Optical approaches to delineate novel signalling pathways in skin blistering disease

新一代测序的最新进展为疾病生物学带来了新的见解，也为转化研究提供了机会。通过外显子组测序，我们最近在炎症性皮肤起泡患者中发现了表皮生长因子受体（EGFR）的第一个纯合子功能缺失突变。EGFR信号通路通过与粘附受体整合素家族的串扰，调控细胞的生存、迁移、增殖和分泌等基本过程。然而，控制这种串扰的机制在很大程度上是未知的。该项目将以我们的新发现为基础，测试EGFR和整合素信号之间的平衡在维持皮肤稳态中起关键作用的假设。皮肤是一个复杂的3D组织，如果没有基因操作，分析不同内源性细胞群对组织完整性的贡献是不可能的。“笼化”化合物在天然条件下是惰性的，但在暴露于某些光波长时具有光解活性，这为在特定细胞亚群中操纵内源性蛋白质行为并研究它们在类组织模型中的作用提供了极好的机会。化学笼化是一种潜在的强大的转化研究工具，但迄今为止只应用于非常有限的化合物/目标子集。我们已经与Tocris建立了合作，开发选择性光激活化合物，以研究表皮和真皮等效3D体外模型中表皮和间质室中EGFR和整合素信号之间的串扰。该项目将提供翻译细胞生物学和生物化学方法方面的优秀多学科培训，以研究控制人类皮肤维护的机制。这项研究的数据将为未来受体信号的研究提供重要的新工具，并为导致人类皮肤起泡疾病的基本调控途径提供新的见解。

Recent advances in next generation sequencing have led to new insights into disease biology as well as opportunities for translational research. Through exome sequencing, we recently identified the first homozygous loss of function mutation in epidermal growth factor receptor (EGFR) in patients with inflammatory skin blistering. The EGFR signalling pathway regulates fundamental processes in cells including survival, migration, proliferation and secretion, in part through crosstalk with the integrin family of adhesion receptors. However, the mechanisms that control this crosstalk are largely unknown. This project will build on our novel findings to test the hypothesis that a balance between EGFR and integrin signalling plays a key role in maintaining skin homeostasis. Skin is a complex 3D tissue and analyzing contributions to tissue integrity from different endogenous cell populations is not possible without genetic manipulation. The use of 'caged' compounds, that are inert under native conditions but photolytically activated on exposure to certain light wavelengths, provides an excellent opportunity to manipulate endogenous protein behaviour within specific subsets of cells and study their role in tissue-like models. Chemical caging is a potentially powerful tool for translational research, but has been applied to a very limited subset of compounds/targets to date. We have established a collaboration with Tocris to develop selective optically-activated compounds to investigate the crosstalk between EGFR and integrin signaling within epidermal and stromal compartments of dermal equivalent 3D in vitro models. This project will provide excellent multidisciplinary training in translational cell biology and biological chemistry approaches to study mechanisms that control maintenance of human skin. Data arising from this study will provide important new tools for future studies of receptor signaling, as well as providing novel insight into fundamental regulatory pathways leading to human skin blistering diseases.