Functional Characterization of Rare Genetic Variants in the N-Terminus of Complement Factor H in aHUS, C3G, and AMD.
Functional Characterization of Rare Genetic Variants in the N-Terminus of Complement Factor H in aHUS, C3G, and AMD.
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DOI：
10.3389/fimmu.2020.602284
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发表时间：
2020
影响因子：
7.3
通讯作者：
Kavanagh D
中科院分区：
文献类型：
--
作者：
Wong EKS;Hallam TM;Brocklebank V;Walsh PR;Smith-Jackson K;Shuttleworth VG;Cox TE;Anderson HE;Barlow PN;Marchbank KJ;Harris CL;Kavanagh D
膜增生性肾小球肾炎（MPGN）、C3肾小球病（C3 G）、非典型溶血性尿毒症综合征（阿胡斯）和年龄相关性黄斑变性（AMD）都与补体旁路途径（AP）功能障碍密切相关。MPGN、C3 G、阿胡斯和AMD患者中有很大一部分携带CFH基因的罕见遗传变异，这些变异导致H因子（FH）蛋白（AP的重要调节因子）的功能或数量缺陷。对CFH中罕见遗传变异的有害性进行计算机模拟分析并不可靠，仔细的生化评估仍然是金标准。在患有AP的这些疾病的患者中鉴定了CFH中不确定意义的六个N-末端变体，并选择用于分析。在FH CCP 1-4的设置中在巴斯德毕赤酵母中产生变体，通过镍亲和层析和尺寸排阻纯化，并通过表面等离子体共振和溶血测定以及在流体相中的辅因子测定表征。一个单一的变体，Q81 P表现出与C3 b结合的严重损失，随之而来的是辅因子和衰变加速活性的损失。另外2种变体G69 E和D130 N仅表现出细微的缺陷，可以想象随着时间的推移，这些缺陷会导致更慢性的AP疾病如C3 G和AMD的疾病进展。在变体S159 N、A161 S和M162 V中，任何功能缺陷都低于实验测定可靠检测的能力。这项研究进一步强调了在将功能后果分配给可能改变患者临床决策的罕见遗传变异时，仔细的生化评估的重要性。
Membranoproliferative glomerulonephritis (MPGN), C3 glomerulopathy (C3G), atypical haemolytic uraemic syndrome (aHUS) and age-related macular degeneration (AMD) have all been strongly linked with dysfunction of the alternative pathway (AP) of complement. A significant proportion of individuals with MPGN, C3G, aHUS and AMD carry rare genetic variants in the CFH gene that cause functional or quantitative deficiencies in the factor H (FH) protein, an important regulator of the AP. In silico analysis of the deleteriousness of rare genetic variants in CFH is not reliable and careful biochemical assessment remains the gold standard. Six N-terminal variants of uncertain significance in CFH were identified in patients with these diseases of the AP and selected for analysis. The variants were produced in Pichia Pastoris in the setting of FH CCPs 1–4, purified by nickel affinity chromatography and size exclusion and characterized by surface plasmon resonance and haemolytic assays as well as by cofactor assays in the fluid phase. A single variant, Q81P demonstrated a profound loss of binding to C3b with consequent loss of cofactor and decay accelerating activity. A further 2 variants, G69E and D130N, demonstrated only subtle defects which could conceivably over time lead to disease progression of more chronic AP diseases such as C3G and AMD. In the variants S159N, A161S, and M162V any functional defect was below the capacity of the experimental assays to reliably detect. This study further underlines the importance of careful biochemical assessment when assigning functional consequences to rare genetic variants that may alter clinical decisions for patients.
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DOI：
10.4049/jimmunol.0804031
发表时间：
2009-06-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Ferreira VP;Herbert AP;Cortés C;McKee KA;Blaum BS;Esswein ST;Uhrín D;Barlow PN;Pangburn MK;Kavanagh D
通讯作者：
Kavanagh D
DOI：
10.1073/pnas.0501536102
发表时间：
2005-05-17
影响因子：
11.1
作者：
Hageman, GS;Anderson, DH;Allikmets, R
通讯作者：
Allikmets, R
影响因子：
13.6
作者：
Recalde, Sergio;Tortajada, Agustin;Rodriguez de Cordoba, Santiago
通讯作者：
Rodriguez de Cordoba, Santiago
影响因子：
3.5
作者：
Tortajada A;Montes T;Martínez-Barricarte R;Morgan BP;Harris CL;de Córdoba SR
通讯作者：
de Córdoba SR
DOI：
10.1074/jbc.m110.211839
发表时间：
2011-04-01
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Pechtl IC;Kavanagh D;McIntosh N;Harris CL;Barlow PN
通讯作者：
Barlow PN