Subjects
The clinical studies were approved by Oxfordshire Research Ethics Committee B (reference C02.143), London Riverside Research Ethics Committee (reference 09/H0706/20), and the Medical Ethical Committee of the Erasmus University Medical Center Rotterdam (MEC-2012-140 and MEC-2013-547). Written informed consent to obtain samples for genetics research was obtained from each child’s parent or guardian. Venous blood was used for DNA extraction and fibroblast cultures were established from skin biopsies taken from scalp incisions during surgical intervention. Intracranial pressures in subject 1 were documented by 24–48 hr direct recording with an intraparenchymal Codman Microsensor.21 The screening panel comprised samples from 307 individuals with syndromic or non-syndromic craniosynostosis. All DNA samples were previously tested for mutation hotspots in FGFR2, FGFR3, TWIST1, and TCF12.3,4 Significant chromosome aneuploidy in individuals with ZIC1 mutations was excluded by karyotyping and/or array comparative genomic hybridization. Where necessary, correct biological relationships were confirmed by segregation analysis of a panel of 13 microsatellites (D1S2868, D3S1311, D4S403, D5S2027, D6S1610, D7S519, D9S158, D10S548, D11S898, D13S1265, D14S280, D16S415, and D18S474).