Developing a zebrafish model for a novel human neurodevelopmental disorder.

我们最近发现了一种新的人类疾病，由EIF5A1基因突变引起，导致儿童发育问题、头小和颅面缺陷。EIF5A1是独一无二的，因为它是唯一已知的含有氨基酸hypusine的蛋白质，并且对于一组重要蛋白质[2]的正确合成至关重要。利用酵母和斑马鱼模型系统，我们已经展示了EIF5A1突变如何导致eIF5A功能受损并导致与疾病一致的表型。有趣的是，在相同的模型系统中，我们已经证明了一种特定的食物补充剂可能是治疗这种疾病的潜在方法。项目利用原位杂交和荧光显微镜技术研究eIF5A1在发育中的斑马鱼的正常表达模式。CRISPR/Cas将用于产生具有人类EIF5A1突变的斑马鱼品系。这些鱼的大脑和颅面缺陷将被表征，它们的组织将使用质谱、RNA-seq和eIF5A功能的生化分析进行研究。将对突变斑马鱼进行表型拯救研究。将建立一个国际患者注册表，以进一步研究人类表型，并为未来的治疗试验准备一个队列。该学生将在基础生物学和转化精准医学的界面上生成和表征一种新型人类疾病的脊椎动物模型。学生将学习一些基本的跨学科和定量技能，并获得在人类疾病背景下研究整个生物体生理学的重要经验。

We have recently discovered a novel human disorder caused by mutations in the EIF5A1 gene resulting in developmental problems, small head size and craniofacial defects in children [1]. EIF5A1 is unique because it is the only known protein to contain the amino acid hypusine and is essential for proper synthesis of a set of important proteins[2]. Using yeast and zebrafish model systems we have shown how EIF5A1 mutations lead to impaired eIF5A function and cause phenotypes consistent with the disease. Interestingly, in the same model systems we have shown that a specific food supplement may be a potential treatment for this disease[1]. ProjectThe normal expression pattern of eIF5A1 in developing zebrafish will be studied using in-situ hybridisation and fluorescent microscopy. CRISPR/Cas will be used to generate zebrafish lines with human EIF5A1 mutations. These fish will be characterized for brain and craniofacial defects and their tissues will be studied using Mass-spectrometry, RNA-seq and biochemical assays of eIF5A functions. Phenotype rescue studies of the mutant zebrafish will be performed. An international patient registry will be developed for further studying the human phenotype and to prepare a cohort for future treatment trials.Outcome The student will generate and characterise vertebrate models of a novel human disorder at the interface of fundamental biology and translational precision medicine. The student will learn several fundamental interdisciplinary and quantitative skills, as well as gaining significant experience of studying whole organism physiology in context of human disease.