Novel Role of a Nucleoporin Gene in Atrial Fibrillation, the Most Common Cardiac

核孔蛋白基因在心房颤动（最常见的心脏疾病）中的新作用

• 批准号: 8442341
• 负责人: QING Kenneth WANG
• 金额: $ 36.99万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-16 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8442341
• 关键词: 5p13; Action Potentials; Affect; Alleles; American; Antibodies; Area; Arrhythmia; Atrial Fibrillation; Binding; Biochemical; Blood Circulation; Cardiac; Catheters; Cell Membrane Permeability; Cell Nucleus; Cell membrane; Cell surface; Cells; Chromosomes; Complex; Computer Simulation; Confocal Microscopy; Cytoplasm; Data; Development; Digitonin; Disease; Docking; Down-Regulation; Drug Targeting; Early Diagnosis; Eukaryotic Cell; Evaluation; Family; Frequencies; Gene Expression; Genes; Genetic; Goals; Heart Atrium; Human; Image Analysis; In Situ; In Vitro; Ion Channel; Knock-out; Knockout Mice; Lead; Link; Macromolecular Complexes; Maps; Measures; Membrane; Messenger RNA; Methods; Mitosis; Modeling; Molecular; Molecular Weight; Morbidity - disease rate; Mus; Muscle Cells; Mutation; Nuclear; Nuclear Envelope; Nuclear Export; Nuclear Import; Nuclear Pore; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Nucleoporin Gene; Organ; Pathogenesis; Pathway interactions; Patients; Pattern; Phenotype; Physiology; Play; Potassium; Property; Proteins; Refractory; Regulation; Regulator Genes; Reporting; Role; Signal Transduction; Sinus; Small Interfering RNA; Structure; Surface; Testing; Therapeutic Intervention; atrioventricular node; base; density; gambogic acid; improved; in vivo; mRNA Export; macromolecule; member; models and simulation; mortality; mutant; novel; protein expression; public health relevance; trafficking

DESCRIPTION (provided by applicant): Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia with more than 2 million Americans affected, and is growing exponentially. The major goal of this project is to identify new molecular determinants and novel molecular mechanisms of AF by molecular characterization of the newly-discovered AF gene NUP155. The NUP155 gene encodes a 155 kDa nucleoporin, which is required for the formation of the nuclear pore complex (NPC) and the assembly of the nuclear envelope during mitosis. The NPC is a large macromolecular complex of about 30 nucleoporins, and plays a key role in bi-directional transport of macromolecules with a molecular weight of >40 kDa across the nucleus membrane. Export of mRNA from the nucleus to the cytoplasm plays an important role in gene expression in eukaryotic cells. The NUP155 protein contains a binding domain that interacts directly with mRNA export factor Gle1, which may anchor GLe1 onto the NPC. NUP155 also interacts directly with a NUP53 which forms further complex with other structural nucleoporins. Thus, NUP155 may play an important role in the assembly of the NPC and regulated control of nuclear export of mRNAs. Mutations in NUP155 cause AF. Two NUP155 mutations have been identified, including mutation R391H reported previously by our group (Zhang et al 2008 Cell) and a newly identified mutation H1104P located within the Gle1 binding domain. Homozygous NUP155-/- knockout (KO) mice die before E8.5, but heterozygous NUP155 mice faithfully recapitulate the human AF phenotype. We have demonstrated that atrial myocytes from NUP155 KO mice show significant shortening of action potential duration (APD). However, the molecular mechanisms by which NUP155 mutations cause APD shortening and consequently AF remain unknown. Based on our new preliminary data that IK1 current densities are increased in NUP155 KO atrial myocytes compared to wild type control myocytes, here we propose that the NPC incorporating a mutant NUP155 subunit or less NUP155, or lacking NUP155 becomes defective structurally and/or functionally. The defective NPC may mis-regulate nuclear export of mRNAs for important atrial ion channel genes and/or their regulatory genes (e.g. genes for IK1 subunits Kir2.1, Kir2.2, Kir2.3 or Kir2.x trafficking factors), which leads to abnormal electrical remodeling of ionic currents in the atria (e.g. IK1). Enhanced IK1 and/or other electrical remodeling cause the shortening of APD and shortening of atrial effective refractory period (ERP), and triggers reentry arrhythmias and AF. To test this hypothesis, we will combine cellular and biochemical approaches, electrophysiological studies, computer modeling, and in vivo KO mouse studies to identify new molecular mechanisms of AF. We will first characterize the AF mutations in NUP155 (R391H, H1104P, NUP155 siRNA mimicking KO allele) for their structural effects on the NPC (interaction with Gle1 and NUP53, and complex formation with other nucleoporins, and nuclear envelope localization) as well as for their functional effects on the NPC (nuclear membrane permeability, nuclear export of mRNAs, nuclear import of proteins using Hsp70 as a marker). Secondly, we will use in vivo intracardiac electrophysiological studies to characterize NUP155 KO mice to assess whether the APD shortening at the cellular level is associated with a shortened atrial ERP and increased inducibility of AF at the organ level. The effects of an IK1 specific blocker, gambogic acid, will be evaluated as potential therapy for AF. Finally, we will evaluate the roles of NUP155 in the nuclear export of mRNAs for IK1 subunits, regulation of cell surface trafficking of IK1 subunits, remodeling of IK1 currents, and effects of IK1 blockers on IK1 currents and atrial APD in NUP155 KO mice. In combination with computer modeling, these studies will investigate the functional impact of down-regulation of NUP155 expression on atrial arrhythmias and identify the substrates and important mechanisms for AF cause by the NUP155 mutations. Results obtained from this study will serve our long-term goal of understanding the cardiac-specific signaling by NUP155 in cardiac physiology and disease.

描述（由申请人提供）：心房颤动 (AF) 是最常见的持续性心律失常，超过 200 万美国人受到影响，并且呈指数级增长。该项目的主要目标是通过新发现的 AF 基因 NUP155 的分子表征来确定 AF 的新分子决定因素和新分子机制。 NUP155 基因编码 155 kDa 核孔蛋白，它是核孔复合体 (NPC) 的形成和有丝分裂过程中核膜组装所必需的。 NPC是由约30个核孔蛋白组成的大分子复合物，在分子量>40 kDa的大分子穿过核膜的双向运输中发挥关键作用。 mRNA从细胞核到细胞质的输出在真核细胞的基因表达中起着重要作用。 NUP155 蛋白含有一个与 mRNA 输出因子 Gle1 直接相互作用的结合域，该因子可能将 GLe1 锚定到 NPC 上。 NUP155 还直接与 NUP53 相互作用，NUP53 与其他结构核孔蛋白形成进一步的复合物。因此，NUP155可能在NPC的组装和mRNA核输出的调节控制中发挥重要作用。 NUP155 突变会导致 AF。已鉴定出两个 NUP155 突变，包括我们小组先前报道的突变 R391H（Zhang 等人 2008 Cell）和新鉴定的位于 Gle1 结合域内的突变 H1104P。纯合 NUP155-/- 敲除 (KO) 小鼠在 E8.5 之前死亡，但杂合 NUP155 小鼠忠实地再现了人类 AF 表型。我们已经证明，NUP155 KO 小鼠的心房肌细胞表现出动作电位持续时间 (APD) 显着缩短。然而，NUP155 突变导致 APD 缩短并进而导致 AF 的分子机制仍不清楚。根据我们新的初步数据，与野生型对照肌细胞相比，NUP155 KO 心房肌细胞中的 IK1 电流密度增加，在此我们提出，含有突变 NUP155 亚基或更少 NUP155 或缺乏 NUP155 的 NPC 在结构和/或功能上变得有缺陷。有缺陷的 NPC 可能会错误调节重要心房离子通道基因和/或其调节基因（例如 IK1 亚基 Kir2.1、Kir2.2、Kir2.3 或 Kir2.x 运输因子的基因）的 mRNA 核输出，从而导致心房中离子电流（例如 IK1）的异常电重塑。 IK1和/或其他电重塑的增强导致APD缩短和心房有效不应期（ERP）缩短，并引发折返性心律失常和AF。 为了验证这一假设，我们将结合细胞和生化方法、电生理学研究、计算机建模和体内 KO 小鼠研究来确定 AF 的新分子机制。我们将首先表征 NUP155 中的 AF 突变（R391H、H1104P、NUP155 siRNA 模拟 KO 等位基因），了解它们对 NPC 的结构影响（与 Gle1 和 NUP53 相互作用、与其他核孔蛋白形成复合物以及核膜定位）以及它们对 NPC 的功能影响（核膜通透性、mRNA 的核输出、 使用 Hsp70 作为标记的蛋白质核输入）。其次，我们将利用体内心内电生理学研究来表征 NUP155 KO 小鼠，以评估细胞水平上的 APD 缩短是否与心房 ERP 缩短和器官水平上 AF 诱导性增加有关。 IK1 特异性阻断剂藤黄酸的效果将作为 AF 的潜在疗法进行评估。最后，我们将评估 NUP155 在 IK1 亚基 mRNA 核输出、IK1 亚基细胞表面运输的调节、IK1 电流重塑以及 IK1 阻滞剂对 NUP155 KO 小鼠中 IK1 电流和心房 APD 的影响中的作用。结合计算机模型，这些研究将探讨 NUP155 表达下调对房性心律失常的功能影响，并确定 NUP155 突变引起 AF 的底物和重要机制。这项研究获得的结果将有助于我们了解 NUP155 在心脏生理学和疾病中的心脏特异性信号传导的长期目标。

项目成果

Functional dominant-negative mutation of sodium channel subunit gene SCN3B associated with atrial fibrillation in a Chinese GeneID population.

• DOI: 10.1016/j.bbrc.2010.06.042
• 发表时间: 2010-07-16
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Wang, Pengyun;Yang, Qinbo;Wu, Xiaofen;Yang, Yanzong;Shi, Lisong;Wang, Chuchu;Wu, Gang;Xia, Yunlong;Yang, Bo;Zhang, Rongfeng;Xu, Chengqi;Cheng, Xiang;Li, Sisi;Zhao, Yuanyuan;Fu, Fenfen;Liao, Yuhua;Fang, Fang;Chen, Qiuyun;Tu, Xin;Wang, Qing K.
• 通讯作者: Wang, Qing K.

Significant association of SNP rs2106261 in the ZFHX3 gene with atrial fibrillation in a Chinese Han GeneID population.

ZFHX3 基因中的 S​​NP rs2106261 与中国汉族 GeneID 人群中心房颤动的显着相关性

• DOI: 10.1007/s00439-010-0912-6
• 发表时间: 2011-03
• 期刊: Human genetics
• 影响因子: 5.3
• 作者: Li C;Wang F;Yang Y;Fu F;Xu C;Shi L;Li S;Xia Y;Wu G;Cheng X;Liu H;Wang C;Wang P;Hao J;Ke Y;Zhao Y;Liu M;Zhang R;Gao L;Yu B;Zeng Q;Liao Y;Yang B;Tu X;Wang QK
• 通讯作者: Wang QK

BRG1 variant rs1122608 on chromosome 19p13.2 confers protection against stroke and regulates expression of pre-mRNA-splicing factor SFRS3.

• DOI: 10.1007/s00439-013-1389-x
• 发表时间: 2014-05
• 期刊: HUMAN GENETICS
• 影响因子: 5.3
• 作者: Xiong, Xin;Xu, Chengqi;Zhang, Yuting;Li, Xiuchun;Wang, Binbin;Wang, Fan;Yang, Qin;Wang, Dan;Wang, Xiaojing;Li, Sisi;Chen, Shanshan;Zhao, Yuanyuan;Yin, Dan;Huang, Yufeng;Zhu, Xuan;Wang, Li;Wang, Longfei;Chang, Le;Xu, Chaoping;Li, Hui;Ke, Tie;Ren, Xiang;Wu, Yanxia;Zhang, Rongfeng;Wu, Tangchun;Xia, Yunlong;Yang, Yanzong;Ma, Xu;Tu, Xin;Wang, Qing K.
• 通讯作者: Wang, Qing K.

Regulation of CARD8 expression by ANRIL and association of CARD8 single nucleotide polymorphism rs2043211 (p.C10X) with ischemic stroke.

ANRIL 对 CARD8 表达的调节以及 CARD8 单核苷酸多态性 rs2043211 (p.C10X) 与缺血性中风的关联

• DOI: 10.1161/strokeaha.113.003393
• 发表时间: 2014-02
• 期刊: Stroke
• 影响因子: 8.3
• 作者: Bai Y;Nie S;Jiang G;Zhou Y;Zhou M;Zhao Y;Li S;Wang F;Lv Q;Huang Y;Yang Q;Li Q;Li Y;Xia Y;Liu Y;Liu J;Qian J;Li B;Wu G;Wu Y;Wang B;Cheng X;Yang Y;Ke T;Li H;Ren X;Ma X;Liao Y;Xu C;Tu X;Wang QK
• 通讯作者: Wang QK

A de novo mutation in NKX2.5 associated with atrial septal defects, ventricular noncompaction, syncope and sudden death.

• DOI: 10.1016/j.cca.2010.09.035
• 发表时间: 2011-01-14
• 期刊: Clinica chimica acta; international journal of clinical chemistry
• 作者: Ouyang P;Saarel E;Bai Y;Luo C;Lv Q;Xu Y;Wang F;Fan C;Younoszai A;Chen Q;Tu X;Wang QK
• 通讯作者: Wang QK