Zebrafish model of Peters-plus syndrome

描述(申请人提供)：彼得斯综合征(PPS)是一种常染色体隐性遗传的变异性复杂疾病。PPS常见的特征有：眼前段缺陷、身材矮小、短指、唇裂和/或腭裂、各种头面部缺陷，以及脑、心脏和泌尿生殖系统异常。PPS综合征最近被发现是由编码糖基转移酶的B3GALTL基因突变引起的。糖基转移酶在各种生物过程中发挥着重要作用，是催化糖分子与特定受体结合的酶。PPS患者的B3GALTL有两个突变的等位基因，通常要么基因完全缺失，要么核苷酸改变会导致截短的蛋白质缺乏催化结构域。B3GALTL突变似乎可以解释大多数经典的PPS，但在高度相似的疾病患者中还没有发现该基因的突变。目前，PPS的发生机制及相关条件尚不清楚，B3GALTL缺乏症/PPS的动物模型尚未建立。研究人员的初步研究已经在斑马鱼中发现了两个b3galtl基因，它们表现出与人类PPS特征一致的表达模式和敲除表型(B3galtlb)。由于PPS具有高度可变的表型特征，因此建立PPS的斑马鱼模型对于该疾病的遗传解剖和其他致病或修饰基因的鉴定是必不可少的。在这项应用中，她计划在特定目标1中重点确定斑马鱼b3galtl基因的功能序列和表达模式，其中包括转录本、基因组结构、功能调节区和b3galtl基因表达模式的特征；在特定目标2中，通过吗啡介导的单基因敲除和双基因敲除分析来检查与b3galtl基因缺陷相关的表型。将利用锌指核酸酶技术建立斑马鱼品系，以获得表现出稳健表型的基因(S)，以创建脊椎动物模型(S)，以研究与B3GalTL/b3galtl缺乏症相关的复杂和可变特征。 简介：彼得斯综合征(PPS)是一种复杂的疾病，涉及眼、头、脑、手、心脏、肾脏和身高的异常发育。PPS的机制(S)和相关的衰弱条件在很大程度上是未知的。B3GALTL基因突变可以解释很大一部分患者的这些表型。对动物模型的研究可以更好地理解潜在的发育机制和其他因素的参与。在这项应用中，将探索一个经典的脊椎动物发育模型-斑马鱼，以深入了解斑马鱼b3galtl基因的相关发育过程，以更好地了解PPS的机制和人类的相关情况。

DESCRIPTION (Provided by Applicant): Peters-plus syndrome (PPS) is a variable complex disorder with autosomal recessive inheritance. The following features are commonly seen in PPS: ocular anterior segment defects, short stature, brachydactyly, cleft lip and/or cleft palate, various craniofacial defects, and brain, heart, as well as genitourinary anomalies. PPS syndrome was recently discovered to be caused by mutations in the B3GALTL gene that encodes a glycosyltransferase. Glycosyltransferases are known to play important roles in various biological processes, and represent enzymes that catalyze the attachment of a sugar molecule to specific acceptor sites. Patients with PPS have two mutated alleles of B3GALTL, typically either a complete deletion of the gene or nucleotide changes predicted to result in a truncated protein lacking its catalytic domain. B3GALTL mutations appear to explain the majority of classic PPS, but no mutations in this gene have been identified in human patients with highly similar disorders. At this time, the developmental mechanisms of PPS and related conditions are not known, and no animal models have been established for B3GALTL-deficiency/PPS. The investigator's preliminary studies have identified two b3galtl genes in the zebrafish that demonstrate expression patterns and knockdown phenotype (for b3galtlb) consistent with PPS features in humans. Since PPS is characterized by a highly variable phenotype, generation of the zebrafish model of PPS is essential to genetic dissection of this disease and identification of additional causative or modifying loci/genes. In this application, she plans to focus, in Specific Aim 1, to determine the functional sequences and expression patterns of zebrafish b3galtl genes, which involves characterization of transcripts, genomic structures, functional regulatory regions, and expression patterns of the b3galtl genes; and in Specific Aim 2, to examine phenotypes associated with deficiency for the b3galtl genes through analyses of morpholino-mediated single as well as double gene knockdowns. Generation of zebrafish lines via zinc-finger nuclease technology will be initiated for gene(s) that demonstrate a robust phenotype to create vertebrate model(s) to study the complex and variable features associated with B3GALTL/b3galtl deficiency. NARRATIVE: Peters-plus syndrome (PPS) is a complex disorder that involves abnormal development of the eye, head, brain, hands, heart, kidney, and stature. Mechanism(s) of PPS and related debilitating conditions are largely unknown. Mutations in B3GALTL gene were shown to explain a significant portion of patients with these phenotypes. Studies in animal models can provide a better understanding of underlying developmental mechanisms and involvement of additional factors. In this application, a classic vertebrate developmental model, zebrafish, will be explored to gain insight into developmental processes associated with orthologous b3galtl genes in zebrafish to better understand mechanisms of PPS and related conditions in humans.