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，以支持结节硬化症复杂癫痫的新疗法（TSC），该疾病包括早期癫痫，智力低下和自闭症。我们提出，氯化物共转运蛋白NKCC1的抑制剂Bumetanide将有效预防单独的TSC或与规范的GABA激动剂苯巴比妥或最近批准的抗惊厥药的抗癫痫发作。我们最近的出版物表明，NKCC1在人类TSC组织以及难治性癫痫，局灶性皮质发育不良IIB（FCD IIB）中过表达。 NKCC1增加会导致细胞内氯化物的增强，并且这种损害了GABA受体激活的抑制作用，从而导致它介导去极化和激发。我们最近生成了一种新型的TSC1CC Nestin-rtTA+ Tet-op-cre+突变小鼠模型，该模型是癫痫病，并显示了NKCC1的过表达和去极化GABA响应。 AIM 1将使用该小鼠模型单独或与苯巴比妥或vigabatrin在皮质切片中的体外和使用EEG记录在体内处理中的苯巴比妥或Vigabatrin的功效。 AIM 2将检查单独使用bumetanide并与苯巴比妥或vigabatrin结合使用的对脑切片中GABA反应的影响，该脑切片急性化，由人TSC和FCD IIB IIB手术活检组织制备。 拟议的工作代表了一种研究TSC中癫痫发生机制的新方法，并且有可能在TSC患者中产生一种基于新型机制的辅助机制抗惊厥策略。它还将为该机制提供原则证明，以及具有临床相关结果的TSC小鼠模型中的临床前功效。最后，我们的建议包括人类组织中的目标验证以及机制。这些是该化合物在TSC中进行首次人类试验的途径的关键要素，也可能延伸到FCD IIB。