Lisa Hamerlinck, Eva D’haene, Michael Vaughan, Nore Van Loon, María del Rocío Pérez Baca, Sebastian Leimbacher, Michael Kosicki, Lara Colombo, Lukas Genbrugge, Lies Vantomme, Esperanza Daal, Luiza Ramos, Daniela Avdjieva-Tzavella, Himanshu Goel, Koen Devriendt, Albena Jordanova, Annelies Dheedene, Axel Visel, Björn Menten, Bert Callewaert, Sarah Vergult
The FOXG1 transcription factor is a crucial regulator of embryonic brain development. Pathogenic FOXG1 variants cause FOXG1 syndrome. Although structural variants in the non-coding region downstream of FOXG1 have been reported in 38 individuals with similar characteristics, the regulatory pathomechanisms remain unknown. Here, we identify two non-coding structural variants in individuals with FOXG1 syndrome-like features, allowing us to delineate a ~ 124 kb commonly affected regulatory region. Using epigenomic profiling and in vivo enhancer assays, we characterize and validate regulatory elements within the commonly affected regulatory region and wider FOXG1 TAD. We see strong activation of previously validated forebrain enhancers, and identify an enhancer cluster and progenitor-specific enhancer region that are strongly activated during forebrain-directed neural progenitor cell differentiation, a process in which FOXG1 is an important regulator. Perturbation of these elements results in varying degrees of reduced FOXG1 transcription in forebrain neural progenitor cells and in population shifts within these cells, while removal of the TAD boundary leads to aberrant expression of the neighbouring PRKD1 gene. Our findings characterize enhancer and architectural elements essential for proper FOXG1 transcription during neurodevelopment, therefore improving variant interpretation in this region.