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CHD7 cooperates with PBAF to control multipotent neural crest formation

Abstract

Heterozygous mutations in the gene encoding the CHD (chromodomain helicase DNA-binding domain) member CHD7, an ATP-dependent chromatin remodeller homologous to the Drosophila trithorax-group protein Kismet1,2, result in a complex constellation of congenital anomalies called CHARGE syndrome, which is a sporadic, autosomal dominant disorder characterized by malformations of the craniofacial structures, peripheral nervous system, ears, eyes and heart3,4. Although it was postulated 25 years ago that CHARGE syndrome results from the abnormal development of the neural crest, this hypothesis remained untested5. Here we show that, in both humans and Xenopus, CHD7 is essential for the formation of multipotent migratory neural crest (NC), a transient cell population that is ectodermal in origin but undergoes a major transcriptional reprogramming event to acquire a remarkably broad differentiation potential and ability to migrate throughout the body, giving rise to craniofacial bones and cartilages, the peripheral nervous system, pigmentation and cardiac structures6,7. We demonstrate that CHD7 is essential for activation of the NC transcriptional circuitry, including Sox9, Twist and Slug. In Xenopus embryos, knockdown of Chd7 or overexpression of its catalytically inactive form recapitulates all major features of CHARGE syndrome. In human NC cells CHD7 associates with PBAF (polybromo- and BRG1-associated factor-containing complex)8 and both remodellers occupy a NC-specific distal SOX9 enhancer9 and a conserved genomic element located upstream of the TWIST1 gene. Consistently, during embryogenesis CHD7 and PBAF cooperate to promote NC gene expression and cell migration. Our work identifies an evolutionarily conserved role for CHD7 in orchestrating NC gene expression programs, provides insights into the synergistic control of distal elements by chromatin remodellers, illuminates the patho-embryology of CHARGE syndrome, and suggests a broader function for CHD7 in the regulation of cell motility.

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Figure 1: CHD7 knockdown disrupts formation of the migratory, multipotent hNCLCs.
Figure 2: Chd7 is required for expression of transcription factors critical for neural crest specification and migration.
Figure 3: CHD7 interacts with the PBAF complex.
Figure 4: CHD7 and PBAF bind distal regulatory elements upstream of NC transcription factors and synergistically regulate NC gene expression and migration in vivo.

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Acknowledgements

We thank laboratory members for input during the course of this work; S. Brugmann and A. C. Foley for sharing expertise in Xenopus and chick embryology, B. Bedogni, L. Ho and I. Shestopalov for reagents; A. Sanchez-Alvarado for the cartilage staining protocol; G. Crump and S. Cox for sharing unpublished information; and S. Brugmann, E. Duncan, Z. Ma, J. Peng, A. M. Ring and R. A. Roth for comments on the manuscript. This work was supported by CIRM SEED grant RS1-00323, a W. M. Keck Foundation Distinguished Young Scholar Award, a Searle Scholar Award to J.W., an EMBO long-term fellowship to A.R-I., National Institutes of Health (NIH) grant R01DK082664 to Y.Z., March of Dimes grant 6-FY06-335 to J.H., NIH grant R01HL085345 to C-P.C., and an Oak Foundation Fellowship to Y.X.

Author Contributions R.B. developed the in vitro model of human NC formation and designed, performed and interpreted most experiments. D.A.C. did a preliminary characterization of NC migration defects in Xenopus, performed the in situ hybridization analyses of wild-type and CHD7 morphant embryos, and analysed the craniofacial defects in hCHD7 ATPaseK998R tadpoles. A.R-I. performed the genomic analyses of CHD7-Brg1 co-occupancy and contributed ideas. J.Z. and Y.Z. performed the mass spectrometric analysis of CHD7 immunoprecipitates. Y.X. and C-P.C. characterized the heart defects in hCHD7 ATPaseK998R tadpoles. J.H. provided expertise and guidance on in ovo transplantation experiments. T.S. advised on the design and interpretation of Xenopus experiments and contributed reagents. J.W. conceived the project, contributed ideas, interpreted results and wrote the manuscript. All authors edited the manuscript.

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Correspondence to Joanna Wysocka.

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Bajpai, R., Chen, D., Rada-Iglesias, A. et al. CHD7 cooperates with PBAF to control multipotent neural crest formation. Nature 463, 958–962 (2010). https://doi.org/10.1038/nature08733

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