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RNA interference and inhibition of MEK-ERK signaling prevent abnormal skeletal phenotypes in a mouse model of craniosynostosis


Premature fusion of one or more of the cranial sutures (craniosynostosis) in humans causes over 100 skeletal diseases, which occur in 1 of 2,500 live births1,2,3. Among them is Apert syndrome, one of the most severe forms of craniosynostosis, primarily caused by missense mutations leading to amino acid changes S252W or P253R in fibroblast growth factor receptor 2 (FGFR2)4,5,6. Here we show that a small hairpin RNA targeting the dominant mutant form of Fgfr2 (Fgfr2S252W) completely prevents Apert-like syndrome in mice. Restoration of normal FGFR2 signaling is manifested by an alteration of the activity of extracellular signal-regulated kinases 1 and 2 (ERK1/2), implicating the gene encoding ERK and the genes downstream of it in disease expressivity. Furthermore, treatment of the mutant mice with U0126, an inhibitor of mitogen-activated protein (MAP) kinase kinase 1 and 2 (MEK1/2) that blocks phosphorylation and activation of ERK1/2, significantly inhibits craniosynostosis. These results illustrate a pathogenic role for ERK activation in craniosynostosis resulting from FGFR2 with the S252W substitution and introduce a new concept of small-molecule inhibitor–mediated prevention and therapy for diseases caused by gain-of-function mutations in the human genome.

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Figure 1: Rescue of Apert syndrome–like craniosynostosis by shRNA.
Figure 2: Evaluation of changes in mice treated with shRNA and dynamics of FGFR2 signaling.
Figure 3: Treatment with U0126 repressed Apert syndrome–like craniosynostosis in mice.
Figure 4: ERK phosphorylation and gene expression of wild-type and R2-S252W mutant mice upon treatment with U0126.
Figure 5: Repression of the craniosynostosis phenotype of R2-S252W mutant mice with U0126.


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We thank X. Xu, C. Li and L. Cao for critical discussions. We thank T. Fishler, J. De Soto, J. Miller and A. McPherron for critically reading the manuscript. This work was supported by the Intramural Research Program of the National Institute of Diabetes, Digestive and Kidney Diseases (US National Institutes of Health).

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V.S. performed most experiments, X.C. generated the U6-Fgfr2S252W shRNA transgenic mice and performed shRNA-related experiments, R.-H.W. and H.-S.K. participated in RT-PCR and data analysis and C.-X.D. designed the strategy and wrote the paper.

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Correspondence to Chu-Xia Deng.

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The authors declare no competing financial interests.

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Shukla, V., Coumoul, X., Wang, RH. et al. RNA interference and inhibition of MEK-ERK signaling prevent abnormal skeletal phenotypes in a mouse model of craniosynostosis. Nat Genet 39, 1145–1150 (2007).

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