A murine model for human factor H R1210C mutation-related diseases

人类因子HR1210C突变相关疾病的小鼠模型

• 批准号: 8652434
• 负责人: Wenchao Song
• 金额: $ 20万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-16 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8652434
• 关键词: Affect; Affinity; Age related macular degeneration; Alternative Complement Pathway; Amino Acids; Animal Model; Arginine; Binding; Blood Proteins; C-terminal; Cattle; Cells; Complement; Complement 3b; Complement 3d; Complement Factor H; Cysteine; Defect; Deposition; Development; Dialysis procedure; Disease; Endothelial Cells; Family suidae; Figs - dietary; Gene Targeting; Genetic; Grant; Hemolytic-Uremic Syndrome; Heparin; Human; Human Factor H; In Vitro; Individual; Infection Control; Kidney Diseases; Knowledge; Light; Modeling; Mus; Mutagenesis; Mutant Strains Mice; Mutation; N-terminal; Pathogenesis; Pathology; Patients; Plasma; Positioning Attribute; Rattus; Retinal Degeneration; Risk; Structure; Surface; Sushi Domain; Testing; Tissues; Transplantation; Variant; human disease; inhibitor/antagonist; interest; mouse model; mutant; public health relevance

DESCRIPTION (provided by applicant): Factor H (fH) is a plasma inhibitor of the alternative pathway (AP) complement. It is composed of 20 short consensus repeat (SCR) domains. The complement regulatory function of fH is located in the N-terminal SCR1-4, whereas the two C-terminal SCRs of fH (SCR 19 and 20) are critical for interacting with surface deposited C3b/C3d in the context of host cell-specific polyanionic constituents. Mutations in SCR19-20 of human fH impair its interaction with host cells and predispose to the development of atypical hemolytic uremic syndrome (aHUS). Among the various SCR19-20 mutations of fH that are found in aHUS patients, the R1210C mutation is of particular interest as a recent genetic study has identified this mutation to be also a high-penetrant mutation for the development of age-related macular degeneration (AMD). The R1210C variant of human fH showed reduced binding to C3b/C3d, heparin and endothelial cells, yet R1210 is not conserved across species (D1210 in mouse, G1210 in rat, P1210 in cows and pigs). We hypothesize that the change of amino acid at position 1210 of human fH to cysteine, rather than the simple loss of arginine, is critical for conferring the risk of aHUS and AMD. In this exploratory grant, we propose two specific aims to test this hypothesis. Specific Aim 1. We will perform in vitro mutagenesis studies of human and mouse fH at residue 1210 and test the prediction that a mouse D1210C mutant will, like human R1210C mutant, have impaired binding to C3b, heparin and endothelial cells. On the other hand, we predict that mutations of R1210 in human fH to R1210D, R1210G and R1210P (to mouse, rat and cow/pig residue, respectively), and mutations of D1210 in mouse fH to D1210R, D1210G and D1210P (to human, rat, and cow/pig residue, respectively), are tolerated and produce no functional consequences for C3b-, heparin- and endothelial cell-binding. Specific Aim 2. We will generate by gene targeting a D1210C mutant mouse and determine if this mouse develops aHUS and/or retinal degeneration resembling human AMD. These studies will shed new light on structure/function knowledge of the fH C-terminal domain and establish how specific mutations in this domain may differentially affect fH function in different tissues and species. Furthermore, they will help us understand the pathogenesis of two fH-related human diseases and provide a much- needed mouse model for testing anti-complement therapies for these pathologies.

描述（由申请人提供）：因子H（FH）是替代途径（AP）补体的血浆抑制剂。它由20个短共识重复（SCR）域组成。 FH的补体调节功能位于N端SCR1-4中，而FH的两个C末端SCR（SCR 19和20）对于与表面沉积的C3B/C3D相互作用至关重要。聚支号成分。 SCR19-20的人类FH的突变损害了其与宿主细胞的相互作用，并且易感非典型溶血性尿毒症综合征（AHUS）的发展。在AHUS患者中发现的各种SCR19-20 FH突变中，R1210C突变特别令人感兴趣，因为最近的遗传研究已经确定该突变也是一种高渗透剂突变，用于发展与年龄相关的黄斑变性（ AMD）。人类FH的R1210C变体显示与C3B/C3D，肝素和内皮细胞的结合降低，但R1210并未在物种之间保守（小鼠中的D1210，大鼠的G1210，大鼠的G1210，牛和猪中的P1210）。我们假设人类FH位置1210的氨基酸变为半胱氨酸，而不是简单的精氨酸损失，对于赋予AHUS和AMD的风险至关重要。在这项探索性赠款中，我们提出了两个特定的目的来检验这一假设。具体目的1。我们将对残基1210对人和小鼠FH进行体外诱变研究，并测试小鼠D1210C突变体将像人R1210C突变体一样的预测，也损害了与C3B，肝素和内皮细胞的结合。另一方面，我们预测，在人FH到R1210D，R1210G和R1210P中R1210的突变（分别为鼠标，大鼠和牛/猪残留），而鼠标FH至D1210R中D1210的突变是D1210R，D1210G，D1210G和D1210P（对人类的分别对C3B，肝素和内皮细胞结合产生功能后果，分别是大鼠和牛/猪残留物。具体目标2。我们将通过靶向D1210C突变小鼠的基因生成，并确定该小鼠是否形成了类似于人类AMD的Ahus和/或视网膜变性。这些研究将为FH C末端结构域的结构/功能知识提供新的启示，并确定该域中的特定突变如何在不同组织和物种中差异化FH功能。此外，它们将帮助我们了解两种与FH相关的人类疾病的发病机理，并提供了急需的小鼠模型，用于测试这些病理学的抗补充疗法。