BRD4 binds to active cranial neural crest enhancers to regulate RUNX2 activity during osteoblast differentiation

Abstract

Cornelia de Lange syndrome (CdLS) is a congenital disorder featuring facial dysmorphism, postnatal growth deficits, cognitive disability, and upper limb abnormalities. CdLS is genetically heterogeneous with cases arising from mutation of BRD4, a bromodomain protein that binds and reads acetylated histones. In this study we have modeled CdLS facial pathology through mouse neural crest cell (NCC) specific mutation of BRD4 to characterize cellular and molecular function in craniofacial development. Mice with BRD4 NCC loss of function died at birth with severe facial hypoplasia, cleft palate, mid-facial clefting, and exencephaly. Following migration, BRD4 mutant NCCs initiated RUNX2 expression for differentiation to osteoblast lineages but failed to induce downstream RUNX2 targets required for lineage commitment. BRD4 bound to active enhancers to regulate expression of osteogenic transcription factors and extracellular matrix components integral for bone formation. RUNX2 physically interacts with a carboxy-terminal (C-terminal) domain in the long isoform of BRD4, can co-occupy osteogenic enhancers, and this association is required for RUNX2 recruitment and appropriate osteoblast differentiation. We conclude that BRD4 controls facial bone development through osteoblast enhancer regulation of the RUNX2 transcriptional program.