess an autonomous defect in osteoblast differentiation and bone mineralization.
- The Fgfr2(C342Y) mutation affects both axial and appendicular skeletal development.
