中国癫痫患者三种HCN1变异的表型和功能变化特征

IF 2 4区医学 Q3 CLINICAL NEUROLOGY

Epilepsy Research Pub Date : 2025-02-07 DOI:10.1016/j.eplepsyres.2025.107521

Ziyao Han, Lingling Xie, Xiaorui Liu, Jiaxin Yang, Hanyu Luo, Ran Ding, Hengsheng Chen, Li Cheng, Zhixu Fang, Li Jiang

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引用次数: 0

摘要

目的评价中国癫痫患者HCN1基因变异位点的电生理特性，探讨基因型与表型之间的潜在关系。方法我们将3例HCN1变异体患者的临床严重程度与全细胞膜片钳测量的通道活性、Western blot检测的通道表达以及每种变异体损伤效应的生物信息学预测相关联。结果3例p.L400P、p.D534H和p.M243delinsTL变异患者表现出从轻度癫痫到重度癫痫性脑病的不同表型。变体L400P和D534H被所有生物信息学工具归类为致病性，变体M243delinsTL被MutationTaster归类为多态性。与野生型（WT）相比，L400P和D534H突变体的电流密度明显降低，而M243delinsTL突变体的电流密度与WT相似。与野生型相比，M243delinsTL突变体的半激活电压（V1/2）向超极化方向偏移。M243delinsTL突变体的激活斜率因子(k)显著低于WT， L400P突变体的激活时间常数显著高于WT。定量检测flag标记的HCN1通道显示，L400P变异的表达水平较wt显著降低。结论结合生物信息学工具和膜片钳技术确定变异位点的类型和位置，可以提高我们对HCN1变异相关癫痫临床表型的理解。

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Characterization of the phenotype and functional alternations of three HCN1 variants in Chinese epilepsy patients

Purpose

To evaluate the electrophysiological properties of three HCN1 variant sites found in Chinese epileptic patients and to explore the potential relationship between genotype and phenotype.

Methods

We correlated clinical severity of three patients with HCN1 variants with whole-cell patch-clamp measurements of channel activity, channel expression detected by Western blot, and bioinformatics prediction of the damaging effects of each variant.

Results

Three patients with the variants p.L400P, p.D534H and p.M243delinsTL, showed different phenotypes, ranging from mild epilepsy to severe epileptic encephalopathy. Variants L400P and D534H were classified as pathogenic by all bioinformatics tools, and variant M243delinsTL was classified as a polymorphism by MutationTaster. The L400P and D534H variants showed significantly reduced current compared with that of the wild-type (WT), while the current density of M243delinsTL was similar to WT. The half-activation voltage (V_1/2) of M243delinsTL variant was shifted in the hyperpolarizing direction when compared to the WT, and the slope factor (k) of activation of the M243delinsTL variant was significantly lower than that of the WT. The L400P variant was associated with a significantly higher activation time constant compared with that of the WT. In addition, quantitative detection of the FLAG-tagged HCN1 channel revealed that the expression level of the L400P variant was significantly decreased compared to WT.

Conclusions

Elucidation of the type and location of variant sites combined with the use of bioinformatics tools and patch-clamp techniques can improve our understanding of the clinical phenotype of epilepsy associated with HCN1 variants.

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来源期刊

Epilepsy Research 医学-临床神经学

CiteScore

0.10

自引率

4.50%

发文量

143

审稿时长

62 days

期刊介绍： Epilepsy Research provides for publication of high quality articles in both basic and clinical epilepsy research, with a special emphasis on translational research that ultimately relates to epilepsy as a human condition. The journal is intended to provide a forum for reporting the best and most rigorous epilepsy research from all disciplines ranging from biophysics and molecular biology to epidemiological and psychosocial research. As such the journal will publish original papers relevant to epilepsy from any scientific discipline and also studies of a multidisciplinary nature. Clinical and experimental research papers adopting fresh conceptual approaches to the study of epilepsy and its treatment are encouraged. The overriding criteria for publication are novelty, significant clinical or experimental relevance, and interest to a multidisciplinary audience in the broad arena of epilepsy. Review articles focused on any topic of epilepsy research will also be considered, but only if they present an exceptionally clear synthesis of current knowledge and future directions of a research area, based on a critical assessment of the available data or on hypotheses that are likely to stimulate more critical thinking and further advances in an area of epilepsy research.