Biomedical Laboratory Science, Department of Technology, Faculty of Health and Technology, University College Copenhagen, Sigurdsgade 26, 1St, 2200, Copenhagen, Denmark.
Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, 2100, Copenhagen, Denmark.
Mol Neurobiol. 2023 Oct;60(10):5755-5769. doi: 10.1007/s12035-023-03426-4. Epub 2023 Jun 21.
The purpose of this study was to identify and validate new putative lead drug targets in drug-resistant mesial temporal lobe epilepsy (mTLE) starting from differentially expressed genes (DEGs) previously identified in mTLE in humans by transcriptome analysis. We identified consensus DEGs among two independent mTLE transcriptome datasets and assigned them status as "lead target" if they (1) were involved in neuronal excitability, (2) were new in mTLE, and (3) were druggable. For this, we created a consensus DEG network in STRING and annotated it with information from the DISEASES database and the Target Central Resource Database (TCRD). Next, we attempted to validate lead targets using qPCR, immunohistochemistry, and Western blot on hippocampal and temporal lobe neocortical tissue from mTLE patients and non-epilepsy controls, respectively. Here we created a robust, unbiased list of 113 consensus DEGs starting from two lists of 3040 and 5523 mTLE significant DEGs, respectively, and identified five lead targets. Next, we showed that CACNB3, a voltage-gated Ca channel subunit, was significantly regulated in mTLE at both mRNA and protein level. Considering the key role of Ca currents in regulating neuronal excitability, this suggested a role for CACNB3 in seizure generation. This is the first time changes in CACNB3 expression have been associated with drug-resistant epilepsy in humans, and since efficient therapeutic strategies for the treatment of drug-resistant mTLE are lacking, our finding might represent a step toward designing such new treatment strategies.
本研究旨在从先前通过转录组分析在人类内侧颞叶癫痫（mTLE）中鉴定的差异表达基因（DEGs）出发，确定和验证耐药性 mTLE 中的新假定的潜在药物靶标。我们在两个独立的 mTLE 转录组数据集之间鉴定了共识 DEGs，如果它们（1）参与神经元兴奋性，（2）在 mTLE 中是新的，并且（3）可药用，则将其指定为“潜在靶标”。为此，我们在 STRING 中创建了一个共识 DEG 网络，并使用 DISEASES 数据库和 Target Central Resource Database（TCRD）中的信息对其进行了注释。接下来，我们分别使用 qPCR、免疫组织化学和 Western blot 尝试在 mTLE 患者的海马和颞叶新皮质组织以及非癫痫对照中验证潜在靶标。在这里，我们从分别包含 3040 和 5523 个 mTLE 显著 DEGs 的两个列表中创建了一个稳健、无偏的 113 个共识 DEG 列表，并确定了五个潜在靶标。接下来，我们表明 CACNB3（电压门控钙通道亚基）在 mTLE 中在 mRNA 和蛋白水平均受到显著调控。考虑到 Ca 电流在调节神经元兴奋性中的关键作用，这表明 CACNB3 在癫痫发作的产生中起作用。这是 CACNB3 表达变化与人类耐药性癫痫相关的首次报道，由于缺乏针对耐药性 mTLE 的有效治疗策略，我们的发现可能代表朝着设计此类新治疗策略迈出了一步。
