Functional and mechanistic characterization of YWHAZ variants associated with human diseases

基因组医学研究的最近爆炸导致鉴定越来越多的 先天性缺陷患者的基因组变异。然而，详细分析如何 影响发育过程的变异很少，导致越来越多的变异被分类为 不确定意义的变体（VUS）。了解VUS的功能后果将有助于 患者，他们的家人和他们的医生了解他们病情的遗传基础， 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 和疾病过程中分子的机械操作。在这份提案中，我们计划 采用多学科方法研究与先天性甲状腺炎相关的YWHAZ变异， 综合征我们对一名RAS病患者的一种变异进行了初步研究， 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，非洲爪蟾。结果表明，YWHAZ首次 变异可能有助于RASopathies的病因。其他几个YWHAZ变体也与 人类疾病，但尚未检查变体的功能意义。我们提出的 研究将利用非洲爪蟾模型的优势，生物化学和结构的力量， 研究，以及小鼠遗传系统的强度，以详细描述YWHAZ 变体。我们将研究YWHAZ变体是否以及如何改变活动（目的1）， 序列变异影响蛋白质结构和相互作用（目的2），以及变异如何诱导 小鼠模型中的病理学（目的3）。我们的调查小组与 不同研究学科的专业知识有望产生对疾病的深刻见解- 相关的YWHAZ基因功能和机制。

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.