The NKCC1 inhibitor bumetanide as a novel therapy in TSC

NKCC1 抑制剂布美他尼作为 TSC 的新型疗法

• 批准号: 8554384
• 负责人: Frances E Jensen
• 金额: $ 18.79万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8554384
• 关键词: Address; Adjuvant; Affect; Agonist; Animal Model; Animals; Anticonvulsants; Antiepileptic Agents; Applications Grants; Area; Autistic Disorder; Benchmarking; Biopsy; Brain; Brain Pathology; Bumetanide; Cells; Child; Chloride Ion; Chlorides; Clinical; Cortical Dysplasia; Data; Disease; Diuretics; Drug Kinetics; Electroencephalography; Elements; Epilepsy; Epileptogenesis; Equilibrium; Excision; Exhibits; FDA approved; GABA Agonists; GABA Receptor; Genes; Giant Cells; Gramicidin; Health; Henle' s loop; Hereditary Disease; Human; In Vitro; Incidence; Intellectual functioning disability; Left; Life; Measures; Mediating; Mental Retardation; Modeling; Morphology; Mus; Neonatal; Neuroglia; Neurologic; Neurons; New Agents; Operative Surgical Procedures; Outcome; Partial Epilepsies; Pathway interactions; Patients; Pharmaceutical Preparations; Phase I/II Trial; Phenobarbital; Population; Prevention; Publications; Rattus; Receptor Activation; Refractory; Relative (related person); Research; Resected; Resistance; Role; Safety; Sampling; Seizures; Sirolimus; Slice; Synapses; Techniques; Testing; Tetanus Helper Peptide; Text; Tissues; Transgenic Mice; Tuberous sclerosis protein complex; Up-Regulation; Validation; Vigabatrin; Wild Type Mouse; Work; base; clinically relevant; gamma-Aminobutyric Acid; hippocampal pyramidal neuron; human FRAP1 protein; human tissue; improved; in vivo; inhibitor/antagonist; innovation; mTOR Inhibitor; mouse model; mutant mouse model; neonate; nestin protein; new technology; novel; novel strategies; overexpression; pre-clinical; preclinical efficacy; prevent; receptor; response; sodium-potassium-chloride cotransporter 1 protein; therapeutic target; treatment strategy

DESCRIPTION (provided by applicant): This proposal aims to provide preclinical data in support of a novel therapy for epilepsy in Tuberous sclerosis complex (TSC), a disorder that includes early life epilepsy, mental retardation and autism. We propose that bumetanide, an inhibitor of the chloride cotransporter NKCC1, will be effective for the prevention of seizures in TSC alone or in combination with a canonical GABA agonist phenobarbital or the recently approved anticonvulsant vigabatrin. Our recent publication shows that NKCC1 is overexpressed in human TSC tissue, as well as in another cause of refractory epilepsy, focal cortical dysplasia Type IIb (FCD IIb). Increased NKCC1 causes enhanced intracellular chloride, and this impairs inhibitory actions of GABA receptor activation by causing it to mediate depolarization and excitation. We have recently generated a novel Tsc1cc Nestin-rtTA+ tet-OP-cre+ mutant mouse model that is epileptic and shows NKCC1 overexpression and depolarizing GABA responses. Aim 1 will use this mouse model to further text the efficacy of bumetanide alone or in combination with phenobarbital or vigabatrin in cortical slices in vitro and by treatment in vivo using EEG recordings. Aim 2 will examine the effects of bumetanide alone and in combination with phenobarbital or vigabatrin on GABA responses in brain slices acutely prepared from human TSC and FCD IIb surgical biopsy tissue. The proposed work represents a new approach in studying the mechanisms of epileptogenesis in TSC and has the potential to generate an adjuvant novel mechanism-based anticonvulsant strategy for seizure control in TSC patients. It will also provide proof-of-principle for this mechanism, as well as preclinical efficacy in a TSC mouse model with clinically relevant outcomes. Finally our proposal includes target validation as well as mechanism in human tissue. These are critical elements of a path to first-in-human trials for this compound in TSC, and may also extend to FCD IIb.

描述（由申请人提供）：该提案旨在提供临床前数据，支持结节性硬化症（TSC）癫痫的新疗法，TSC 是一种包括早期癫痫、智力低下和自闭症的疾病。我们建议布美他尼（氯协同转运蛋白 NKCC1 的抑制剂）单独使用或与经典 GABA 激动剂苯巴比妥或最近批准的抗惊厥药氨己烯酸联合使用可有效预防 TSC 癫痫发作。我们最近发表的文章表明，NKCC1 在人类 TSC 组织以及难治性癫痫的另一种原因——局灶性皮质发育不良 IIb 型 (FCD IIb) 中过度表达。 NKCC1 增加会导致细胞内氯离子增加，从而通过介导去极化和兴奋来损害 GABA 受体激活的抑制作用。我们最近生成了一种新型 Tsc1cc Nestin-rtTA+ tet-OP-cre+ 突变小鼠模型，该模型患有癫痫，并显示 NKCC1 过度表达和去极化 GABA 反应。目标 1 将使用该小鼠模型进一步证明布美他尼单独使用或与苯巴比妥或氨己烯酸联合使用在体外皮质切片中的功效以及通过脑电图记录进行体内治疗。目标 2 将检查布美他尼单独使用以及与苯巴比妥或氨己烯酸联合使用对由人类 TSC 和 FCD IIb 手术活检组织急性制备的脑切片中 GABA 反应的影响。 这项工作代表了研究 TSC 癫痫发生机制的新方法，并有可能为 TSC 患者的癫痫发作控制产生一种基于辅助新机制的抗惊厥策略。它还将为该机制提供原理验证，以及 TSC 小鼠模型的临床前疗效和临床相关结果。最后，我们的建议包括目标验证以及人体组织中的机制。这些是该化合物在 TSC 中进行首次人体试验的关键要素，也可能扩展到 FCD IIb。