Li Yibo, Sadri Zahra, Blandin Katherine J, Narvaiz David A, Aryal Uma K, Lugo Joaquin N, Poolos Nicholas P, Brewster Amy L
Department of Biological Sciences, Southern Methodist University, Dallas, TX, United States of America.
Department of Psychology and Neuroscience, Baylor University, Waco, TX, United States of America.
Exp Neurol. 2025 Jun 24;392:115361. doi: 10.1016/j.expneurol.2025.115361.
Epilepsy presents significant sex-based disparities in prevalence and manifestation. Epidemiological studies reveal that epilepsy is more prevalent in males, with lesional types being more common, whereas idiopathic generalized epilepsies are more frequently observed in females. These differences stress the importance of considering sex-specific factors in epilepsy diagnosis, treatment, and mechanistic research using preclinical models. To elucidate potential molecular differences that could explain these disparities and inform personalized treatment strategies, we conducted a proteomic analysis of epileptic brain tissues from both an experimental mouse model of genetic epilepsy and humans with drug-resistant epilepsy (DRE).
We employed mass spectrometry-based proteomic analysis on brain tissues from DRE patients and the Pten knockout (KO) mouse model of genetic epilepsy with focal cortical dysplasia. Mouse samples included hippocampi from adult wild-type (WT) and Pten KO mice (4-5 per group and sex). Human samples included the temporal cortex from 12 DRE adult patients (7 males, 5 females) and 5 non-epileptic (NE) controls (2 males, 3 females). Brain biopsies were collected with patients' informed consent under approved IRB protocols (Indiana University Health Biorepository). Proteomic profiles were analyzed using principal component analysis (PCA) along with volcano plots to identify significant changes in protein expression. The enrichment analysis of differentially expressed proteins was conducted by Gene Ontology (GO) and Kyoto Encyclopedia of Gene and Genomes (KEGG) pathway.
PCA revealed distinct clustering of brain proteomes between epilepsy and control cases in both human and mice, with 390 proteins showing significant differences in human and 437 proteins in mouse samples. These proteins are primarily associated with ion channels, synaptic processes, and neuronal energy regulation. In the mouse model, males have more pronounced proteomic changes than females, with enrichment in metabolic pathways and VEGF signaling pathway, indicating a more severe vascular permeability impairment in males. In human DRE cases, 118 proteins were significantly changed by comparing epileptic females to males. Pathway analysis revealed changes in metabolic pathways and the HIF-1 signaling pathway, indicating that altered neuronal activity and inflammation may lead to increased oxygen consumption.
These findings highlight differences between epilepsy and control brain samples in both humans and mice. Sex-specific analysis revealed distinct pathway enrichments between females and males, with males exhibiting a broader range of proteomic alterations. While these observations suggest potential sex-related differences in proteomic profiles, larger studies are needed to further validate these patterns. This exploratory work provides initial insights into possible underlying mechanisms of epilepsy and suggests that sex may be an important consideration in future epilepsy studies, though more comprehensive studies are required to establish therapeutic interventions.
癫痫在患病率和表现方面存在显著的性别差异。流行病学研究表明，癫痫在男性中更为普遍，病灶性类型更为常见，而特发性全身性癫痫在女性中更为常见。这些差异凸显了在癫痫诊断、治疗以及使用临床前模型进行机制研究时考虑性别特异性因素的重要性。为了阐明可能解释这些差异并为个性化治疗策略提供依据的潜在分子差异，我们对遗传性癫痫实验小鼠模型和耐药性癫痫（DRE）患者的癫痫脑组织进行了蛋白质组学分析。
我们对DRE患者的脑组织以及具有局灶性皮质发育异常的遗传性癫痫Pten基因敲除（KO）小鼠模型进行了基于质谱的蛋白质组学分析。小鼠样本包括成年野生型（WT）和Pten KO小鼠的海马体（每组和每种性别各4 - 5只）。人类样本包括12名成年DRE患者（7名男性，5名女性）和5名非癫痫（NE）对照者（2名男性，3名女性）的颞叶皮质。在获得患者知情同意的情况下，根据批准的机构审查委员会方案（印第安纳大学健康生物样本库）收集脑活检样本。使用主成分分析（PCA）和火山图分析蛋白质组学图谱，以识别蛋白质表达的显著变化。通过基因本体论（GO）和京都基因与基因组百科全书（KEGG）通路对差异表达蛋白质进行富集分析。
PCA显示，人类和小鼠的癫痫与对照病例之间的脑蛋白质组存在明显聚类，人类中有390种蛋白质显示出显著差异，小鼠样本中有437种蛋白质显示出显著差异。这些蛋白质主要与离子通道、突触过程和神经元能量调节有关。在小鼠模型中，雄性的蛋白质组变化比雌性更明显，在代谢途径和VEGF信号通路中富集，表明雄性的血管通透性损害更严重。在人类DRE病例中，将癫痫女性与男性进行比较时，有118种蛋白质发生了显著变化。通路分析显示代谢途径和HIF - 1信号通路发生了变化，表明神经元活动和炎症的改变可能导致氧消耗增加。
这些发现突出了人类和小鼠癫痫与对照脑样本之间的差异。性别特异性分析显示，雌性和雄性之间存在不同的通路富集，雄性表现出更广泛的蛋白质组改变。虽然这些观察结果表明蛋白质组图谱可能存在与性别相关的差异，但需要更大规模的研究来进一步验证这些模式。这项探索性工作为癫痫可能的潜在机制提供了初步见解，并表明性别可能是未来癫痫研究中的一个重要考虑因素，不过需要更全面的研究来建立治疗干预措施。
