Dysregulation of Grainyhead‐like 3 expression causes widespread developmental defects

Abstract

The gene encoding the transcription factor, Grainyhead-like 3 (Grhl3), plays critical roles in mammalian development and homeostasis, and these have been uncovered through analysis of loss-of-function models. Grhl3-null embryos exhibit a range of gross phenotypes including a shortened longitudinal axis, thoraco-lumbo-sacral spina bi da and soft-tissue syndactyly. Additional studies reveal that these embryos also exhibit a proliferation/differentiation imbalance in the epidermis. This manifests as a failure in skin barrier formation resulting in peri-natal lethality, and defective wound repair. Conditional inactivation of Grhl3 in the squamous epithelium of adult skin, head and neck tissues, and oesophagus reproduces this proliferation/differentiation imbalance and leads to squamous cell carcinomas. These observations establish GRHL3 as a critical tumour suppressor. Despite these extensive analyses of Grhl3 loss-of-function models, the consequences of gain-of-function of this gene have been di cult to achieve.We have redressed this issue through the generation of a novel mouse model that expresses Grhl3 from a transgene integrated in the Rosa26 locus on an endogenous Grhl3-null background. Expression of the transgene rescues both the neurulation and skin barrier defects of the knockout mice, allowing survival into adulthood. Despite this, the mice are not normal, exhibiting a range of phenotypes attributable to dysregulated Grhl3 expression. In mice homozygous for the transgene, we observe a severe Shaker-Waltzer phenotype associated with hearing impairment. Micro-CT scanning of the cochleae and the vestibular apparatus revealed profound structural alterations underlying these phenotypes. In addition, these mice exhibit other developmental anomalies including hair loss, digit defects and epidermal dysmorphogenesis. These ndings indicate that diverse developmental processes display low tolerance to dysregulation of Grhl3.