PDF(Pubmed)
Abstract：
UNASSIGNED: The purpose of this study is to report five novel FZD4 mutations identified in familial exudative vitreoretinopathy (FEVR) and to analyze and summarize the pathogenic mechanisms of 34 of 96 reported missense mutations in FZD4.
UNASSIGNED: Five probands diagnosed with FEVR and their family members were enrolled in the study. Ocular examinations and targeted gene panel sequencing were conducted on all participants. Plasmids, each carrying 29 previously reported FZD4 missense mutations and five novel mutations, were constructed based on the selection of mutations from each domain of FZD4. These plasmids were used to investigate the effects of mutations on protein expression levels, Norrin/β-catenin activation capacity, membrane localization, norrin binding ability, and DVL2 recruitment ability in HEK293T, HEK293STF, and HeLa cells.
UNASSIGNED: All five novel mutations (S91F, V103E, C145S, E160K, C377F) responsible for FEVR were found to compromise Norrin/β-catenin activation of FZD4 protein. After reviewing a total of 34 reported missense mutations, we categorized all mutations based on their functional changes: signal peptide mutations, cysteine mutations affecting disulfide bonds, extracellular domain mutations influencing norrin binding, transmembrane domain (TM) 1 and TM7 mutations impacting membrane localization, and intracellular domain mutations affecting DVL2 recruitment.
UNASSIGNED: We expanded the spectrum of FZD4 mutations relevant to FEVR and experimentally demonstrated that missense mutations in FZD4 can be classified into five categories based on different functional changes.
UNASSIGNED: Five probands diagnosed with FEVR and their family members were enrolled in the study. Ocular examinations and targeted gene panel sequencing were conducted on all participants. Plasmids, each carrying 29 previously reported FZD4 missense mutations and five novel mutations, were constructed based on the selection of mutations from each domain of FZD4. These plasmids were used to investigate the effects of mutations on protein expression levels, Norrin/β-catenin activation capacity, membrane localization, norrin binding ability, and DVL2 recruitment ability in HEK293T, HEK293STF, and HeLa cells.
UNASSIGNED: All five novel mutations (S91F, V103E, C145S, E160K, C377F) responsible for FEVR were found to compromise Norrin/β-catenin activation of FZD4 protein. After reviewing a total of 34 reported missense mutations, we categorized all mutations based on their functional changes: signal peptide mutations, cysteine mutations affecting disulfide bonds, extracellular domain mutations influencing norrin binding, transmembrane domain (TM) 1 and TM7 mutations impacting membrane localization, and intracellular domain mutations affecting DVL2 recruitment.
UNASSIGNED: We expanded the spectrum of FZD4 mutations relevant to FEVR and experimentally demonstrated that missense mutations in FZD4 can be classified into five categories based on different functional changes.
摘要：
■这项研究的目的是报告在家族性渗出性玻璃体视网膜病变（FEVR）中发现的5个新的FZD4突变，并分析和总结FZD4中96个报告的错义突变中的34个的致病机制。
■5名被诊断为FEVR的先证者及其家庭成员被纳入研究。对所有参与者进行眼部检查和靶向基因组测序。质粒,每个携带29个先前报道的FZD4错义突变和5个新突变，基于从FZD4的每个结构域中选择突变来构建。这些质粒用于研究突变对蛋白质表达水平的影响，Norrin/β-catenin激活能力，膜定位，Norrin结合能力，和DVL2在HEK293T中的招募能力，HEK293STF,HeLa细胞
■所有五个新突变（S91F，V103E,C145S,E160K,发现负责FEVR的C377F）损害了FZD4蛋白的Norrin/β-catenin激活。在回顾了总共34个报告的错义突变后，我们根据功能变化对所有突变进行分类：信号肽突变，影响二硫键的半胱氨酸突变，影响norrin结合的细胞外结构域突变，跨膜结构域（TM）1和TM7突变影响膜定位，和细胞内结构域突变影响DVL2募集。
■我们扩展了与FEVR相关的FZD4突变谱，并通过实验证明了FZD4中的错义突变可以根据不同的功能变化分为五类。
■5名被诊断为FEVR的先证者及其家庭成员被纳入研究。对所有参与者进行眼部检查和靶向基因组测序。质粒,每个携带29个先前报道的FZD4错义突变和5个新突变，基于从FZD4的每个结构域中选择突变来构建。这些质粒用于研究突变对蛋白质表达水平的影响，Norrin/β-catenin激活能力，膜定位，Norrin结合能力，和DVL2在HEK293T中的招募能力，HEK293STF,HeLa细胞
■所有五个新突变（S91F，V103E,C145S,E160K,发现负责FEVR的C377F）损害了FZD4蛋白的Norrin/β-catenin激活。在回顾了总共34个报告的错义突变后，我们根据功能变化对所有突变进行分类：信号肽突变，影响二硫键的半胱氨酸突变，影响norrin结合的细胞外结构域突变，跨膜结构域（TM）1和TM7突变影响膜定位，和细胞内结构域突变影响DVL2募集。
■我们扩展了与FEVR相关的FZD4突变谱，并通过实验证明了FZD4中的错义突变可以根据不同的功能变化分为五类。
