The role of beta-catenin in the pathophysiology of infantile spasms

β-连环蛋白在婴儿痉挛症病理生理学中的作用

• 批准号: 10057263
• 负责人: Chris G Dulla
• 金额: $ 46.05万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10057263
• 关键词: APC gene; Address; Adhesions; Adult; Adverse effects; Age; Age-Months; Alleles; Animal Model; Attenuated; Behavior; Behavioral; Blood; Brain; Brain region; Breeding; Caring; Child; Clinical; Cognitive deficits; Communication; Complex; Data; Databases; Development; Developmental Delay Disorders; Dose; Drug Kinetics; Electroencephalography; Enterobacteria phage P1 Cre recombinase; Epilepsy; Excitatory Synapse; Exhibits; Family; Female; Frequencies; Functional disorder; Future; Gene Family; Generations; Genes; Genetic; Genetic Transcription; Goals; Head and neck structure; Hippocampus (Brain); Human; Human Characteristics; Human Genetics; Impaired cognition; Infantile spasms; Knockout Mice; Lead; Life; Limb structure; Link; LoxP-flanked allele; Mediating; Modeling; Molecular; Mus; Mutation; N-Cadherin; Neonatal; Neurons; Pathway interactions; Pharmaceutical Preparations; Pharmacodynamics; Pharmacology; Pharmacotherapy; Phenocopy; Phenotype; Pre-Clinical Model; Predisposition; Prevention; Regimen; Rodent Model; Role; Seizures; Signal Pathway; Signal Transduction; Spasm; Synapses; Syndrome; System; Tankyrase; Testing; Therapeutic; Treatment Protocols; Variant; Vertebral column; WNT Signaling Pathway; adverse outcome; base; beta catenin; brain cell; calmodulin-dependent protein kinase II; cell type; childhood epilepsy; conditional knockout; curative treatments; density; developmental disease; epileptic encephalopathies; excitatory neuron; experience; experimental study; hippocampal pyramidal neuron; improved outcome; in vivo; inhibitor/antagonist; innovation; insight; knockout animal; male; mouse model; neonatal brain development; neonate; new therapeutic target; novel; offspring; overexpression; pharmacodynamic model; post stroke; postnatal; pre-clinical; promoter; risk variant; side effect; therapeutic target; treatment strategy

Project summary Infantile spasms (IS, also known as West Syndrome) is a catastrophic childhood epilepsy syndrome characterized by spasms which progress into seizures later in life. Spasms are typified by spontaneous flexion/extension of the head, neck, and limbs and occur first between 4 and 8 months of age. The current treatment options for IS are often ineffective and are associated with significant side effects. Therefore, novel treatment strategies are essential. One limiting factor in identifying new treatment approaches is a paucity of pre-clinical animal models. We have identified and characterized a novel rodent model with many phenotypic characteristics of human IS. The model was generated by breeding male mice containing a floxed version of the Adenomatous polyposis coli (APC) gene with female mice expressing the Cre-recombinase gene under the control of the Ca2+/calmodulin-dependent protein kinase II alpha (CaMKIIa) promoter. The offspring of this cross, which lack APC in CaMKIIa-positive neurons, are known as APC conditional knockouts (APC cKOs). APC cKO animals have been shown to have increased excitatory synaptic communication and an increased density of excitatory spines on hippocampal CA1 pyramidal neurons. APC is the main inhibitory regulator of a large signaling pathway known as the β-catenin/Wnt pathway. APC is part of the β-catenin destruction complex, targeting β-catenin for degradation. When APC is lost, β-catenin levels rise and 1) increase transcription of a large family of genes, and 2) increase the stability of excitatory synapses. We began by examining APC cKO animals for phenotypes consistent with human IS. We found that they exhibit spontaneous behavioral spasms at postnatal days 8-11, they have an ictal EEG correlate of spasm behavior similar to human ictal activity in IS, and as adults, they have spontaneous electrographic and behavioral seizures. Interestingly, APC heterozygous mutations in humans are linked to both developmental and seizure disorders. Furthermore, many of the genes linked to IS are either part of the β-catenin/Wnt pathway or are reciprocally regulated by it. In this proposal, we will specifically examine the role of β-catenin in the pathophysiology of infantile spasms. We will examine the effects of increasing β-catenin (by deleting APC and independently of APC) on spasm behavior, seizures, and electrographic brain activity. Next, we will perform careful pharmacokinetic, pharmacodynamic, and adverse effect analysis of manipulating β-catenin during development with a drug called G007-LK. Lastly, we will determine if restoring β-catenin levels to normal attenuates spasms and seizures later in life. This proposal will address the role of β-catenin in the pathophysiology of spasms, provide a new mouse model for pre-clinical analysis, and introduce a large set of new potential therapeutic targets for the treatment of IS.

项目摘要 婴儿痉挛（IS也称为西综合征）是一种灾难性的儿童癫痫综合症 痉挛的特征是痉挛，后来癫痫发作。痉挛是自发的 头部，颈部和四肢的屈曲/伸展，首先发生在4至8个月大之间。电流 IS的治疗选择通常无效，并且与重大副作用有关。因此，新颖 治疗策略至关重要。识别新治疗方法的一个限制因素是很少 临床前动物模型。我们已经确定并表征了具有许多表型的新型啮齿动物模型 人的特征是。该模型是通过繁殖的雄性小鼠而产生的 腺瘤性息肉病大肠杆菌（APC）基因，雌性小鼠在该基因下表达CRE聚合酶基因 控制Ca2+/钙调蛋白依赖性蛋白激酶IIα（CAMKIIA）启动子的控制。这个后代 Cross在CAMKIIA阳性神经元中缺乏APC，被称为APC条件敲除（APC CKO）。 APC CKO动物已显示出增加兴奋性突触通信的增加，并且增加了 海马CA1锥体神经元上兴奋性棘的密度。 APC是A的主要抑制性调节剂 大型信号通路称为β-catenin/Wnt途径。 APC是β-catenin破坏的一部分 复合物，靶向β-catenin进行降解。当APC丢失时，β-catenin水平上升，1）增加 大型基因家族的转录，以及2）提高兴奋性突触的稳定性。我们开始 检查与人为一致的表型的APC CKO动物。我们发现他们展示了 在产后第8-11天的赞助行为痉挛，它们具有痉挛行为的发作性EEG相关性 与IS中的人类发射活性类似，作为成年人，它们具有赞助电报和行为。 癫痫发作。有趣的是，人类的APC杂合突变与发育和癫痫发作有关 疾病。此外，与IS相关的许多基因要么是β-catenin/wnt途径的一部分，要么是 受其相互监管。在此提案中，我们将专门研究β-catenin在 婴儿痉挛的病理生理。我们将检查增加β-catenin的影响（通过删除APC和 独立于APC）关于痉挛行为，癫痫发作和电图脑活动。接下来，我们将表演 仔细的药代动力学，药效和不良反应分析在操纵β-catenin期间 使用称为G007-LK的药物开发。最后，我们将确定是否将β-catenin水平恢复到正常 减轻痉挛和癫痫发作的后期。该提案将解决β-catenin在 痉挛的病理生理学，为临床前分析提供了新的鼠标模型，并引入了大量 用于治疗IS的新潜在治疗靶标。