Comprehensive Epilepsy Center, New York University School of Medicine, New York, New York, United States of America.
Proteomics Laboratory, Division of Advanced Research Technologies, NYU School of Medicine, New York, New York, United States of America.
PLoS One. 2022 May 19;17(5):e0268597. doi: 10.1371/journal.pone.0268597. eCollection 2022.
Tuberous sclerosis complex (TSC) and some focal cortical dysplasias (FCDs) are associated with dysfunctional mTOR signaling, resulting in increased cell growth and ribosomal S6 protein phosphorylation (phospho-S6). mTOR inhibitors can reduce TSC tumor growth and seizure frequency, and preclinical FCD studies indicate seizure suppression. This pilot study evaluated safety of mTOR inhibitor everolimus in treatment resistant (failure of >2 anti-seizure medications) TSC and FCD patients undergoing surgical resection and to assess mTOR signaling and molecular pathways.
We evaluated everolimus in 14 treatment resistant epilepsy patients undergoing surgical resection (4.5 mg/m2 daily for 7 days; n = 4 Active, mean age 18.3 years, range 4-26; n = 10, Control, mean age 13.1, range 3-45). Everolimus was well tolerated. Mean plasma everolimus in Active participants were in target range (12.4 ng/ml). Brain phospho-S6 was similar in Active and Control participants with a lower trend in Active participants, with Ser235/236 1.19-fold (p = 0.67) and Ser240/244 1.15-fold lower (p = 0.66). Histologically, Ser235/236 was 1.56-fold (p = 0.37) and Ser240/244 was 5.55-fold lower (p = 0.22). Brain proteomics identified 11 proteins at <15% false discovery rate associated with coagulation system (p = 1.45x10-9) and acute phase response (p = 1.23x10-6) activation. A weighted gene correlation network analysis (WGCNA) of brain proteomics and phospho-S6 identified 5 significant modules. Higher phospho-S6 correlated negatively with cellular respiration and synaptic transmission and positively with organophosphate metabolic process, nuclear mRNA catabolic process, and neuron ensheathment. Brain metabolomics identified 14 increased features in Active participants, including N-acetylaspartylglutamic acid. Plasma proteomics and cytokine analyses revealed no differences.
Short-term everolimus before epilepsy surgery in TSC and FCD resulted in no adverse events and trending lower mTOR signaling (phospho-S6). Future studies should evaluate implications of our findings, including coagulation system activation and everolimus efficacy in FCD, in larger studies with long-term treatment to better understand molecular and clinical effects.
ClinicalTrials.gov NCT02451696.
结节性硬化症（TSC）和一些局灶性皮质发育不良（FCD）与功能失调的 mTOR 信号有关，导致细胞生长和核糖体 S6 蛋白磷酸化（磷酸化-S6）增加。mTOR 抑制剂可减少 TSC 肿瘤生长和癫痫发作频率，且 FCD 的临床前研究表明具有抑制癫痫作用。本研究旨在评估 mTOR 抑制剂依维莫司在接受手术切除的耐药性（抗癫痫药物治疗失败>2 种）TSC 和 FCD 患者中的安全性，并评估 mTOR 信号和分子途径。
我们评估了 14 名接受手术切除的耐药性癫痫患者（每天 4.5mg/m2，持续 7 天；n=4 名活性药物组，平均年龄 18.3 岁，范围 4-26；n=10 名对照组，平均年龄 13.1 岁，范围 3-45）使用依维莫司的情况。依维莫司的耐受性良好。活性药物组的平均血浆依维莫司浓度在目标范围内（12.4ng/ml）。活性药物组和对照组的脑磷酸化-S6 相似，活性药物组的趋势较低，Ser235/236 降低 1.19 倍（p=0.67），Ser240/244 降低 1.15 倍（p=0.66）。组织学上，Ser235/236 降低 1.56 倍（p=0.37），Ser240/244 降低 5.55 倍（p=0.22）。脑蛋白质组学鉴定出 11 个假发现率<15%的蛋白与凝血系统（p=1.45x10-9）和急性期反应（p=1.23x10-6）激活有关。脑蛋白质组学和磷酸化-S6 的加权基因相关性网络分析（WGCNA）鉴定出 5 个显著模块。较高的磷酸化-S6 与细胞呼吸和突触传递呈负相关，与有机磷代谢过程、核 mRNA 分解代谢过程和神经元鞘形成呈正相关。脑代谢组学在活性药物组中发现了 14 个增加的特征，包括 N-乙酰天冬氨酸谷氨酸。血浆蛋白质组学和细胞因子分析未发现差异。
TSC 和 FCD 患者在癫痫手术前短期使用依维莫司未发生不良事件，且 mTOR 信号（磷酸化-S6）呈下降趋势。未来的研究应评估我们的发现的意义，包括凝血系统的激活和依维莫司在 FCD 中的疗效，需要更大规模的长期治疗研究来更好地了解分子和临床效果。
ClinicalTrials.gov NCT02451696。
