Developing a zebrafish model for a novel human neurodevelopmental disorder.

我们最近发现了一种由EIF 5A 1基因突变引起的新型人类疾病，导致儿童发育问题，头部尺寸小和颅面缺陷[1]。EIF 5A 1是独一无二的，因为它是唯一已知的含有氨基酸羟腐胺赖氨酸的蛋白质，并且对于一组重要蛋白质的正确合成至关重要[2]。使用酵母和斑马鱼模型系统，我们已经展示了EIF 5A 1突变如何导致eIF 5A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，我们已经表明特定的食物补充剂可能是这种疾病的潜在治疗方法[1]。本研究将利用原位杂交和荧光显微镜技术研究eIF 5A 1在斑马鱼发育过程中的正常表达模式。CRISPR/Cas将用于产生具有人类EIF 5A 1突变的斑马鱼品系。这些鱼将被表征为脑和颅面缺陷，并将使用质谱法，RNA-seq和eIF 5A功能的生化测定来研究它们的组织。将进行突变斑马鱼的表型拯救研究。将建立一个国际患者登记处，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。结果学生将在基础生物学和转化精确医学的界面上生成并验证一种新型人类疾病的脊椎动物模型。学生将学习几个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.