Functional and mechanistic characterization of YWHAZ variants associated with human diseases

最近基因组医学研究的爆炸性增长导致越来越多的 先天缺陷患者的基因组变异。然而，详细的分析表明 影响发育过程的变异很少，导致分类的变异越来越多 作为不确定意义的变体(VU)。了解VU的功能后果将有所帮助 患者、他们的家人和他们的医生了解他们病情的遗传基础，使 临床医生提供个性化的医疗护理和治疗，并扩大我们的生物学知识 以及疾病过程中涉及的分子的机械性操作。在这份提案中，我们计划 采用多学科方法研究与先天性疾病相关的YWHAZ变异 综合症。我们对一例类风湿关节炎患者的一种变异体的初步研究揭示 在脊椎动物中，该变体比野生型YWHAZ更有效地激活RAF-ERK途径 动物模型，非洲爪蛙非洲爪蛙。结果首次表明，YWHAZ 变异体可能与Rasopathies的病因学有关。其他几个YWHAZ变种也与 人类疾病，但变异体的功能意义尚未得到检验。我们的建议 研究将利用非洲爪哇模式的优势，生化和结构的力量 研究，以及小鼠遗传系统的力量，以详细描述YWHAZ 变种。我们将调查YWHAZ变体是否以及如何改变了活动(目标1)，如何 序列变异影响蛋白质结构和相互作用(目标2)，以及变异如何诱导 小鼠模型的病理学(目标3)。我们的调查团队与 不同研究学科的专业知识有望产生对疾病的深刻洞察- 相关的YWHAZ基因的功能和机制。

Recent explosion in genomic medicine studies has led to identification of an increasing number of genomic variations from patients with congenital defects. However, detailed analyses about how the variants affect developmental processes are scarce, resulting in a growing class of variants classified as variants of uncertain significance (VUS). Understanding functional consequences of VUS will help patients, their families, and their doctors to learn genetic underpinnings of their conditions, enable clinicians to provide personalized medical care and treatment, and expand our knowledge on biology and mechanistic operations of the molecules involved in disease processes. In this proposal, we plan to employ multidisciplinary approaches to investigate the YWHAZ variants associated with congenital syndromes. Our preliminary studies on one variant identified from a patient with RASopathy revealed that the variant activated the RAF-ERK pathway more efficiently than wild type YWHAZ in a vertebrate animal model, the African clawed frog Xenopus. The results show for the first time that YWHAZ variant may contribute to etiology of RASopathies. Several other YWHAZ variants also associate with human disorders, but functional significance of the variants has not been examined. Our proposed research will leverage the advantages of the Xenopus model, the power of biochemical and structural studies, and the strength of the mouse genetic system for detailed characterization of the YWHAZ variants. We will investigate whether and how YWHAZ variants have altered activities (aim 1), how sequence variations affect protein structure and interaction (aim 2), and how the variants induce pathology in the mouse models (aim 3). The novel combination of our investigation teams with expertise in distinct research disciplines promises generation of profound insight into disease- associated YWHAZ gene function and mechanisms.