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: 9293864
• 负责人: Chris G Dulla
• 金额: $ 49.45万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9293864
• 关键词: APC gene; Address; Adenomatous Polyposis Coli; Adhesions; Adult; Adverse effects; Aftercare; Age; Age-Months; Alleles; Animal Model; Animals; Attenuated; Behavior; Behavioral; Blood; Brain; Brain region; Breeding; Caring; Child; Childhood; Clinical; Cognition; Communication; Complex; Data; Development; Developmental Delay Disorders; Dose; Drug Kinetics; Early treatment; Electroencephalography; Epilepsy; Excitatory Synapse; Exhibits; Family; Female; Functional disorder; Gene Family; Genes; Genetic Models; Genetic Transcription; Head and neck structure; Hippocampus (Brain); Human; Human Characteristics; Impaired cognition; Infantile spasms; Knock-out; Lead; Life; Limb structure; Link; LoxP-flanked allele; Mediating; Modeling; Molecular; Mus; Mutation; N-Cadherin; Neonatal; Neurologic; Neurons; Pathology; Pathway interactions; Pharmaceutical Preparations; Pharmacodynamics; Pharmacotherapy; Phenocopy; Phenotype; Predisposition; Prevention; Property; Regimen; Rodent Model; Role; Seizures; Signal Pathway; Signal Transduction; Spasm; Synapses; Syndrome; Tankyrase; Testing; Therapeutic; Vertebral column; adverse outcome; beta catenin; brain cell; calmodulin-dependent protein kinase II; cell type; density; developmental disease; excitatory neuron; genetic linkage; hippocampal pyramidal neuron; in vivo; inhibitor/antagonist; insight; male; mouse model; neonatal brain; neonate; new therapeutic target; novel; offspring; overexpression; post stroke; pre-clinical; prevent; promoter; recombinase; research study; therapeutic target; therapy design; 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 between 4-8 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 ß-catenin/Wnt pathway. APC is part of the ß-catenin destruction complex, targeting ß-catenin for degradation. When APC is lost, ß-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 from post-natal day 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 are linked to both developmental and seizure disorders. Furthermore, many of the genes linked to IS are either part of the ß-catenin/Wnt pathway or are reciprocally regulated by it. In this proposal we will specifically examine the role of ß-catenin in the pathophysiology of infantile spasms. We will examine the effects of increasing ß-catenin by deleting APC, and independently of APC, on spasms behavior, seizure, and electrographic brain activity. Next, we will perform careful pharmacokinetic, pharmacodynamic, and adverse effect analysis of manipulating ß-catenin during development. Lastly, we will determine if restoring ß-catenin levels to normal attenuates spasms and seizures later in life. This proposal will address the role of ß-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.

项目概要 婴儿痉挛症（IS，也称为韦斯特综合症）是一种灾难性的儿童癫痫综合症 其特征是痉挛，并在以后的生活中发展为癫痫发作。痉挛的典型特征是自发性痉挛 头部、颈部和四肢的屈曲/伸展发生在 4-8 个月大时。目前的治疗 IS 的选择通常无效，并且会产生严重的副作用。因此，新的治疗 战略是必不可少的。识别新治疗方法的一个限制因素是缺乏临床前研究 动物模型。我们已经鉴定并表征了一种具有多种表型的新型啮齿动物模型 人类IS的特征。该模型是通过饲养含有 floxed 版本的雄性小鼠而产生的 腺瘤性结肠息肉病 (APC) 基因与表达 Cre 重组酶基因的雌性小鼠 Ca2+/钙调蛋白依赖性蛋白激酶 II α (CaMKIIa) 启动子的控制。这个的后代 CaMKIIa 阳性神经元中缺乏 APC，称为 APC 条件敲除 (APC cKO)。 APC cKO 动物已被证明具有增加的兴奋性突触通讯和增加的 海马 CA1 锥体神经元兴奋性棘的密度。 APC是主要的抑制性调节因子 称为 ß-catenin/Wnt 途径的大信号传导途径。 APC 是 β-连环蛋白破坏的一部分 复合物，针对β-连环蛋白进行降解。当 APC 丢失时，β-连环蛋白水平升高并且 1) 增加 一个大基因家族的转录，2) 增加兴奋性突触的稳定性。我们开始于 检查 APC cKO 动物的表型是否与人类 IS 一致。我们发现他们展示 产后第 8-11 天出现自发行为痉挛，发作期脑电图与痉挛行为相关 与 IS 中的人类发作活动相似，并且作为成年人，他们有自发的电图和行为 癫痫发作。有趣的是，APC 杂合突变与发育障碍和癫痫发作有关。 此外，许多与 IS 相关的基因要么是 ß-catenin/Wnt 途径的一部分，要么是相互的 受其监管。在本提案中，我们将专门研究 ß-catenin 在病理生理学中的作用 婴儿痉挛症。我们将通过删除 APC 来检查增加 ß-catenin 的效果，并且独立于 APC，关于痉挛行为、癫痫发作和脑电图活动。接下来，我们将认真执行 操作β-catenin的药代动力学、药效学和不良反应分析 发展。最后，我们将确定将 β-连环蛋白水平恢复到正常水平是否可以减轻痉挛和癫痫发作 在以后的生活中。该提案将解决 ß-连环蛋白在痉挛病理生理学中的作用，提供新的方法 用于临床前分析的小鼠模型，并为该疾病引入大量新的潜在治疗靶点 IS的治疗。